Review ArticleLactoferrin A Modulator for Immunity againstTuberculosis Related Granulomatous Pathology

Jeffrey K Actor12

1UTHealth Department of Pathology University of Texas-Houston Medical School Houston TX 77030 USA2Program in Immunology Graduate School of Biomedical Sciences Houston TX 77030 USA

Correspondence should be addressed to Jeffrey K Actor jeffreykactoruthtmcedu

Received 8 October 2015 Accepted 30 November 2015

Academic Editor Magdalena Klink

Copyright copy 2015 Jeffrey K Actor This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

There is great need for a therapeutic that would limit tuberculosis related pathology and thus curtail spread of disease betweenindividuals by establishing a ldquofirebreakrdquo to slow transmission A promising avenue to increase current therapeutic efficacy maybe through incorporation of adjunct components that slow or stop development of aggressive destructive pulmonary pathologyLactoferrin an iron-binding glycoprotein found in mucosal secretions and granules of neutrophils is just such a potential adjuncttherapeutic agent The focus of this review is to explore the utility of lactoferrin to serve as a therapeutic tool to investigateldquodisruptionrdquo of the mycobacterial granuloma Proposed concepts for mechanisms underlying lactoferrin efficacy to controlimmunopathology are supported by data generated based on in vivomodels using nonpathogenic trehalose 661015840-dimycolate (TDMcord factor)

1 Introduction

There is great need for a therapeutic that would limit tubercu-losis pathology and thus limit spread of disease between indi-viduals by establishment of a ldquofirebreakrdquo to slow transmission[1] A promising avenue to increase current therapeutic effi-cacy may be through incorporation of adjunct componentsthat curtail development of aggressive destructive pulmonarypathology Lactoferrin an iron-binding glycoprotein foundin mucosal secretions and granules of neutrophils is justsuch a potential adjunct therapeutic agent The focus ofthis review is therefore to explore the utility of lactoferrinto serve as a therapeutic tool to investigate ldquodisruptionrdquo ofthe mycobacterial granuloma during infection In additionmechanisms for lactoferrin efficacy will be proposed basedon manipulation of trehalose 661015840-dimycolate (TDM cordfactor) immunopathology developed in a nonpathogenic invivo setting The studies here indicate that lactoferrin may bea useful adjunct therapeutic for amelioration of pathologicalresponse to mycobacteria

Mycobacterial Granulomas Primary Tuberculosis (TB)begins with infection that spreads via lymphatics and blood

stream before inducing systemic immunity that contains andcontrols the organisms within granulomas Induction of theMycobacterium tuberculosis (MTB) disease-induced granu-loma is dependent on factors involved in response initiationand associated immune activity [2ndash7] In the most extremesimplification of the process the underlying initiator is theexistence of a poorly degradable persistent antigenic sourceMycobacterial organisms survive within macrophagesexhibiting slow release of antigens to recruited cells The netresult of granuloma formation is of initial benefit to boththe host and the organism allowing control of infectionwhile providing a place for organisms to hide until time forexpansion and subsequent transmission to other individuals[8 9] In reality the organism exhibits effects on tissuessurrounding infected cells through released potent bioactivecell wall constituents [10] New paradigms implicate inflam-matory processes beginning with induction of a lipid-relatednecrotic pneumonia that may transition to become the focusof granulomas and fibrocaseous disease [11ndash14] Regardlessof event initiation it is clear that over time the disruptivenature of the granuloma causes harm to the infected host

Our laboratory has studied granuloma pathobiology indetail in mouse models of tuberculosis [8 15ndash20] and made

Hindawi Publishing CorporationMediators of InflammationVolume 2015 Article ID 409596 10 pageshttpdxdoiorg1011552015409596

2 Mediators of Inflammation

contributions to factors involved in the development of pro-gressive pathology Furthermore we utilized our knowledgeto design protocols to maximize immune-modulating agentsto investigate the relationship of protection and pathology[21ndash24] An area of study used lactoferrin as an immunemod-ulator to augment the BCG vaccine and relative responsesto mycobacterial antigens [22ndash27] leading to reduction inpathological damage after challenge with virulent organismsThey were the observations made during development ofhuman lactoferrin as a vaccine adjuvant that led us tobelieve it also had utility as an immune modulator to reduceinflammation during granuloma formation [23 28]

Many of the molecular mechanisms involved in develop-ment maintenance and initiation of factors related to myco-bacterial pathology in the lung have been identified A systemof choice is amodel based on the noninfectious proinflamma-tory granuloma induced by the major surface glycolipid frompathogenic organisms developed by multiple investigatorsincluding Retzinger Behling and Hunter [29ndash32] This haspermitted a global understanding of the immunopathologyInjection with cord factor (trehalose 661015840-dimycolate TDM)induces transient pulmonary granulomas that increase innumber and complexity over a 7-day period followed byresolution of pathology [33] Manipulation of host factors hasallowed definition of immune parameters contributing to theinitiation maintenance and resolution phases of response

TheCord Factor Model of theMycobacterial Granuloma Cordfactor (trehalose 661015840-dimycolate TDM) is the most abun-dant glycolipid produced on the surface of mycobacterialorganisms TDM plays multiple roles in the pathogenesis ofMTB [34 35] including the formation of caseation in thelung after infection [36] TDM formulated into an emulsionor placed on beads and injected intravenously into miceinduces a lung pathology that mimics many aspects ofearly MTB infection including granulomatous response andthe production of proinflammatory cytokines The TDMmodel of granuloma formation has been used to elucidatethe immunological factors involved in the granulomatousresponse [37 38] Furthermore TDM can induce activatedforeign body type granulomas in naıve mice [33 39] andhypersensitivity (immune) granulomas in appropriately sen-sitized mice [36 40 41] Therefore this model system isideal for exploring the potential of immunomodulators toalter granuloma structure with perhaps specific utility toextrapolate findings to immune related pathology identifiedduring clinical manifestation of tuberculosis disease

The interactions of mycobacteria and its lipid com-ponents with cell surface antigens to elicit inflammatoryresponses have been characterized [42] and are the subject ofother papers and reviews [43ndash46] Recent discoveries link theC-type lectin mincle [47ndash49] as a prime candidate receptorfor TDM This is augmented in part by interactions withMARCO TLR2 andor CD14 all of which are critical tomediate activity [44] These are combined with internal-ized signaling events that possibly function through Card9-dependentmechanisms [50 51] Suffice to say TDMrsquos interac-tionwith surface ldquoreceptorsrdquo is critical for initiation of cellularactivation of TLR and NOD pathways as well as control of

intracellular trafficking events [52 53] Using knockout miceestablished patterns of cytokine production were found tobe associated with pathology This allowed the delineationof the major molecular events in innate establishment andmaintenance of the TDM induced granuloma [17 33 37 3841 52ndash56] A link was also established to adaptive (T-cellmediated) hypersensitive responses critical for developmentof pathological granulomas [57 58] (Figure 1)

Lactoferrin as a First-Line Defense Protein Lactoferrin is amonomeric 80 kDa single polypeptide chain contained inmost mammalian exocrine secretions including milk tearssaliva and bronchial and intestinal secretions It is alsopresent in the secondary granules of neutrophils It is consid-ered a first-line defense protein involved in protection againstmicrobial infections [59 60] and subsequent development ofsystemic inflammatory response syndrome (SIRS) and sepsis[61ndash67] More recently lactoferrin has been implicated inimmunoregulatory functions [62 68ndash73] and modulation ofvaccine function [22ndash27]

There are two primary forms of human lactoferrinone contained in exocrine secretions and the other presentin secondary granules of neutrophils The two forms areidentical in their amino acid sequence but differ in glycancontent [75 76] While the secreted form is thought tobe involved in the host defense against microbial infec-tion at mucosal sites granulocyticneutrophilic lactoferrinhas notable immunomodulatory function [74] Neutrophiliclactoferrin is an integral part of the cytokine-mediatedcascade during insult-induced metabolic imbalance [77ndash81](Figure 2) with a subset of biological properties uniquefrom lactoferrins produced at other sources by other cellphenotypes [82]

The key to understanding the molecular basis of itsactivities is thought to reside in both patterns of glycosylationand sialylation [83 84] The primary structure of humanLF is characterized by a single polypeptide chain containing692 amino acids organized in two highly homologous lobesdesignated the N- and C-lobe each capable of bindingsingle ferric ion (Fe+++) Lactoferrin is a glycoprotein andin humans the glycans are the N-acetyllactosaminic type12057213-fucosylated on the N-acetyl-glucosamine residue linkedto the peptide chain There are three possible N-linkedglycosylation sites in hLF one at Asn138 a second site atAsn479 and a third site at Asn624 differential utilizationof these sites results in distinct glycosylation variants [85]Many observed activities of LF are dependent upon specificglycosylation patterns For example the immunoregulatoryactivity of LF in humans is dependent on the interaction ofthis glycoprotein with a receptor specific for sialic acid anddirect lymphocyte activation by LF requires sialylation [86]

2 Lactoferrin as an Immune Modulator

21 Lactoferrin A Balance between Potentiation and Media-tion Lactoferrin bridges innate and adaptive immune func-tions by regulating specific target cell responses [87 88] Byacting as a homeostatic modulator lactoferrin can work bothends of the immunological spectrum to increase low response

Mediators of Inflammation 3

TDM

Macrophage

TDM

TDM ComplementIL-6

Initiation phase Maintenance phase

IL-6

Active cortisol Inactive cortisol

Destructive pathology

Hypersensitive regulation

T-cell

Innate Adaptive

11120573HSDI 11120573HSDII

TNF-120572 (C5a)

Figure 1 Mechanisms of TDM-induced granulomas Cytokines and immune-regulatory factors induced by mycobacterial glycolipidTDM play key roles in the development of pathology induced during mycobacterial infection Model systems using isolated TDM allowidentification of host immune function to extrapolate findings of immune related pathology occurring during clinical manifestation oftuberculosis disease [17 33 37 38 41 52ndash58]

Insult RepairAcute inflammation

NeutrophilDendritic cellMacrophage

LFLF

LFLF

Augmentlow antigenic response

Modulateaggressive inflammation

Proinflammatory(TNF120572 IL-6

IL-1120573 and IL-8)

Figure 2 Lactoferrin mediation of insult-induced inflammation Insult defined as infection trauma or chronic illness leads to activation ofmacrophage and dendritic cells Activated neutrophils degranulate at the site of injury and releasemassive amounts of lactoferrin Lactoferrinin turn can either augment low level responses or modulate overaggressive cytokine activity Both effects serve to control inflammation toassist with tissue repair after insult (modified from [74])

or dampen aggressive ones It has direct capacity to regulateboth proinflammatory and anti-inflammatory responses [6263] The utility of such an immune mediator represents anovel therapeutic agent dependent on elicited responses foroutcome [74] A direct example of how lactoferrin canmanip-ulate outcomes of response is exemplified by knowledge thatlactoferrin can bind to the soluble CD14 and the CD14LPScomplex [89] an interaction which has been shown to medi-ate toll-like receptor-4 (TLR-4) pattern recognition events[90] Indeed lactoferrin mediated events related to TLR-4may represent a strong strategy to block excessive antigenpresenting cell activation [91]What remains unknown is howlactoferrin might affect PAMP signaling processes duringtuberculosis infection While some data suggests that Myd88pathways may not be as critical in transition to adaptivedefense against tuberculosis [92] more recent studies suggestthat subversion of theTLR-2-MyD88 pathway is an important

factor in intracellular processing [93] In either case it wouldseem that lactoferrin may function as a regulator in anindependent manner during immune modulation during TBchallenge

We and collaborators investigated lactoferrin to modifyinnate events during inflammation such as those seen dur-ing systemic inflammatory response syndrome and modelsof oxidative stress [78 94ndash97] Our laboratory also spentyears investigating lactoferrin augmentation of the BCGvaccine to protect against subsequent challenge with virulentMTB functioning through strong induction of cell-mediatedimmunity [64 98] and generation of IFN-120574 antigen specificrecall responses [24 26 27 69 98 99] Lactoferrin has aprofound modulatory action on adaptive immune functions[70 73] by promoting maturation of T-cell precursors intocompetent helper cells and differentiation of immature B-cells into efficient antigen presenting cells [100] The innate

4 Mediators of Inflammation

and adaptive events are not mutually exclusive but rathercomplementary and lend credence to utility of lactoferrin forimmune homeostasis in insult-inducedmetabolic disparities

22 Lactoferrin Modulation of the Tuberculoid GranulomaAn examination of lactoferrin to modulate granulomatousresponses was performed in mice injected intravenouslywith TDM A subset of mice were intravenously given 1mgbovine lactoferrin 24 h after TDM challenge Lung tissue wasanalyzed for histological response and for the productionof proinflammatory mediators Treatment with lactoferrinshowed statistically significant fewer and smaller granulomascompared to TDM alone [101] Of importance the protectiveproinflammatory mediators were not statistically diminishedby lactoferrin treatment Nontreated mice demonstrateda granuloma formation that correlated with an increasedproduction of tumor necrosis factor-120572 (TNF-120572) interleukin-(IL-) 1120573 and IL-6This is important as it suggests that innatefunctions critical during infectious disease are maintainedeven though immunopathology is reduced by treatment

In a similar manner the utility of oral delivered lactofer-rin was also examined As previously mentioned mice givenTDM alone also showed marked and statistically significantincreased production of proinflammatory cytokines TNF-120572IL-6 IL-1120573 IL-12p40 and KC (keratinocyte chemoattrac-tant) Oral treatment with recombinant human LF (producedin CHO cells) at 1 or 2mg led to statistically significantreduction (119901 lt 005) in cytokine levels at 7 days after TDMadministration in BALBc and C57BL6 mice (Figure 3 onlyBALBc data shown) This is consistent with reports thatthese proinflammatory cytokines are protective in similarinfectious challenge models [102 103] while molecules suchas IL-10 which are limited after lactoferrin administrationare detrimental during infection [104] In a similar mannerbovine lactoferrin given orally at 1 or 2mg doses also showedstatistical reduction in most cytokines although there wasgreater consistency of response in BALBc mice than thatfor the C57BL6 strain In particular the 1mg dosage ofrecombinant LF showed the most consistency in its ability tocontrol (reduce) production of proinflammatory cytokines

Of major interest the oral delivered human recombinantlactoferrin demonstrated biologically relevant reduction inpathology (Figure 4) similar to that seen in the publishedresults using intravenous administered bovine lactoferrin Infact both the bovine and the recombinant human lactoferrinswere able to reduce inflammation due toTDMchallenge withdoses of 1 or 2mg given on days 4 and 6 after challenge result-ing in clear reduction in granuloma size and frequency Themost effective dose for the recombinant human lactoferrinwas seen when used at 1mg with some efficacy also seengiven as a single dose only on day 4 (data not shown)

Another point of special interest is that the classicalcytokines critical for control of MTB namely TNF-120572 IL-6 and IFN-120574 were not statistically altered by lactoferrintreatment [105] This gave confidence that treatment wouldretain control of organisms after infectious challenge Indeedthis is what was seen in a similar experiment performedusing bovine lactoferrin administered in drinking water(5mgmL) given to mice aerosol infected with virulent

mycobacteria (Erdman strain) [105] While bacterial loadin tissue was slightly reduced the major change was ame-lioration of granulomatous severity It is noteworthy thatbovine lactoferrin reduced bacterial burden accompaniedby an increase in classical proinflammatory responses whiledecreasing overall lung immunopathology [105] Specificallylactoferrin-treated mice increased numbers of CD4+ IFN-120574+ and IL-17 producing cells in the lung It was shown thatlactoferrin by itself was not bactericidal but rather enhancedIFN-120574mediatedMTBkilling bymacrophages in a nitric oxidedependent manner These studies indicate that lactoferrinmay be a novel therapeutic for the treatment of tuberculosisand may be useful in infectious diseases to reduced immune-mediated tissue damage

Finally the molecular mechanism underlying resolutionof the granulomatous lesion is not yet defined However arole for maintaining strong Th1 responses could possibly beattributed to relationships due to COX-2 inhibition by lacto-ferrin in inducedmacrophages (WBCs) [106] and knowledgethat prostaglandin E2 (PGE2) is able to shift T helperresponses during BCG activation of innate macrophages[107] It had been previously demonstrated that the PGE2response could be reversed by a COX-2 inhibitor [108]Therefore lactoferrin functioning as a COX-2 inhibitor couldbe an indirect method to maintain proinflammatory activityof macrophages when infected

3 Conclusions

The studies here indicate that lactoferrin may be a usefuladjunct therapeutic for amelioration of pathological responsein an in vivo model of TB granulomas Of high importancethe change in pathology is accomplished with no loss ofinnate immune function which would be critical for defenseagainst pathogenic organisms A question arises concerningthe utility of using heterologous lactoferrins in the mousemodel Preliminary investigations show that a newly devel-oped recombinant mouse lactoferrin produced in CHO sta-ble cell lines functions in a manner very similar to the bovineand human recombinant to limit immunopathology afteroral delivery Mechanistically it was shown that the mouseand human forms are nearly identical in modulation of den-dritic cell function in response to Bacillus Calmette-Guerin(BCG) [25 109] The dendritic cell population respondswith increased inflammatory cytokines and a shift towardsMHC Class II expression when BCG is combined withthe lactoferrins However more recent investigations revealthat the macrophage population has a unique trend withpotentially decreased antigen presentation and subsequentT-cell stimulatory activity [110] Overall these results givegreat confidence to move forward to test novel recombinantlactoferrins for adjunct clinical therapeutic effects to alterimmunedependent granulomatous pathology occurring dur-ing tuberculosis infection in humansThe studies here woulddrive the next goals to explore novel recombinant lactoferrinsto mediate reductions in lung histopathology and bacterialburden in BSL3 in vivo models of MTB infection Further-more we hypothesize that reduction of TB-induced inflam-matory pathology would also allow enhanced penetration

Mediators of Inflammation 5

2000

1500

1000

500

0

TDM

Treatment

lowast lowast lowast lowast

IL-12

p40

conc

entr

atio

n (p

glu

ng)

Lung IL-12p40

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

6000

2000

3000

4000

5000

1000

0

TDM

Treatment

lowast lowast lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

Lung TNF-120572

IL-6

conc

entr

atio

n (p

glu

ng)

2000

1500

1000

500

0

TDM

Treatment

lowast lowast lowast

Lung IL-6

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

40000

20000

30000

10000

0

TDM

Treatment

lowast lowast lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

IL-1120573

conc

entr

atio

n (p

glu

ng)

Lung IL-1120573

IL-10

conc

entr

atio

n (p

glu

ng)

Lung IL-10300

200

100

0

TDM

Treatment

lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

TNF-120572

conc

entr

atio

n (p

glu

ng)

Figure 3 Proinflammatory cytokines in lungs are reduced in lactoferrin-treated mice after challenge with TDM BALBc mice werechallenged with 25120583g TDM in a water-in-oil emulsion Bovine or recombinant human lactoferrin (1 or 2mg) was orally administered at4 and 6 days after TDM challenge Methods of TDM induction are described elsewhere [38 101] Lungs were isolated on day 7 after TDMchallenge cytokines were assessed by ELISA values are shown per lung for individual mice lowast119901 lt 005 compared to the TDM emulsion alonetreated mice

of antimycobacterial drugs to sites harboring infectiousagents in essence permitting an efficient mechanism totreat TB infection Overall these studies permit confidenceto target clinical implications and test novel strategies to

modulate MTB-induced granulomatous responses to allowmore efficacious delivery of therapeutics to mycobacterialorganisms residing within lung tissue during activemycobac-terial disease states

6 Mediators of Inflammation

BALB

cC5

7BL

6TDM alone TDM + LFNaiumlve

TDM alone TDM + LFNaiumlve

Figure 4 Granuloma response to TDM challenge in oral recombinant human lactoferrin-treated mice Two strains of mice (BALBc andC57BL6) were challenged with 25120583g TDM in a water-in-oil emulsion as described [38 101] Human lactoferrin (recombinant 1mg) wasgiven orally at 4 and 6 days after TDM challenge Comparisons are made to naıve mice and to mice only administered TDM HampE staining100x Representative sections from mice at 7 days after TDM challenge119873 ge 8 per experimental group

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

This work was supported in part by NIH Grants 1R41-AI117990-01 and R42-AI051050-05The authors acknowledgePharmaReview Corp (Houston TX) for their kind gifts oflactoferrins and thank Marian L Kruzel PhD and Shen-An Hwang PhD for their insights into the research projectThe authors also thank Ron Gadot and Ai Yamazaki for theirtechnical contributions to the research presented

References

[1] P E M Fine ldquoHerd immunity history theory practicerdquo Epide-miologic Reviews vol 15 no 2 pp 265ndash302 1993

[2] L Ramakrishnan ldquoRevisiting the role of the granuloma intuberculosisrdquo Nature Reviews Immunology vol 12 no 5 pp352ndash366 2012

[3] M Silva Miranda A Breiman S Allain F Deknuydt andF Altare ldquoThe tuberculous granuloma an unsuccessful hostdefence mechanism providing a safety shelter for the bacteriardquoClinical amp Developmental Immunology vol 2012 Article ID139127 14 pages 2012

[4] S A Khader J Rangel-Moreno J J Fountain et al ldquoIn amurinetuberculosis model the absence of homeostatic chemokinesdelays granuloma formation and protective immunityrdquo Journalof Immunology vol 183 no 12 pp 8004ndash8014 2009

[5] AMCooper and S A Khader ldquoThe role of cytokines in the ini-tiation expansion and control of cellular immunity to tubercu-losisrdquo Immunological Reviews vol 226 no 1 pp 191ndash204 2008

[6] IMOrme and R J Basaraba ldquoThe formation of the granulomain tuberculosis infectionrdquo Seminars in Immunology vol 26 no6 pp 601ndash609 2014

[7] I M Orme R T Robinson and A M Cooper ldquoThe balancebetween protective and pathogenic immune responses in theTB-infected lungrdquo Nature Immunology vol 16 no 1 pp 57ndash632014

[8] J K Actor M Olsen C Jagannath and R L Hunter ldquoRela-tionship of survival organism containment and granulomaformation in acute murine tuberculosisrdquo Journal of Interferonand Cytokine Research vol 19 no 10 pp 1183ndash1193 1999

[9] J L Flynn and J Chan ldquoWhatrsquos good for the host is good for thebugrdquo Trends in Microbiology vol 13 no 3 pp 98ndash102 2005

[10] D G Russell ldquoMycobacterium tuberculosis and the intimatediscourse of a chronic infectionrdquo Immunological Reviews vol240 no 1 pp 252ndash268 2011

[11] R L Hunter ldquoPathology of post primary tuberculosis of thelung an illustrated critical reviewrdquo Tuberculosis vol 91 no 6pp 497ndash509 2011

[12] R L Hunter J K Actor S-A Hwang V Karev and C Jagan-nath ldquoPathogenesis of post primary tuberculosis immunity andhypersensitivity in the development of cavitiesrdquoAnnals of Clini-cal and Laboratory Science vol 44 no 4 pp 365ndash387 2014

[13] K J Welsh R L Hunter and J K Actor ldquoTrehalose 661015840-dimy-colatemdasha coat to regulate tuberculosis immunopathogenesisrdquoTuberculosis vol 93 supplement pp S3ndashS9 2013

[14] M-J Kim H C Wainwright M Locketz et al ldquoCaseation ofhuman tuberculosis granulomas correlates with elevated hostlipid metabolismrdquo EMBO Molecular Medicine vol 2 no 7 pp258ndash274 2010

[15] J K Actor E Breij R A Wetsel H Hoffmann R L HunterJr and C Jagannath ldquoA role for complement C5 in organ-ism containment and granulomatous response during murine

Mediators of Inflammation 7

tuberculosisrdquo Scandinavian Journal of Immunology vol 53 no5 pp 464ndash474 2001

[16] J K Actor C D Leonard V E Watson et al ldquoCytokinemRNAexpression and serumcortisol evaluation duringmurinelung inflammation induced by Mycobacterium tuberculosisrdquoCombinatorial Chemistry and High Throughput Screening vol3 no 4 pp 343ndash351 2000

[17] C W Borders A Courtney K Ronen et al ldquoRequisite role forcomplement C5 and the C5a receptor in granulomatous res-ponse to mycobacterial glycolipid trehalose 661015840-dimycolaterdquoScandinavian Journal of Immunology vol 62 no 2 pp 123ndash1302005

[18] R L Hunter C Jagannath and J K Actor ldquoPathology of post-primary tuberculosis in humans and mice contradiction oflong-held beliefsrdquo Tuberculosis vol 87 no 4 pp 267ndash278 2007

[19] C Jagannath H Hoffmann E Sepulveda J K Actor R AWetsel and R L Hunter Jr ldquoHypersusceptibility of AJ miceto tuberculosis is in part due to a deficiency of the fifth comple-ment component (C5)rdquo Scandinavian Journal of Immunologyvol 52 no 4 pp 369ndash379 2000

[20] V E Watson L L Hill L B Owen-Schaub et al ldquoApoptosis inMycobacterium tuberculosis infection in mice exhibiting variedimmunopathologyrdquo Journal of Pathology vol 190 no 2 pp 211ndash220 2000

[21] R H Copenhaver E Sepulveda L Y Armitige et al ldquoAmutantof Mycobacterium tuberculosis H37Rv that lacks expressionof antigen 85A is attenuated in mice but retains vaccinogenicpotentialrdquo Infection and Immunity vol 72 no 12 pp 7084ndash7095 2004

[22] S-A Hwang R Arora M L Kruzel and J K Actor ldquoLactofer-rin enhances efficacy of the BCG vaccine comparison betweentwo inbred mice strains (C57BL6 and BALBc)rdquo Tuberculosisvol 89 supplement 1 pp S49ndashS54 2009

[23] S-A Hwang K Wilk M L Kruzel and J K Actor ldquoA novelrecombinant human lactoferrin augments the BCG vaccine andprotects alveolar integrity upon infection with Mycobacteriumtuberculosis in micerdquo Vaccine vol 27 no 23 pp 3026ndash30342009

[24] S-A Hwang K M Wilk M Budnicka et al ldquoLactoferrinenhanced efficacy of the BCG vaccine to generate host protec-tive responses against challenge with virulent Mycobacteriumtuberculosisrdquo Vaccine vol 25 no 37-38 pp 6730ndash6743 2007

[25] S-A Hwang and J K Actor ldquoLactoferrin modulation of BCG-infected dendritic cell functionsrdquo International Immunologyvol 21 no 10 pp 1185ndash1197 2009

[26] S-A Hwang M L Kruzel and J K Actor ldquoLactoferrin aug-ments BCG vaccine efficacy to generate T helper response andsubsequent protection against challenge with virulentMycobac-terium tuberculosisrdquo International Immunopharmacology vol 5no 3 pp 591ndash599 2005

[27] K M Wilk S-A Hwang and J K Actor ldquoLactoferrin mod-ulation of antigen-presenting-cell response to BCG infectionrdquoPostepy Higieny i Medycyny Doswiadczalnej vol 61 pp 277ndash282 2007

[28] S A Hwang M L Kruzel and J K Actor ldquoCHO expressedrecombinant human lactoferrin as an adjuvant for BCGrdquoInternational Journal of Immunopathology and Pharmacologyvol 28 no 4 pp 452ndash468 2015

[29] R L Perez J Roman G W Staton Jr and R L HunterldquoExtravascular coagulation and fibrinolysis in murine lunginflammation induced by the mycobacterial cord factor

trehalose-66rsquo-dimycolaterdquoAmerican Journal of Respiratory andCritical Care Medicine vol 149 no 2 pp 510ndash518 1994

[30] C A Behling R L Perez M R Kidd G W Staton Jr and RL Hunter ldquoInduction of pulmonary granulomas macrophageprocoagulant activity and tumor necrosis factor-alpha by tre-halose glycolipidsrdquo Annals of Clinical and Laboratory Sciencevol 23 no 4 pp 256ndash266 1993

[31] G S Retzinger S C Meredith R L Hunter K Takayama andF J Kezdy ldquoIdentification of the physiologically active state ofthe mycobacterial glycolipid trehalose 661015840-dimycolate and therole of fibrinogen in the biologic activities of trehalose 661015840-dimycolate monolayersrdquo Journal of Immunology vol 129 no 2pp 735ndash744 1982

[32] G S Retzinger S C Meredith K Takayama R L Hunter andF J Kezdy ldquoThe role of surface in the biological activities oftrehalose 661015840-dimycolate Surface properties and developmentof amodel systemrdquoThe Journal of Biological Chemistry vol 256no 15 pp 8208ndash8216 1981

[33] R L Perez J Roman S Roser et al ldquoCytokinemessage and pro-tein expression during lung granuloma formation and resolu-tion induced by the mycobacterial cord factor trehalose-661015840-dimycolaterdquo Journal of Interferon amp Cytokine Research vol 20no 9 pp 795ndash804 2000

[34] R L Hunter M R Olsen C Jagannath and J K Actor ldquoMult-iple roles of cord factor in the pathogenesis of primary sec-ondary and cavitary tuberculosis including a revised descrip-tion of the pathology of secondary diseaserdquo Annals of Clinicaland Laboratory Science vol 36 no 4 pp 371ndash386 2006

[35] J K Actor ldquoMycobacterial TDM a coat to modulate post pri-mary pathogenesisrdquoMycobacterial Diseases vol 2 no 1 articlee106 2012

[36] R L HunterMOlsen C Jagannath and J K Actor ldquoTrehalose661015840-dimycolate and lipid in the pathogenesis of caseatinggranulomas of tuberculosis in micerdquo The American Journal ofPathology vol 168 no 4 pp 1249ndash1261 2006

[37] A N Abbott T V Guidry K J Welsh et al ldquo11Beta-hydroxy-steroid dehydrogenases are regulated during the pulmonarygranulomatous response to the mycobacterial glycolipidtrehalose-661015840-dimycolaterdquo NeuroImmunoModulation vol 16no 3 pp 147ndash154 2009

[38] K J Welsh A N Abbott S-A Hwang et al ldquoA role for tumournecrosis factor-120572 complement C5 and interleukin-6 in theinitiation and development of the mycobacterial cord factortrehalose 661015840-dimycolate induced granulomatous responserdquoMicrobiology vol 154 no 6 pp 1813ndash1824 2008

[39] A Bekierkunst and E Yarkoni ldquoGranulomatous hypersensitiv-ity to trehalose-66rsquo-dimycolate (cord factor) in mice infectedwith BCGrdquo Infection and Immunity vol 7 no 4 pp 631ndash6381973

[40] H Yamagami TMatsumoto N Fujiwara et al ldquoTrehalose 661015840-dimycolate (cord factor) ofMycobacterium tuberculosis inducesforeign-body- and hypersensitivity-type granulomas in micerdquoInfection and Immunity vol 69 no 2 pp 810ndash815 2001

[41] T VGuidry R LHunter Jr and J K Actor ldquoMycobacterial gly-colipid trehalose 661015840-dimycolate-induced hypersensitive gran-ulomas contribution of CD4+ lymphocytesrdquoMicrobiology vol153 no 10 pp 3360ndash3369 2007

[42] E Rhoades F-F Hsu J B Torrelles J Turk D Chatterjee andD G Russell ldquoIdentification and macrophage-activating activ-ity of glycolipids released from intracellular Mycobacteriumbovis BCGrdquoMolecular Microbiology vol 48 no 4 pp 875ndash8882003

8 Mediators of Inflammation

[43] MMHossain andM-N Norazmi ldquoPattern recognition recep-tors and cytokines in Mycobacterium tuberculosis infectionmdashthe double-edged swordrdquo BioMed Research International vol2013 Article ID 179174 18 pages 2013

[44] D M E Bowdish K Sakamoto M-J Kim et al ldquoMARCOTLR2 and CD14 are required for macrophage cytokine res-ponses to mycobacterial trehalose dimycolate and Mycobac-terium tuberculosisrdquo PLoS Pathogens vol 5 no 6 Article IDe1000474 2009

[45] E Mortaz I M Adcock P Tabarsi et al ldquoInteraction of patternrecognition receptors withMycobacterium tuberculosisrdquo Journalof Clinical Immunology vol 35 no 1 pp 1ndash10 2015

[46] D G Russell ldquoWho puts the tubercle in tuberculosisrdquo NatureReviews Microbiology vol 5 no 1 pp 39ndash47 2007

[47] H Schoenen B Bodendorfer K Hitchens et al ldquoCutting Edgemincle is essential for recognition and adjuvanticity of themycobacterial cord factor and its synthetic analog trehalose-dibehenaterdquo Journal of Immunology vol 184 no 6 pp 2756ndash2760 2010

[48] E Ishikawa T Ishikawa Y S Morita et al ldquoDirect recognitionof themycobacterial glycolipid trehalose dimycolate by C-typelectinMinclerdquo Journal of ExperimentalMedicine vol 206 no 13pp 2879ndash2888 2009

[49] I Matsunaga and D B Moody ldquoMincle is a long soughtreceptor for mycobacterial cord factorrdquo Journal of ExperimentalMedicine vol 206 no 13 pp 2865ndash2868 2009

[50] K Werninghaus A Babiak O Gross et al ldquoAdjuvanticity ofa synthetic cord factor analogue for subunit Mycobacteriumtuberculosis vaccination requires FclR120574-Syk-Card9-dependentinnate immune activationrdquo Journal of Experimental Medicinevol 206 no 1 pp 89ndash97 2009

[51] S LeibundGut-LandmannOGrossM J Robinson et al ldquoSyk-and CARD9-dependent coupling of innate immunity to theinduction of T helper cells that produce interleukin 17rdquo NatureImmunology vol 8 no 6 pp 630ndash638 2007

[52] J Indrigo R L Hunter Jr and J K Actor ldquoInfluence of tre-halose 661015840-dimycolate (TDM) during mycobacterial infectionof bone marrow macrophagesrdquoMicrobiology vol 148 no 7 pp1991ndash1998 2002

[53] J Indrigo R L Hunter Jr and J K Actor ldquoCord factortrehalose 661015840-dimycolate (TDM) mediates trafficking eventsduringmycobacterial infection ofmurinemacrophagesrdquoMicro-biology vol 149 no 8 pp 2049ndash2059 2003

[54] J K Actor M Olsen R L Hunter Jr and Y-J Geng ldquoDys-regulated response to mycobacterial cord factor trehalose-661015840-dimycolate inCD1Dminusminusmicerdquo Journal of Interferon andCytokineResearch vol 21 no 12 pp 1089ndash1096 2001

[55] J K Actor J Indrigo C M Beachdel et al ldquoMycobacterial gly-colipid cord factor trehalose 661015840-dimycolate causes a decreasein serum cortisol during the granulomatous responserdquo Neuro-ImmunoModulation vol 10 no 5 pp 270ndash282 2002

[56] A N Abbott K J Welsh S-A Hwang et al ldquoIL-6 mediates11120573hsd type 2 to effect progression of the mycobacterial cordfactor trehalose 661015840-dimycolate-induced granulomatous res-ponserdquo NeuroImmunoModulation vol 18 no 4 pp 212ndash2252011

[57] T V Guidry M Olsen K-S Kil R L Hunter Jr Y-J Gengand J K Actor ldquoFailure of CD1Dminusminusmice to elicit hypersensitivegranulomas to mycobacterial cord factor trehalose 661015840-dimy-colaterdquo Journal of Interferon amp Cytokine Research vol 24 no 6pp 362ndash371 2004

[58] TVGuidry R LHunter Jr and J KActor ldquoCD3+ cells transferthe hypersensitive granulomatous response to mycobacterialglycolipid trehalose 661015840-dimycolate in micerdquoMicrobiology vol152 no 12 pp 3765ndash3775 2006

[59] B Lonnerdal and S Iyer ldquoLactoferrin molecular structure andbiological functionrdquoAnnual Review of Nutrition vol 15 pp 93ndash110 1995

[60] L Sanchez M Calvo and J H Brock ldquoBiological role of lacto-ferrinrdquo Archives of Disease in Childhood vol 67 no 5 pp 657ndash661 1992

[61] P Manzoni M Rinaldi S Cattani et al ldquoBovine lactoferrinsupplementation for prevention of late-onset sepsis in verylow-birth-weight neonates a randomized trialrdquo Journal of theAmerican Medical Association vol 302 no 13 pp 1421ndash14282009

[62] S Baveye E Elass J Mazurier G Spik and D Legrand ldquoLacto-ferrin a multifunctional glycoprotein involved in the modula-tion of the inflammatory processrdquoClinical Chemistry and Labo-ratory Medicine vol 37 no 3 pp 281ndash286 1999

[63] R D Baynes and W R Bezwoda ldquoLactoferrin and the inflam-matory responserdquo Advances in Experimental Medicine andBiology vol 357 pp 133ndash141 1994

[64] M Kocieba M Zimecki M Kruzel and J Actor ldquoThe adjuvantactivity of lactoferrin in the generation of DTH to ovalbumincan be inhibited by bovine serum albumin bearing alpha-D-mannopyranosyl residuesrdquoCellularampMolecular Biology Lettersvol 7 no 4 pp 1131ndash1136 2002

[65] C G Turin A Zea-Vera A Pezo et al ldquoLactoferrin for pre-vention of neonatal sepsisrdquo Biometals vol 27 no 5 pp 1007ndash1016 2014

[66] T Siqueiros-Cendon S Arevalo-Gallegos B F Iglesias-Figueroa I A Garcıa-Montoya J Salazar-Martınez and QRascon-Cruz ldquoImmunomodulatory effects of lactoferrinrdquo ActaPharmacologica Sinica vol 35 no 5 pp 557ndash566 2014

[67] D Latorre F Berlutti P Valenti S Gessani and P Puddu ldquoLFimmunomodulatory strategies mastering bacterial endotoxinrdquoBiochemistry and Cell Biology vol 90 no 3 pp 269ndash278 2012

[68] D Legrand ldquoLactoferrin a key molecule in immune andinflammatory processesrdquo Biochemistry and Cell Biology vol 90no 3 pp 252ndash268 2012

[69] S-A Hwang K M Wilk Y A Bangale M L Kruzel and JK Actor ldquoLactoferrin modulation of IL-12 and IL-10 responsefrom activated murine leukocytesrdquo Medical Microbiology andImmunology vol 196 no 3 pp 171ndash180 2007

[70] M Zimecki J Mazurier G Spik and J A Kapp ldquoHuman lacto-ferrin induces phenotypic and functional changes in murinesplenic B cellsrdquo Immunology vol 86 no 1 pp 122ndash127 1995

[71] M Zimecki J Artym G Chodaczek et al ldquoImmunoregulatoryfunction of lactoferrin in immunosuppressed and autoimmuneanimalsrdquo Postepy Higieny i Medycyny Doswiadczalnej vol 61pp 283ndash287 2007

[72] M Zimecki and M L Kruzel ldquoMilk-derived proteins andpeptides of potential therapeutic and nutritive valuerdquo Journal ofExperimental Therapeutics and Oncology vol 6 no 2 pp 89ndash106 2007

[73] M Zimecki RMiędzybrodzki J Mazurier andG Spik ldquoRegu-latory effects of lactoferrin and lipopolysaccharide on LFA-1 expression on human peripheral blood mononuclear cellsrdquoArchivum Immunologiae et Therapiae Experimentalis vol 47no 4 pp 257ndash264 1999

Mediators of Inflammation 9

[74] M L Kruzel and M Zimecki ldquoLactoferrin and immunologicdissonance clinical implicationsrdquo Archivum Immunologiae etTherapiae Experimentalis vol 50 no 6 pp 399ndash410 2002

[75] W I Weis M E Taylor and K Drickamer ldquoThe C-type lectinsuperfamily in the immune systemrdquo Immunological Reviewsvol 163 pp 19ndash34 1998

[76] W I Weis K Drickamer and W A Hendrickson ldquoStructureof a C-type mannose-binding protein complexed with an oli-gosacchariderdquo Nature vol 360 no 6400 pp 127ndash134 1992

[77] T Zagulski Z Jarzabek A Zagulska M Jaszczak I EKochanowska and M Zimecki ldquoLactoferrin stimulates killingand clearance of bacteria but does not prevent mortality of dia-betic micerdquo Archivum Immunologiae et Therapiae Experimen-talis vol 49 no 6 pp 431ndash438 2001

[78] M Kruzel Y Harai C Chen and A Castro ldquoRole of lactoferrinin development of systemic inflammatory response syndrome(SIRS)rdquo in Proceedings of the 2nd International ConferenceProgress in Intensive Care Medicine pp 11ndash12 Wroclaw Poland1998

[79] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoThe guta key metabolic organ protected by lactoferrin during experi-mental systemic inflammation in micerdquo in Advances in Lacto-ferrin Research vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Springer New York NY USA 1998

[80] MKruzel T Zagulski andMZimecki ldquoLactoferrin and insult-induced metabolic imbalance in humans and other animalsrdquoin Lactoferrin Structure Function and Applications Proceedingsof the 4th International Conference on Lactoferrin StructureFunction and Applications held in Sapporo Japan 18ndash22 May1999 pp 301ndash311 ICS Elsevier New York NY USA 2000

[81] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoLactofer-rin protects gut mucosal integrity during endotoxemia inducedby lipopolysaccharide in micerdquo Inflammation vol 24 no 1 pp33ndash44 2000

[82] E Zaczynska M Kocięba E Sliwinska and M ZimeckildquoBovine lactoferrin enhances proliferation of human peripheralblood lymphocytes and induces cytokine production in wholeblood culturesrdquo Advances in Clinical and Experimental Medi-cine vol 23 pp 871ndash876 2014

[83] Z Wei T Nishimura and S Yoshida ldquoCharacterization ofglycans in a lactoferrin isoform lactoferrin-ardquo Journal of DairyScience vol 84 no 12 pp 2584ndash2590 2001

[84] B-K Choi J K Actor S Rios et al ldquoRecombinant humanlactoferrin expressed in glycoengineered Pichia pastoris effectof terminal N-acetylneuraminic acid on in vitro secondaryhumoral immune responserdquoGlycoconjugate Journal vol 25 no6 pp 581ndash593 2008

[85] B Samyn-Petit J-PWajdaDubos F Chirat et al ldquoComparativeanalysis of the site-specific N-glycosylation of human lactofer-rin produced in maize and tobacco plantsrdquo European Journal ofBiochemistry vol 270 no 15 pp 3235ndash3242 2003

[86] M Zimecki J Artym M Kocięba M Duk and M L KruzelldquoThe effect of carbohydrate moiety structure on the immu-noregulatory activity of lactoferrin in vitrordquo Cellular ampMolecu-lar Biology Letters vol 19 no 2 pp 284ndash296 2014

[87] J K Actor S-A Hwang and M L Kruzel ldquoLactoferrin as anatural immune modulatorrdquo Current Pharmaceutical Designvol 15 no 17 pp 1956ndash1973 2009

[88] M L Kruzel J K Actor I Boldogh and M ZimeckildquoLactoferrin in health and diseaserdquo Postepy Higieny i MedycynyDoswiadczalnej vol 61 pp 261ndash267 2007

[89] S Baveye E Elass D G Fernig C Blanquart J Mazurier andD Legrand ldquoHuman lactoferrin interacts with soluble CD14and inhibits expression of endothelial adhesion molecules E-selectin and ICAM-1 induced by the CD14-lipopolysaccharidecomplexrdquo Infection and Immunity vol 68 no 12 pp 6519ndash65252000

[90] C S Curran K P Demick and J M Mansfield ldquoLactoferrinactivates macrophages via TLR4-dependent and -independentsignaling pathwaysrdquo Cellular Immunology vol 242 no 1 pp23ndash30 2006

[91] P Puddu D Latorre M Carollo et al ldquoBovine lactoferrincounteracts Toll-Like receptor mediated activation signals inantigen presenting cellsrdquo PLoS ONE vol 6 no 7 Article IDe22504 2011

[92] T M Doherty and M Arditi ldquoTB or not TB That is the ques-tionmdashdoes TLR signaling hold the answerrdquoThe Journal of Clin-ical Investigation vol 114 no 12 pp 1699ndash1703 2004

[93] M A Rahman P Sobia N Gupta L Van Kaer and GDas ldquoMycobacterium tuberculosis subverts the TLR-2mdashMyD88pathway to facilitate its translocation into the cytosolrdquo PLoSONE vol 9 no 1 Article ID e86886 2014

[94] J ArtymM Zimecki andM L Kruzel ldquoEnhanced clearance ofEscherichia coli and Staphylococcus aureus in mice treated withcyclophosphamide and lactoferrinrdquo International Immunophar-macology vol 4 no 9 pp 1149ndash1157 2004

[95] G Chodaczek A Saavedra-Molina A Bacsi M L Kruzel SSur and I Boldogh ldquoIron-mediated dismutation of superoxideanion augments antigen-induced allergic inflammation effectof lactoferrinrdquo Postepy Higieny i Medycyny Doswiadczalnej vol61 pp 268ndash276 2007

[96] M Kruzel Y Harai C Chen and A Castro ldquoThe Gut a keymetabolic organ protected by lactoferrin during experimentalsystemic inflammation in micerdquo in Advances in LactoferrinResearch G Spik D Legrand J Mazurier A Pierce and J-PPerraudin Eds vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Plenum Press 1998

[97] M L Kruzel A Bacsi B Choudhury S Sur and I BoldoghldquoLactoferrin decreases pollen antigen-induced allergic airwayinflammation in a murine model of asthmardquo Immunology vol119 no 2 pp 159ndash166 2006

[98] J K Actor S-A Hwang M Olsen M Zimecki R L HunterJr and M L Kruzel ldquoLactoferrin immunomodulation of DTHresponse in micerdquo International Immunopharmacology vol 2no 4 pp 475ndash486 2002

[99] S-A Hwang M L Kruzel and J K Actor ldquoInfluence of bovinelactoferrin on expression of presentation molecules on BCG-infected bonemarrow derivedmacrophagesrdquo Biochimie vol 91no 1 pp 76ndash85 2009

[100] M Zimecki J Mazurier M Machnicki Z Wieczorek JMontreuil and G Spik ldquoImmunostimulatory activity of lacto-transferrin andmaturation of CD4minus CD8minusmurine thymocytesrdquoImmunology Letters vol 30 no 1 pp 119ndash123 1991

[101] K J Welsh S-A Hwang R L Hunter M L Kruzel and JK Actor ldquoLactoferrin modulation of mycobacterial cord factortrehalose 6-6rsquo-dimycolate induced granulomatous responserdquoTranslational Research vol 156 no 4 pp 207ndash215 2010

[102] M L Bourigault N Segueni S Rose et al ldquoRelative contri-bution of IL-1alpha IL-1beta and TNF to the host responseto Mycobacterium tuberculosis and attenuated M bovis BCGrdquoImmunity Inflammation and Disease vol 1 no 1 pp 47ndash622013

10 Mediators of Inflammation

[103] C Holscher R A Atkinson B Arendse et al ldquoA protective andagonistic function of IL-12p40 in mycobacterial infectionrdquoTheJournal of Immunology vol 167 no 12 pp 6957ndash6966 2001

[104] M Jacobs L Fick N Allie N Brown and B Ryffel ldquoEnhancedimmune response in Mycobacterium bovis bacille calmetteguerin (BCG)-infected IL-10-deficientmicerdquoClinical Chemistryand Laboratory Medicine vol 40 no 9 pp 893ndash902 2002

[105] K J Welsh S-A Hwang S Boyd M L Kruzel R L Hunterand J K Actor ldquoInfluence of oral lactoferrin onMycobacteriumtuberculosis induced immunopathologyrdquo Tuberculosis vol 91supplement 1 pp S105ndashS113 2011

[106] M L Kruzel J K Actor M Zimecki et al ldquoNovel recombinanthuman lactoferrin differential activation of oxidative stressrelated gene expressionrdquo Journal of Biotechnology vol 168 no4 pp 666ndash675 2013

[107] Y Shibata J Gabbard M Yamashita et al ldquoHeat-killedBCG induces biphasic cyclooxygenase 2+ splenic macrophageformationmdashrole of IL-10 and bone marrow precursorsrdquo Journalof Leukocyte Biology vol 80 no 3 pp 590ndash598 2006

[108] H Yamada E Kuroda S Matsumoto T Matsumoto TYamada and U Yamashita ldquoProstaglandin E

2down-regulates

viable Bacille Calmette-Guerin-induced macrophage cytotoxi-city against murine bladder cancer cell MBT-2 in vitrordquoClinicaland Experimental Immunology vol 128 no 1 pp 52ndash58 2002

[109] S Hwang M L Kruzel and J K Actor ldquoEffects of CHO-expressed recombinant lactoferrins on mouse dendritic cellpresentation and functionrdquo Innate Immunity vol 21 no 5 pp553ndash561 2015

[110] K M OShea S-A Hwang and J K Actor ldquoImmune activity ofbcg infected mouse macrophages treated with a novel recom-binant mouse lactoferrinrdquo Annals of Clinical Lab Science vol45 no 5 pp 403ndash410 2015

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Behavioural Neurology

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Disease Markers

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OncologyJournal of

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Oxidative Medicine and Cellular Longevity

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The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

2 Mediators of Inflammation

contributions to factors involved in the development of pro-gressive pathology Furthermore we utilized our knowledgeto design protocols to maximize immune-modulating agentsto investigate the relationship of protection and pathology[21ndash24] An area of study used lactoferrin as an immunemod-ulator to augment the BCG vaccine and relative responsesto mycobacterial antigens [22ndash27] leading to reduction inpathological damage after challenge with virulent organismsThey were the observations made during development ofhuman lactoferrin as a vaccine adjuvant that led us tobelieve it also had utility as an immune modulator to reduceinflammation during granuloma formation [23 28]

Many of the molecular mechanisms involved in develop-ment maintenance and initiation of factors related to myco-bacterial pathology in the lung have been identified A systemof choice is amodel based on the noninfectious proinflamma-tory granuloma induced by the major surface glycolipid frompathogenic organisms developed by multiple investigatorsincluding Retzinger Behling and Hunter [29ndash32] This haspermitted a global understanding of the immunopathologyInjection with cord factor (trehalose 661015840-dimycolate TDM)induces transient pulmonary granulomas that increase innumber and complexity over a 7-day period followed byresolution of pathology [33] Manipulation of host factors hasallowed definition of immune parameters contributing to theinitiation maintenance and resolution phases of response

TheCord Factor Model of theMycobacterial Granuloma Cordfactor (trehalose 661015840-dimycolate TDM) is the most abun-dant glycolipid produced on the surface of mycobacterialorganisms TDM plays multiple roles in the pathogenesis ofMTB [34 35] including the formation of caseation in thelung after infection [36] TDM formulated into an emulsionor placed on beads and injected intravenously into miceinduces a lung pathology that mimics many aspects ofearly MTB infection including granulomatous response andthe production of proinflammatory cytokines The TDMmodel of granuloma formation has been used to elucidatethe immunological factors involved in the granulomatousresponse [37 38] Furthermore TDM can induce activatedforeign body type granulomas in naıve mice [33 39] andhypersensitivity (immune) granulomas in appropriately sen-sitized mice [36 40 41] Therefore this model system isideal for exploring the potential of immunomodulators toalter granuloma structure with perhaps specific utility toextrapolate findings to immune related pathology identifiedduring clinical manifestation of tuberculosis disease

The interactions of mycobacteria and its lipid com-ponents with cell surface antigens to elicit inflammatoryresponses have been characterized [42] and are the subject ofother papers and reviews [43ndash46] Recent discoveries link theC-type lectin mincle [47ndash49] as a prime candidate receptorfor TDM This is augmented in part by interactions withMARCO TLR2 andor CD14 all of which are critical tomediate activity [44] These are combined with internal-ized signaling events that possibly function through Card9-dependentmechanisms [50 51] Suffice to say TDMrsquos interac-tionwith surface ldquoreceptorsrdquo is critical for initiation of cellularactivation of TLR and NOD pathways as well as control of

intracellular trafficking events [52 53] Using knockout miceestablished patterns of cytokine production were found tobe associated with pathology This allowed the delineationof the major molecular events in innate establishment andmaintenance of the TDM induced granuloma [17 33 37 3841 52ndash56] A link was also established to adaptive (T-cellmediated) hypersensitive responses critical for developmentof pathological granulomas [57 58] (Figure 1)

Lactoferrin as a First-Line Defense Protein Lactoferrin is amonomeric 80 kDa single polypeptide chain contained inmost mammalian exocrine secretions including milk tearssaliva and bronchial and intestinal secretions It is alsopresent in the secondary granules of neutrophils It is consid-ered a first-line defense protein involved in protection againstmicrobial infections [59 60] and subsequent development ofsystemic inflammatory response syndrome (SIRS) and sepsis[61ndash67] More recently lactoferrin has been implicated inimmunoregulatory functions [62 68ndash73] and modulation ofvaccine function [22ndash27]

There are two primary forms of human lactoferrinone contained in exocrine secretions and the other presentin secondary granules of neutrophils The two forms areidentical in their amino acid sequence but differ in glycancontent [75 76] While the secreted form is thought tobe involved in the host defense against microbial infec-tion at mucosal sites granulocyticneutrophilic lactoferrinhas notable immunomodulatory function [74] Neutrophiliclactoferrin is an integral part of the cytokine-mediatedcascade during insult-induced metabolic imbalance [77ndash81](Figure 2) with a subset of biological properties uniquefrom lactoferrins produced at other sources by other cellphenotypes [82]

The key to understanding the molecular basis of itsactivities is thought to reside in both patterns of glycosylationand sialylation [83 84] The primary structure of humanLF is characterized by a single polypeptide chain containing692 amino acids organized in two highly homologous lobesdesignated the N- and C-lobe each capable of bindingsingle ferric ion (Fe+++) Lactoferrin is a glycoprotein andin humans the glycans are the N-acetyllactosaminic type12057213-fucosylated on the N-acetyl-glucosamine residue linkedto the peptide chain There are three possible N-linkedglycosylation sites in hLF one at Asn138 a second site atAsn479 and a third site at Asn624 differential utilizationof these sites results in distinct glycosylation variants [85]Many observed activities of LF are dependent upon specificglycosylation patterns For example the immunoregulatoryactivity of LF in humans is dependent on the interaction ofthis glycoprotein with a receptor specific for sialic acid anddirect lymphocyte activation by LF requires sialylation [86]

2 Lactoferrin as an Immune Modulator

21 Lactoferrin A Balance between Potentiation and Media-tion Lactoferrin bridges innate and adaptive immune func-tions by regulating specific target cell responses [87 88] Byacting as a homeostatic modulator lactoferrin can work bothends of the immunological spectrum to increase low response

Mediators of Inflammation 3

TDM

Macrophage

TDM

TDM ComplementIL-6

Initiation phase Maintenance phase

IL-6

Active cortisol Inactive cortisol

Destructive pathology

Hypersensitive regulation

T-cell

Innate Adaptive

11120573HSDI 11120573HSDII

TNF-120572 (C5a)

Figure 1 Mechanisms of TDM-induced granulomas Cytokines and immune-regulatory factors induced by mycobacterial glycolipidTDM play key roles in the development of pathology induced during mycobacterial infection Model systems using isolated TDM allowidentification of host immune function to extrapolate findings of immune related pathology occurring during clinical manifestation oftuberculosis disease [17 33 37 38 41 52ndash58]

Insult RepairAcute inflammation

NeutrophilDendritic cellMacrophage

LFLF

LFLF

Augmentlow antigenic response

Modulateaggressive inflammation

Proinflammatory(TNF120572 IL-6

IL-1120573 and IL-8)

Figure 2 Lactoferrin mediation of insult-induced inflammation Insult defined as infection trauma or chronic illness leads to activation ofmacrophage and dendritic cells Activated neutrophils degranulate at the site of injury and releasemassive amounts of lactoferrin Lactoferrinin turn can either augment low level responses or modulate overaggressive cytokine activity Both effects serve to control inflammation toassist with tissue repair after insult (modified from [74])

or dampen aggressive ones It has direct capacity to regulateboth proinflammatory and anti-inflammatory responses [6263] The utility of such an immune mediator represents anovel therapeutic agent dependent on elicited responses foroutcome [74] A direct example of how lactoferrin canmanip-ulate outcomes of response is exemplified by knowledge thatlactoferrin can bind to the soluble CD14 and the CD14LPScomplex [89] an interaction which has been shown to medi-ate toll-like receptor-4 (TLR-4) pattern recognition events[90] Indeed lactoferrin mediated events related to TLR-4may represent a strong strategy to block excessive antigenpresenting cell activation [91]What remains unknown is howlactoferrin might affect PAMP signaling processes duringtuberculosis infection While some data suggests that Myd88pathways may not be as critical in transition to adaptivedefense against tuberculosis [92] more recent studies suggestthat subversion of theTLR-2-MyD88 pathway is an important

factor in intracellular processing [93] In either case it wouldseem that lactoferrin may function as a regulator in anindependent manner during immune modulation during TBchallenge

We and collaborators investigated lactoferrin to modifyinnate events during inflammation such as those seen dur-ing systemic inflammatory response syndrome and modelsof oxidative stress [78 94ndash97] Our laboratory also spentyears investigating lactoferrin augmentation of the BCGvaccine to protect against subsequent challenge with virulentMTB functioning through strong induction of cell-mediatedimmunity [64 98] and generation of IFN-120574 antigen specificrecall responses [24 26 27 69 98 99] Lactoferrin has aprofound modulatory action on adaptive immune functions[70 73] by promoting maturation of T-cell precursors intocompetent helper cells and differentiation of immature B-cells into efficient antigen presenting cells [100] The innate

4 Mediators of Inflammation

and adaptive events are not mutually exclusive but rathercomplementary and lend credence to utility of lactoferrin forimmune homeostasis in insult-inducedmetabolic disparities

22 Lactoferrin Modulation of the Tuberculoid GranulomaAn examination of lactoferrin to modulate granulomatousresponses was performed in mice injected intravenouslywith TDM A subset of mice were intravenously given 1mgbovine lactoferrin 24 h after TDM challenge Lung tissue wasanalyzed for histological response and for the productionof proinflammatory mediators Treatment with lactoferrinshowed statistically significant fewer and smaller granulomascompared to TDM alone [101] Of importance the protectiveproinflammatory mediators were not statistically diminishedby lactoferrin treatment Nontreated mice demonstrateda granuloma formation that correlated with an increasedproduction of tumor necrosis factor-120572 (TNF-120572) interleukin-(IL-) 1120573 and IL-6This is important as it suggests that innatefunctions critical during infectious disease are maintainedeven though immunopathology is reduced by treatment

In a similar manner the utility of oral delivered lactofer-rin was also examined As previously mentioned mice givenTDM alone also showed marked and statistically significantincreased production of proinflammatory cytokines TNF-120572IL-6 IL-1120573 IL-12p40 and KC (keratinocyte chemoattrac-tant) Oral treatment with recombinant human LF (producedin CHO cells) at 1 or 2mg led to statistically significantreduction (119901 lt 005) in cytokine levels at 7 days after TDMadministration in BALBc and C57BL6 mice (Figure 3 onlyBALBc data shown) This is consistent with reports thatthese proinflammatory cytokines are protective in similarinfectious challenge models [102 103] while molecules suchas IL-10 which are limited after lactoferrin administrationare detrimental during infection [104] In a similar mannerbovine lactoferrin given orally at 1 or 2mg doses also showedstatistical reduction in most cytokines although there wasgreater consistency of response in BALBc mice than thatfor the C57BL6 strain In particular the 1mg dosage ofrecombinant LF showed the most consistency in its ability tocontrol (reduce) production of proinflammatory cytokines

Of major interest the oral delivered human recombinantlactoferrin demonstrated biologically relevant reduction inpathology (Figure 4) similar to that seen in the publishedresults using intravenous administered bovine lactoferrin Infact both the bovine and the recombinant human lactoferrinswere able to reduce inflammation due toTDMchallenge withdoses of 1 or 2mg given on days 4 and 6 after challenge result-ing in clear reduction in granuloma size and frequency Themost effective dose for the recombinant human lactoferrinwas seen when used at 1mg with some efficacy also seengiven as a single dose only on day 4 (data not shown)

Another point of special interest is that the classicalcytokines critical for control of MTB namely TNF-120572 IL-6 and IFN-120574 were not statistically altered by lactoferrintreatment [105] This gave confidence that treatment wouldretain control of organisms after infectious challenge Indeedthis is what was seen in a similar experiment performedusing bovine lactoferrin administered in drinking water(5mgmL) given to mice aerosol infected with virulent

mycobacteria (Erdman strain) [105] While bacterial loadin tissue was slightly reduced the major change was ame-lioration of granulomatous severity It is noteworthy thatbovine lactoferrin reduced bacterial burden accompaniedby an increase in classical proinflammatory responses whiledecreasing overall lung immunopathology [105] Specificallylactoferrin-treated mice increased numbers of CD4+ IFN-120574+ and IL-17 producing cells in the lung It was shown thatlactoferrin by itself was not bactericidal but rather enhancedIFN-120574mediatedMTBkilling bymacrophages in a nitric oxidedependent manner These studies indicate that lactoferrinmay be a novel therapeutic for the treatment of tuberculosisand may be useful in infectious diseases to reduced immune-mediated tissue damage

Finally the molecular mechanism underlying resolutionof the granulomatous lesion is not yet defined However arole for maintaining strong Th1 responses could possibly beattributed to relationships due to COX-2 inhibition by lacto-ferrin in inducedmacrophages (WBCs) [106] and knowledgethat prostaglandin E2 (PGE2) is able to shift T helperresponses during BCG activation of innate macrophages[107] It had been previously demonstrated that the PGE2response could be reversed by a COX-2 inhibitor [108]Therefore lactoferrin functioning as a COX-2 inhibitor couldbe an indirect method to maintain proinflammatory activityof macrophages when infected

3 Conclusions

The studies here indicate that lactoferrin may be a usefuladjunct therapeutic for amelioration of pathological responsein an in vivo model of TB granulomas Of high importancethe change in pathology is accomplished with no loss ofinnate immune function which would be critical for defenseagainst pathogenic organisms A question arises concerningthe utility of using heterologous lactoferrins in the mousemodel Preliminary investigations show that a newly devel-oped recombinant mouse lactoferrin produced in CHO sta-ble cell lines functions in a manner very similar to the bovineand human recombinant to limit immunopathology afteroral delivery Mechanistically it was shown that the mouseand human forms are nearly identical in modulation of den-dritic cell function in response to Bacillus Calmette-Guerin(BCG) [25 109] The dendritic cell population respondswith increased inflammatory cytokines and a shift towardsMHC Class II expression when BCG is combined withthe lactoferrins However more recent investigations revealthat the macrophage population has a unique trend withpotentially decreased antigen presentation and subsequentT-cell stimulatory activity [110] Overall these results givegreat confidence to move forward to test novel recombinantlactoferrins for adjunct clinical therapeutic effects to alterimmunedependent granulomatous pathology occurring dur-ing tuberculosis infection in humansThe studies here woulddrive the next goals to explore novel recombinant lactoferrinsto mediate reductions in lung histopathology and bacterialburden in BSL3 in vivo models of MTB infection Further-more we hypothesize that reduction of TB-induced inflam-matory pathology would also allow enhanced penetration

Mediators of Inflammation 5

2000

1500

1000

500

0

TDM

Treatment

lowast lowast lowast lowast

IL-12

p40

conc

entr

atio

n (p

glu

ng)

Lung IL-12p40

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

6000

2000

3000

4000

5000

1000

0

TDM

Treatment

lowast lowast lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

Lung TNF-120572

IL-6

conc

entr

atio

n (p

glu

ng)

2000

1500

1000

500

0

TDM

Treatment

lowast lowast lowast

Lung IL-6

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

40000

20000

30000

10000

0

TDM

Treatment

lowast lowast lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

IL-1120573

conc

entr

atio

n (p

glu

ng)

Lung IL-1120573

IL-10

conc

entr

atio

n (p

glu

ng)

Lung IL-10300

200

100

0

TDM

Treatment

lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

TNF-120572

conc

entr

atio

n (p

glu

ng)

Figure 3 Proinflammatory cytokines in lungs are reduced in lactoferrin-treated mice after challenge with TDM BALBc mice werechallenged with 25120583g TDM in a water-in-oil emulsion Bovine or recombinant human lactoferrin (1 or 2mg) was orally administered at4 and 6 days after TDM challenge Methods of TDM induction are described elsewhere [38 101] Lungs were isolated on day 7 after TDMchallenge cytokines were assessed by ELISA values are shown per lung for individual mice lowast119901 lt 005 compared to the TDM emulsion alonetreated mice

of antimycobacterial drugs to sites harboring infectiousagents in essence permitting an efficient mechanism totreat TB infection Overall these studies permit confidenceto target clinical implications and test novel strategies to

modulate MTB-induced granulomatous responses to allowmore efficacious delivery of therapeutics to mycobacterialorganisms residing within lung tissue during activemycobac-terial disease states

6 Mediators of Inflammation

BALB

cC5

7BL

6TDM alone TDM + LFNaiumlve

TDM alone TDM + LFNaiumlve

Figure 4 Granuloma response to TDM challenge in oral recombinant human lactoferrin-treated mice Two strains of mice (BALBc andC57BL6) were challenged with 25120583g TDM in a water-in-oil emulsion as described [38 101] Human lactoferrin (recombinant 1mg) wasgiven orally at 4 and 6 days after TDM challenge Comparisons are made to naıve mice and to mice only administered TDM HampE staining100x Representative sections from mice at 7 days after TDM challenge119873 ge 8 per experimental group

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

This work was supported in part by NIH Grants 1R41-AI117990-01 and R42-AI051050-05The authors acknowledgePharmaReview Corp (Houston TX) for their kind gifts oflactoferrins and thank Marian L Kruzel PhD and Shen-An Hwang PhD for their insights into the research projectThe authors also thank Ron Gadot and Ai Yamazaki for theirtechnical contributions to the research presented

References

[1] P E M Fine ldquoHerd immunity history theory practicerdquo Epide-miologic Reviews vol 15 no 2 pp 265ndash302 1993

[2] L Ramakrishnan ldquoRevisiting the role of the granuloma intuberculosisrdquo Nature Reviews Immunology vol 12 no 5 pp352ndash366 2012

[3] M Silva Miranda A Breiman S Allain F Deknuydt andF Altare ldquoThe tuberculous granuloma an unsuccessful hostdefence mechanism providing a safety shelter for the bacteriardquoClinical amp Developmental Immunology vol 2012 Article ID139127 14 pages 2012

[4] S A Khader J Rangel-Moreno J J Fountain et al ldquoIn amurinetuberculosis model the absence of homeostatic chemokinesdelays granuloma formation and protective immunityrdquo Journalof Immunology vol 183 no 12 pp 8004ndash8014 2009

[5] AMCooper and S A Khader ldquoThe role of cytokines in the ini-tiation expansion and control of cellular immunity to tubercu-losisrdquo Immunological Reviews vol 226 no 1 pp 191ndash204 2008

[6] IMOrme and R J Basaraba ldquoThe formation of the granulomain tuberculosis infectionrdquo Seminars in Immunology vol 26 no6 pp 601ndash609 2014

[7] I M Orme R T Robinson and A M Cooper ldquoThe balancebetween protective and pathogenic immune responses in theTB-infected lungrdquo Nature Immunology vol 16 no 1 pp 57ndash632014

[8] J K Actor M Olsen C Jagannath and R L Hunter ldquoRela-tionship of survival organism containment and granulomaformation in acute murine tuberculosisrdquo Journal of Interferonand Cytokine Research vol 19 no 10 pp 1183ndash1193 1999

[9] J L Flynn and J Chan ldquoWhatrsquos good for the host is good for thebugrdquo Trends in Microbiology vol 13 no 3 pp 98ndash102 2005

[10] D G Russell ldquoMycobacterium tuberculosis and the intimatediscourse of a chronic infectionrdquo Immunological Reviews vol240 no 1 pp 252ndash268 2011

[11] R L Hunter ldquoPathology of post primary tuberculosis of thelung an illustrated critical reviewrdquo Tuberculosis vol 91 no 6pp 497ndash509 2011

[12] R L Hunter J K Actor S-A Hwang V Karev and C Jagan-nath ldquoPathogenesis of post primary tuberculosis immunity andhypersensitivity in the development of cavitiesrdquoAnnals of Clini-cal and Laboratory Science vol 44 no 4 pp 365ndash387 2014

[13] K J Welsh R L Hunter and J K Actor ldquoTrehalose 661015840-dimy-colatemdasha coat to regulate tuberculosis immunopathogenesisrdquoTuberculosis vol 93 supplement pp S3ndashS9 2013

[14] M-J Kim H C Wainwright M Locketz et al ldquoCaseation ofhuman tuberculosis granulomas correlates with elevated hostlipid metabolismrdquo EMBO Molecular Medicine vol 2 no 7 pp258ndash274 2010

[15] J K Actor E Breij R A Wetsel H Hoffmann R L HunterJr and C Jagannath ldquoA role for complement C5 in organ-ism containment and granulomatous response during murine

Mediators of Inflammation 7

tuberculosisrdquo Scandinavian Journal of Immunology vol 53 no5 pp 464ndash474 2001

[16] J K Actor C D Leonard V E Watson et al ldquoCytokinemRNAexpression and serumcortisol evaluation duringmurinelung inflammation induced by Mycobacterium tuberculosisrdquoCombinatorial Chemistry and High Throughput Screening vol3 no 4 pp 343ndash351 2000

[17] C W Borders A Courtney K Ronen et al ldquoRequisite role forcomplement C5 and the C5a receptor in granulomatous res-ponse to mycobacterial glycolipid trehalose 661015840-dimycolaterdquoScandinavian Journal of Immunology vol 62 no 2 pp 123ndash1302005

[18] R L Hunter C Jagannath and J K Actor ldquoPathology of post-primary tuberculosis in humans and mice contradiction oflong-held beliefsrdquo Tuberculosis vol 87 no 4 pp 267ndash278 2007

[19] C Jagannath H Hoffmann E Sepulveda J K Actor R AWetsel and R L Hunter Jr ldquoHypersusceptibility of AJ miceto tuberculosis is in part due to a deficiency of the fifth comple-ment component (C5)rdquo Scandinavian Journal of Immunologyvol 52 no 4 pp 369ndash379 2000

[20] V E Watson L L Hill L B Owen-Schaub et al ldquoApoptosis inMycobacterium tuberculosis infection in mice exhibiting variedimmunopathologyrdquo Journal of Pathology vol 190 no 2 pp 211ndash220 2000

[21] R H Copenhaver E Sepulveda L Y Armitige et al ldquoAmutantof Mycobacterium tuberculosis H37Rv that lacks expressionof antigen 85A is attenuated in mice but retains vaccinogenicpotentialrdquo Infection and Immunity vol 72 no 12 pp 7084ndash7095 2004

[22] S-A Hwang R Arora M L Kruzel and J K Actor ldquoLactofer-rin enhances efficacy of the BCG vaccine comparison betweentwo inbred mice strains (C57BL6 and BALBc)rdquo Tuberculosisvol 89 supplement 1 pp S49ndashS54 2009

[23] S-A Hwang K Wilk M L Kruzel and J K Actor ldquoA novelrecombinant human lactoferrin augments the BCG vaccine andprotects alveolar integrity upon infection with Mycobacteriumtuberculosis in micerdquo Vaccine vol 27 no 23 pp 3026ndash30342009

[24] S-A Hwang K M Wilk M Budnicka et al ldquoLactoferrinenhanced efficacy of the BCG vaccine to generate host protec-tive responses against challenge with virulent Mycobacteriumtuberculosisrdquo Vaccine vol 25 no 37-38 pp 6730ndash6743 2007

[25] S-A Hwang and J K Actor ldquoLactoferrin modulation of BCG-infected dendritic cell functionsrdquo International Immunologyvol 21 no 10 pp 1185ndash1197 2009

[26] S-A Hwang M L Kruzel and J K Actor ldquoLactoferrin aug-ments BCG vaccine efficacy to generate T helper response andsubsequent protection against challenge with virulentMycobac-terium tuberculosisrdquo International Immunopharmacology vol 5no 3 pp 591ndash599 2005

[27] K M Wilk S-A Hwang and J K Actor ldquoLactoferrin mod-ulation of antigen-presenting-cell response to BCG infectionrdquoPostepy Higieny i Medycyny Doswiadczalnej vol 61 pp 277ndash282 2007

[28] S A Hwang M L Kruzel and J K Actor ldquoCHO expressedrecombinant human lactoferrin as an adjuvant for BCGrdquoInternational Journal of Immunopathology and Pharmacologyvol 28 no 4 pp 452ndash468 2015

[29] R L Perez J Roman G W Staton Jr and R L HunterldquoExtravascular coagulation and fibrinolysis in murine lunginflammation induced by the mycobacterial cord factor

trehalose-66rsquo-dimycolaterdquoAmerican Journal of Respiratory andCritical Care Medicine vol 149 no 2 pp 510ndash518 1994

[30] C A Behling R L Perez M R Kidd G W Staton Jr and RL Hunter ldquoInduction of pulmonary granulomas macrophageprocoagulant activity and tumor necrosis factor-alpha by tre-halose glycolipidsrdquo Annals of Clinical and Laboratory Sciencevol 23 no 4 pp 256ndash266 1993

[31] G S Retzinger S C Meredith R L Hunter K Takayama andF J Kezdy ldquoIdentification of the physiologically active state ofthe mycobacterial glycolipid trehalose 661015840-dimycolate and therole of fibrinogen in the biologic activities of trehalose 661015840-dimycolate monolayersrdquo Journal of Immunology vol 129 no 2pp 735ndash744 1982

[32] G S Retzinger S C Meredith K Takayama R L Hunter andF J Kezdy ldquoThe role of surface in the biological activities oftrehalose 661015840-dimycolate Surface properties and developmentof amodel systemrdquoThe Journal of Biological Chemistry vol 256no 15 pp 8208ndash8216 1981

[33] R L Perez J Roman S Roser et al ldquoCytokinemessage and pro-tein expression during lung granuloma formation and resolu-tion induced by the mycobacterial cord factor trehalose-661015840-dimycolaterdquo Journal of Interferon amp Cytokine Research vol 20no 9 pp 795ndash804 2000

[34] R L Hunter M R Olsen C Jagannath and J K Actor ldquoMult-iple roles of cord factor in the pathogenesis of primary sec-ondary and cavitary tuberculosis including a revised descrip-tion of the pathology of secondary diseaserdquo Annals of Clinicaland Laboratory Science vol 36 no 4 pp 371ndash386 2006

[35] J K Actor ldquoMycobacterial TDM a coat to modulate post pri-mary pathogenesisrdquoMycobacterial Diseases vol 2 no 1 articlee106 2012

[36] R L HunterMOlsen C Jagannath and J K Actor ldquoTrehalose661015840-dimycolate and lipid in the pathogenesis of caseatinggranulomas of tuberculosis in micerdquo The American Journal ofPathology vol 168 no 4 pp 1249ndash1261 2006

[37] A N Abbott T V Guidry K J Welsh et al ldquo11Beta-hydroxy-steroid dehydrogenases are regulated during the pulmonarygranulomatous response to the mycobacterial glycolipidtrehalose-661015840-dimycolaterdquo NeuroImmunoModulation vol 16no 3 pp 147ndash154 2009

[38] K J Welsh A N Abbott S-A Hwang et al ldquoA role for tumournecrosis factor-120572 complement C5 and interleukin-6 in theinitiation and development of the mycobacterial cord factortrehalose 661015840-dimycolate induced granulomatous responserdquoMicrobiology vol 154 no 6 pp 1813ndash1824 2008

[39] A Bekierkunst and E Yarkoni ldquoGranulomatous hypersensitiv-ity to trehalose-66rsquo-dimycolate (cord factor) in mice infectedwith BCGrdquo Infection and Immunity vol 7 no 4 pp 631ndash6381973

[40] H Yamagami TMatsumoto N Fujiwara et al ldquoTrehalose 661015840-dimycolate (cord factor) ofMycobacterium tuberculosis inducesforeign-body- and hypersensitivity-type granulomas in micerdquoInfection and Immunity vol 69 no 2 pp 810ndash815 2001

[41] T VGuidry R LHunter Jr and J K Actor ldquoMycobacterial gly-colipid trehalose 661015840-dimycolate-induced hypersensitive gran-ulomas contribution of CD4+ lymphocytesrdquoMicrobiology vol153 no 10 pp 3360ndash3369 2007

[42] E Rhoades F-F Hsu J B Torrelles J Turk D Chatterjee andD G Russell ldquoIdentification and macrophage-activating activ-ity of glycolipids released from intracellular Mycobacteriumbovis BCGrdquoMolecular Microbiology vol 48 no 4 pp 875ndash8882003

8 Mediators of Inflammation

[43] MMHossain andM-N Norazmi ldquoPattern recognition recep-tors and cytokines in Mycobacterium tuberculosis infectionmdashthe double-edged swordrdquo BioMed Research International vol2013 Article ID 179174 18 pages 2013

[44] D M E Bowdish K Sakamoto M-J Kim et al ldquoMARCOTLR2 and CD14 are required for macrophage cytokine res-ponses to mycobacterial trehalose dimycolate and Mycobac-terium tuberculosisrdquo PLoS Pathogens vol 5 no 6 Article IDe1000474 2009

[45] E Mortaz I M Adcock P Tabarsi et al ldquoInteraction of patternrecognition receptors withMycobacterium tuberculosisrdquo Journalof Clinical Immunology vol 35 no 1 pp 1ndash10 2015

[46] D G Russell ldquoWho puts the tubercle in tuberculosisrdquo NatureReviews Microbiology vol 5 no 1 pp 39ndash47 2007

[47] H Schoenen B Bodendorfer K Hitchens et al ldquoCutting Edgemincle is essential for recognition and adjuvanticity of themycobacterial cord factor and its synthetic analog trehalose-dibehenaterdquo Journal of Immunology vol 184 no 6 pp 2756ndash2760 2010

[48] E Ishikawa T Ishikawa Y S Morita et al ldquoDirect recognitionof themycobacterial glycolipid trehalose dimycolate by C-typelectinMinclerdquo Journal of ExperimentalMedicine vol 206 no 13pp 2879ndash2888 2009

[49] I Matsunaga and D B Moody ldquoMincle is a long soughtreceptor for mycobacterial cord factorrdquo Journal of ExperimentalMedicine vol 206 no 13 pp 2865ndash2868 2009

[50] K Werninghaus A Babiak O Gross et al ldquoAdjuvanticity ofa synthetic cord factor analogue for subunit Mycobacteriumtuberculosis vaccination requires FclR120574-Syk-Card9-dependentinnate immune activationrdquo Journal of Experimental Medicinevol 206 no 1 pp 89ndash97 2009

[51] S LeibundGut-LandmannOGrossM J Robinson et al ldquoSyk-and CARD9-dependent coupling of innate immunity to theinduction of T helper cells that produce interleukin 17rdquo NatureImmunology vol 8 no 6 pp 630ndash638 2007

[52] J Indrigo R L Hunter Jr and J K Actor ldquoInfluence of tre-halose 661015840-dimycolate (TDM) during mycobacterial infectionof bone marrow macrophagesrdquoMicrobiology vol 148 no 7 pp1991ndash1998 2002

[53] J Indrigo R L Hunter Jr and J K Actor ldquoCord factortrehalose 661015840-dimycolate (TDM) mediates trafficking eventsduringmycobacterial infection ofmurinemacrophagesrdquoMicro-biology vol 149 no 8 pp 2049ndash2059 2003

[54] J K Actor M Olsen R L Hunter Jr and Y-J Geng ldquoDys-regulated response to mycobacterial cord factor trehalose-661015840-dimycolate inCD1Dminusminusmicerdquo Journal of Interferon andCytokineResearch vol 21 no 12 pp 1089ndash1096 2001

[55] J K Actor J Indrigo C M Beachdel et al ldquoMycobacterial gly-colipid cord factor trehalose 661015840-dimycolate causes a decreasein serum cortisol during the granulomatous responserdquo Neuro-ImmunoModulation vol 10 no 5 pp 270ndash282 2002

[56] A N Abbott K J Welsh S-A Hwang et al ldquoIL-6 mediates11120573hsd type 2 to effect progression of the mycobacterial cordfactor trehalose 661015840-dimycolate-induced granulomatous res-ponserdquo NeuroImmunoModulation vol 18 no 4 pp 212ndash2252011

[57] T V Guidry M Olsen K-S Kil R L Hunter Jr Y-J Gengand J K Actor ldquoFailure of CD1Dminusminusmice to elicit hypersensitivegranulomas to mycobacterial cord factor trehalose 661015840-dimy-colaterdquo Journal of Interferon amp Cytokine Research vol 24 no 6pp 362ndash371 2004

[58] TVGuidry R LHunter Jr and J KActor ldquoCD3+ cells transferthe hypersensitive granulomatous response to mycobacterialglycolipid trehalose 661015840-dimycolate in micerdquoMicrobiology vol152 no 12 pp 3765ndash3775 2006

[59] B Lonnerdal and S Iyer ldquoLactoferrin molecular structure andbiological functionrdquoAnnual Review of Nutrition vol 15 pp 93ndash110 1995

[60] L Sanchez M Calvo and J H Brock ldquoBiological role of lacto-ferrinrdquo Archives of Disease in Childhood vol 67 no 5 pp 657ndash661 1992

[61] P Manzoni M Rinaldi S Cattani et al ldquoBovine lactoferrinsupplementation for prevention of late-onset sepsis in verylow-birth-weight neonates a randomized trialrdquo Journal of theAmerican Medical Association vol 302 no 13 pp 1421ndash14282009

[62] S Baveye E Elass J Mazurier G Spik and D Legrand ldquoLacto-ferrin a multifunctional glycoprotein involved in the modula-tion of the inflammatory processrdquoClinical Chemistry and Labo-ratory Medicine vol 37 no 3 pp 281ndash286 1999

[63] R D Baynes and W R Bezwoda ldquoLactoferrin and the inflam-matory responserdquo Advances in Experimental Medicine andBiology vol 357 pp 133ndash141 1994

[64] M Kocieba M Zimecki M Kruzel and J Actor ldquoThe adjuvantactivity of lactoferrin in the generation of DTH to ovalbumincan be inhibited by bovine serum albumin bearing alpha-D-mannopyranosyl residuesrdquoCellularampMolecular Biology Lettersvol 7 no 4 pp 1131ndash1136 2002

[65] C G Turin A Zea-Vera A Pezo et al ldquoLactoferrin for pre-vention of neonatal sepsisrdquo Biometals vol 27 no 5 pp 1007ndash1016 2014

[66] T Siqueiros-Cendon S Arevalo-Gallegos B F Iglesias-Figueroa I A Garcıa-Montoya J Salazar-Martınez and QRascon-Cruz ldquoImmunomodulatory effects of lactoferrinrdquo ActaPharmacologica Sinica vol 35 no 5 pp 557ndash566 2014

[67] D Latorre F Berlutti P Valenti S Gessani and P Puddu ldquoLFimmunomodulatory strategies mastering bacterial endotoxinrdquoBiochemistry and Cell Biology vol 90 no 3 pp 269ndash278 2012

[68] D Legrand ldquoLactoferrin a key molecule in immune andinflammatory processesrdquo Biochemistry and Cell Biology vol 90no 3 pp 252ndash268 2012

[69] S-A Hwang K M Wilk Y A Bangale M L Kruzel and JK Actor ldquoLactoferrin modulation of IL-12 and IL-10 responsefrom activated murine leukocytesrdquo Medical Microbiology andImmunology vol 196 no 3 pp 171ndash180 2007

[70] M Zimecki J Mazurier G Spik and J A Kapp ldquoHuman lacto-ferrin induces phenotypic and functional changes in murinesplenic B cellsrdquo Immunology vol 86 no 1 pp 122ndash127 1995

[71] M Zimecki J Artym G Chodaczek et al ldquoImmunoregulatoryfunction of lactoferrin in immunosuppressed and autoimmuneanimalsrdquo Postepy Higieny i Medycyny Doswiadczalnej vol 61pp 283ndash287 2007

[72] M Zimecki and M L Kruzel ldquoMilk-derived proteins andpeptides of potential therapeutic and nutritive valuerdquo Journal ofExperimental Therapeutics and Oncology vol 6 no 2 pp 89ndash106 2007

[73] M Zimecki RMiędzybrodzki J Mazurier andG Spik ldquoRegu-latory effects of lactoferrin and lipopolysaccharide on LFA-1 expression on human peripheral blood mononuclear cellsrdquoArchivum Immunologiae et Therapiae Experimentalis vol 47no 4 pp 257ndash264 1999

Mediators of Inflammation 9

[74] M L Kruzel and M Zimecki ldquoLactoferrin and immunologicdissonance clinical implicationsrdquo Archivum Immunologiae etTherapiae Experimentalis vol 50 no 6 pp 399ndash410 2002

[75] W I Weis M E Taylor and K Drickamer ldquoThe C-type lectinsuperfamily in the immune systemrdquo Immunological Reviewsvol 163 pp 19ndash34 1998

[76] W I Weis K Drickamer and W A Hendrickson ldquoStructureof a C-type mannose-binding protein complexed with an oli-gosacchariderdquo Nature vol 360 no 6400 pp 127ndash134 1992

[77] T Zagulski Z Jarzabek A Zagulska M Jaszczak I EKochanowska and M Zimecki ldquoLactoferrin stimulates killingand clearance of bacteria but does not prevent mortality of dia-betic micerdquo Archivum Immunologiae et Therapiae Experimen-talis vol 49 no 6 pp 431ndash438 2001

[78] M Kruzel Y Harai C Chen and A Castro ldquoRole of lactoferrinin development of systemic inflammatory response syndrome(SIRS)rdquo in Proceedings of the 2nd International ConferenceProgress in Intensive Care Medicine pp 11ndash12 Wroclaw Poland1998

[79] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoThe guta key metabolic organ protected by lactoferrin during experi-mental systemic inflammation in micerdquo in Advances in Lacto-ferrin Research vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Springer New York NY USA 1998

[80] MKruzel T Zagulski andMZimecki ldquoLactoferrin and insult-induced metabolic imbalance in humans and other animalsrdquoin Lactoferrin Structure Function and Applications Proceedingsof the 4th International Conference on Lactoferrin StructureFunction and Applications held in Sapporo Japan 18ndash22 May1999 pp 301ndash311 ICS Elsevier New York NY USA 2000

[81] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoLactofer-rin protects gut mucosal integrity during endotoxemia inducedby lipopolysaccharide in micerdquo Inflammation vol 24 no 1 pp33ndash44 2000

[82] E Zaczynska M Kocięba E Sliwinska and M ZimeckildquoBovine lactoferrin enhances proliferation of human peripheralblood lymphocytes and induces cytokine production in wholeblood culturesrdquo Advances in Clinical and Experimental Medi-cine vol 23 pp 871ndash876 2014

[83] Z Wei T Nishimura and S Yoshida ldquoCharacterization ofglycans in a lactoferrin isoform lactoferrin-ardquo Journal of DairyScience vol 84 no 12 pp 2584ndash2590 2001

[84] B-K Choi J K Actor S Rios et al ldquoRecombinant humanlactoferrin expressed in glycoengineered Pichia pastoris effectof terminal N-acetylneuraminic acid on in vitro secondaryhumoral immune responserdquoGlycoconjugate Journal vol 25 no6 pp 581ndash593 2008

[85] B Samyn-Petit J-PWajdaDubos F Chirat et al ldquoComparativeanalysis of the site-specific N-glycosylation of human lactofer-rin produced in maize and tobacco plantsrdquo European Journal ofBiochemistry vol 270 no 15 pp 3235ndash3242 2003

[86] M Zimecki J Artym M Kocięba M Duk and M L KruzelldquoThe effect of carbohydrate moiety structure on the immu-noregulatory activity of lactoferrin in vitrordquo Cellular ampMolecu-lar Biology Letters vol 19 no 2 pp 284ndash296 2014

[87] J K Actor S-A Hwang and M L Kruzel ldquoLactoferrin as anatural immune modulatorrdquo Current Pharmaceutical Designvol 15 no 17 pp 1956ndash1973 2009

[88] M L Kruzel J K Actor I Boldogh and M ZimeckildquoLactoferrin in health and diseaserdquo Postepy Higieny i MedycynyDoswiadczalnej vol 61 pp 261ndash267 2007

[89] S Baveye E Elass D G Fernig C Blanquart J Mazurier andD Legrand ldquoHuman lactoferrin interacts with soluble CD14and inhibits expression of endothelial adhesion molecules E-selectin and ICAM-1 induced by the CD14-lipopolysaccharidecomplexrdquo Infection and Immunity vol 68 no 12 pp 6519ndash65252000

[90] C S Curran K P Demick and J M Mansfield ldquoLactoferrinactivates macrophages via TLR4-dependent and -independentsignaling pathwaysrdquo Cellular Immunology vol 242 no 1 pp23ndash30 2006

[91] P Puddu D Latorre M Carollo et al ldquoBovine lactoferrincounteracts Toll-Like receptor mediated activation signals inantigen presenting cellsrdquo PLoS ONE vol 6 no 7 Article IDe22504 2011

[92] T M Doherty and M Arditi ldquoTB or not TB That is the ques-tionmdashdoes TLR signaling hold the answerrdquoThe Journal of Clin-ical Investigation vol 114 no 12 pp 1699ndash1703 2004

[93] M A Rahman P Sobia N Gupta L Van Kaer and GDas ldquoMycobacterium tuberculosis subverts the TLR-2mdashMyD88pathway to facilitate its translocation into the cytosolrdquo PLoSONE vol 9 no 1 Article ID e86886 2014

[94] J ArtymM Zimecki andM L Kruzel ldquoEnhanced clearance ofEscherichia coli and Staphylococcus aureus in mice treated withcyclophosphamide and lactoferrinrdquo International Immunophar-macology vol 4 no 9 pp 1149ndash1157 2004

[95] G Chodaczek A Saavedra-Molina A Bacsi M L Kruzel SSur and I Boldogh ldquoIron-mediated dismutation of superoxideanion augments antigen-induced allergic inflammation effectof lactoferrinrdquo Postepy Higieny i Medycyny Doswiadczalnej vol61 pp 268ndash276 2007

[96] M Kruzel Y Harai C Chen and A Castro ldquoThe Gut a keymetabolic organ protected by lactoferrin during experimentalsystemic inflammation in micerdquo in Advances in LactoferrinResearch G Spik D Legrand J Mazurier A Pierce and J-PPerraudin Eds vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Plenum Press 1998

[97] M L Kruzel A Bacsi B Choudhury S Sur and I BoldoghldquoLactoferrin decreases pollen antigen-induced allergic airwayinflammation in a murine model of asthmardquo Immunology vol119 no 2 pp 159ndash166 2006

[98] J K Actor S-A Hwang M Olsen M Zimecki R L HunterJr and M L Kruzel ldquoLactoferrin immunomodulation of DTHresponse in micerdquo International Immunopharmacology vol 2no 4 pp 475ndash486 2002

[99] S-A Hwang M L Kruzel and J K Actor ldquoInfluence of bovinelactoferrin on expression of presentation molecules on BCG-infected bonemarrow derivedmacrophagesrdquo Biochimie vol 91no 1 pp 76ndash85 2009

[100] M Zimecki J Mazurier M Machnicki Z Wieczorek JMontreuil and G Spik ldquoImmunostimulatory activity of lacto-transferrin andmaturation of CD4minus CD8minusmurine thymocytesrdquoImmunology Letters vol 30 no 1 pp 119ndash123 1991

[101] K J Welsh S-A Hwang R L Hunter M L Kruzel and JK Actor ldquoLactoferrin modulation of mycobacterial cord factortrehalose 6-6rsquo-dimycolate induced granulomatous responserdquoTranslational Research vol 156 no 4 pp 207ndash215 2010

[102] M L Bourigault N Segueni S Rose et al ldquoRelative contri-bution of IL-1alpha IL-1beta and TNF to the host responseto Mycobacterium tuberculosis and attenuated M bovis BCGrdquoImmunity Inflammation and Disease vol 1 no 1 pp 47ndash622013

10 Mediators of Inflammation

[103] C Holscher R A Atkinson B Arendse et al ldquoA protective andagonistic function of IL-12p40 in mycobacterial infectionrdquoTheJournal of Immunology vol 167 no 12 pp 6957ndash6966 2001

[104] M Jacobs L Fick N Allie N Brown and B Ryffel ldquoEnhancedimmune response in Mycobacterium bovis bacille calmetteguerin (BCG)-infected IL-10-deficientmicerdquoClinical Chemistryand Laboratory Medicine vol 40 no 9 pp 893ndash902 2002

[105] K J Welsh S-A Hwang S Boyd M L Kruzel R L Hunterand J K Actor ldquoInfluence of oral lactoferrin onMycobacteriumtuberculosis induced immunopathologyrdquo Tuberculosis vol 91supplement 1 pp S105ndashS113 2011

[106] M L Kruzel J K Actor M Zimecki et al ldquoNovel recombinanthuman lactoferrin differential activation of oxidative stressrelated gene expressionrdquo Journal of Biotechnology vol 168 no4 pp 666ndash675 2013

[107] Y Shibata J Gabbard M Yamashita et al ldquoHeat-killedBCG induces biphasic cyclooxygenase 2+ splenic macrophageformationmdashrole of IL-10 and bone marrow precursorsrdquo Journalof Leukocyte Biology vol 80 no 3 pp 590ndash598 2006

[108] H Yamada E Kuroda S Matsumoto T Matsumoto TYamada and U Yamashita ldquoProstaglandin E

2down-regulates

viable Bacille Calmette-Guerin-induced macrophage cytotoxi-city against murine bladder cancer cell MBT-2 in vitrordquoClinicaland Experimental Immunology vol 128 no 1 pp 52ndash58 2002

[109] S Hwang M L Kruzel and J K Actor ldquoEffects of CHO-expressed recombinant lactoferrins on mouse dendritic cellpresentation and functionrdquo Innate Immunity vol 21 no 5 pp553ndash561 2015

[110] K M OShea S-A Hwang and J K Actor ldquoImmune activity ofbcg infected mouse macrophages treated with a novel recom-binant mouse lactoferrinrdquo Annals of Clinical Lab Science vol45 no 5 pp 403ndash410 2015

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Behavioural Neurology

EndocrinologyInternational Journal of

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Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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OncologyJournal of

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Oxidative Medicine and Cellular Longevity

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The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Research and TreatmentAIDS

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Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Mediators of Inflammation 3

TDM

Macrophage

TDM

TDM ComplementIL-6

Initiation phase Maintenance phase

IL-6

Active cortisol Inactive cortisol

Destructive pathology

Hypersensitive regulation

T-cell

Innate Adaptive

11120573HSDI 11120573HSDII

TNF-120572 (C5a)

Figure 1 Mechanisms of TDM-induced granulomas Cytokines and immune-regulatory factors induced by mycobacterial glycolipidTDM play key roles in the development of pathology induced during mycobacterial infection Model systems using isolated TDM allowidentification of host immune function to extrapolate findings of immune related pathology occurring during clinical manifestation oftuberculosis disease [17 33 37 38 41 52ndash58]

Insult RepairAcute inflammation

NeutrophilDendritic cellMacrophage

LFLF

LFLF

Augmentlow antigenic response

Modulateaggressive inflammation

Proinflammatory(TNF120572 IL-6

IL-1120573 and IL-8)

Figure 2 Lactoferrin mediation of insult-induced inflammation Insult defined as infection trauma or chronic illness leads to activation ofmacrophage and dendritic cells Activated neutrophils degranulate at the site of injury and releasemassive amounts of lactoferrin Lactoferrinin turn can either augment low level responses or modulate overaggressive cytokine activity Both effects serve to control inflammation toassist with tissue repair after insult (modified from [74])

or dampen aggressive ones It has direct capacity to regulateboth proinflammatory and anti-inflammatory responses [6263] The utility of such an immune mediator represents anovel therapeutic agent dependent on elicited responses foroutcome [74] A direct example of how lactoferrin canmanip-ulate outcomes of response is exemplified by knowledge thatlactoferrin can bind to the soluble CD14 and the CD14LPScomplex [89] an interaction which has been shown to medi-ate toll-like receptor-4 (TLR-4) pattern recognition events[90] Indeed lactoferrin mediated events related to TLR-4may represent a strong strategy to block excessive antigenpresenting cell activation [91]What remains unknown is howlactoferrin might affect PAMP signaling processes duringtuberculosis infection While some data suggests that Myd88pathways may not be as critical in transition to adaptivedefense against tuberculosis [92] more recent studies suggestthat subversion of theTLR-2-MyD88 pathway is an important

factor in intracellular processing [93] In either case it wouldseem that lactoferrin may function as a regulator in anindependent manner during immune modulation during TBchallenge

We and collaborators investigated lactoferrin to modifyinnate events during inflammation such as those seen dur-ing systemic inflammatory response syndrome and modelsof oxidative stress [78 94ndash97] Our laboratory also spentyears investigating lactoferrin augmentation of the BCGvaccine to protect against subsequent challenge with virulentMTB functioning through strong induction of cell-mediatedimmunity [64 98] and generation of IFN-120574 antigen specificrecall responses [24 26 27 69 98 99] Lactoferrin has aprofound modulatory action on adaptive immune functions[70 73] by promoting maturation of T-cell precursors intocompetent helper cells and differentiation of immature B-cells into efficient antigen presenting cells [100] The innate

4 Mediators of Inflammation

and adaptive events are not mutually exclusive but rathercomplementary and lend credence to utility of lactoferrin forimmune homeostasis in insult-inducedmetabolic disparities

22 Lactoferrin Modulation of the Tuberculoid GranulomaAn examination of lactoferrin to modulate granulomatousresponses was performed in mice injected intravenouslywith TDM A subset of mice were intravenously given 1mgbovine lactoferrin 24 h after TDM challenge Lung tissue wasanalyzed for histological response and for the productionof proinflammatory mediators Treatment with lactoferrinshowed statistically significant fewer and smaller granulomascompared to TDM alone [101] Of importance the protectiveproinflammatory mediators were not statistically diminishedby lactoferrin treatment Nontreated mice demonstrateda granuloma formation that correlated with an increasedproduction of tumor necrosis factor-120572 (TNF-120572) interleukin-(IL-) 1120573 and IL-6This is important as it suggests that innatefunctions critical during infectious disease are maintainedeven though immunopathology is reduced by treatment

In a similar manner the utility of oral delivered lactofer-rin was also examined As previously mentioned mice givenTDM alone also showed marked and statistically significantincreased production of proinflammatory cytokines TNF-120572IL-6 IL-1120573 IL-12p40 and KC (keratinocyte chemoattrac-tant) Oral treatment with recombinant human LF (producedin CHO cells) at 1 or 2mg led to statistically significantreduction (119901 lt 005) in cytokine levels at 7 days after TDMadministration in BALBc and C57BL6 mice (Figure 3 onlyBALBc data shown) This is consistent with reports thatthese proinflammatory cytokines are protective in similarinfectious challenge models [102 103] while molecules suchas IL-10 which are limited after lactoferrin administrationare detrimental during infection [104] In a similar mannerbovine lactoferrin given orally at 1 or 2mg doses also showedstatistical reduction in most cytokines although there wasgreater consistency of response in BALBc mice than thatfor the C57BL6 strain In particular the 1mg dosage ofrecombinant LF showed the most consistency in its ability tocontrol (reduce) production of proinflammatory cytokines

Of major interest the oral delivered human recombinantlactoferrin demonstrated biologically relevant reduction inpathology (Figure 4) similar to that seen in the publishedresults using intravenous administered bovine lactoferrin Infact both the bovine and the recombinant human lactoferrinswere able to reduce inflammation due toTDMchallenge withdoses of 1 or 2mg given on days 4 and 6 after challenge result-ing in clear reduction in granuloma size and frequency Themost effective dose for the recombinant human lactoferrinwas seen when used at 1mg with some efficacy also seengiven as a single dose only on day 4 (data not shown)

Another point of special interest is that the classicalcytokines critical for control of MTB namely TNF-120572 IL-6 and IFN-120574 were not statistically altered by lactoferrintreatment [105] This gave confidence that treatment wouldretain control of organisms after infectious challenge Indeedthis is what was seen in a similar experiment performedusing bovine lactoferrin administered in drinking water(5mgmL) given to mice aerosol infected with virulent

mycobacteria (Erdman strain) [105] While bacterial loadin tissue was slightly reduced the major change was ame-lioration of granulomatous severity It is noteworthy thatbovine lactoferrin reduced bacterial burden accompaniedby an increase in classical proinflammatory responses whiledecreasing overall lung immunopathology [105] Specificallylactoferrin-treated mice increased numbers of CD4+ IFN-120574+ and IL-17 producing cells in the lung It was shown thatlactoferrin by itself was not bactericidal but rather enhancedIFN-120574mediatedMTBkilling bymacrophages in a nitric oxidedependent manner These studies indicate that lactoferrinmay be a novel therapeutic for the treatment of tuberculosisand may be useful in infectious diseases to reduced immune-mediated tissue damage

Finally the molecular mechanism underlying resolutionof the granulomatous lesion is not yet defined However arole for maintaining strong Th1 responses could possibly beattributed to relationships due to COX-2 inhibition by lacto-ferrin in inducedmacrophages (WBCs) [106] and knowledgethat prostaglandin E2 (PGE2) is able to shift T helperresponses during BCG activation of innate macrophages[107] It had been previously demonstrated that the PGE2response could be reversed by a COX-2 inhibitor [108]Therefore lactoferrin functioning as a COX-2 inhibitor couldbe an indirect method to maintain proinflammatory activityof macrophages when infected

3 Conclusions

The studies here indicate that lactoferrin may be a usefuladjunct therapeutic for amelioration of pathological responsein an in vivo model of TB granulomas Of high importancethe change in pathology is accomplished with no loss ofinnate immune function which would be critical for defenseagainst pathogenic organisms A question arises concerningthe utility of using heterologous lactoferrins in the mousemodel Preliminary investigations show that a newly devel-oped recombinant mouse lactoferrin produced in CHO sta-ble cell lines functions in a manner very similar to the bovineand human recombinant to limit immunopathology afteroral delivery Mechanistically it was shown that the mouseand human forms are nearly identical in modulation of den-dritic cell function in response to Bacillus Calmette-Guerin(BCG) [25 109] The dendritic cell population respondswith increased inflammatory cytokines and a shift towardsMHC Class II expression when BCG is combined withthe lactoferrins However more recent investigations revealthat the macrophage population has a unique trend withpotentially decreased antigen presentation and subsequentT-cell stimulatory activity [110] Overall these results givegreat confidence to move forward to test novel recombinantlactoferrins for adjunct clinical therapeutic effects to alterimmunedependent granulomatous pathology occurring dur-ing tuberculosis infection in humansThe studies here woulddrive the next goals to explore novel recombinant lactoferrinsto mediate reductions in lung histopathology and bacterialburden in BSL3 in vivo models of MTB infection Further-more we hypothesize that reduction of TB-induced inflam-matory pathology would also allow enhanced penetration

Mediators of Inflammation 5

2000

1500

1000

500

0

TDM

Treatment

lowast lowast lowast lowast

IL-12

p40

conc

entr

atio

n (p

glu

ng)

Lung IL-12p40

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

6000

2000

3000

4000

5000

1000

0

TDM

Treatment

lowast lowast lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

Lung TNF-120572

IL-6

conc

entr

atio

n (p

glu

ng)

2000

1500

1000

500

0

TDM

Treatment

lowast lowast lowast

Lung IL-6

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

40000

20000

30000

10000

0

TDM

Treatment

lowast lowast lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

IL-1120573

conc

entr

atio

n (p

glu

ng)

Lung IL-1120573

IL-10

conc

entr

atio

n (p

glu

ng)

Lung IL-10300

200

100

0

TDM

Treatment

lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

TNF-120572

conc

entr

atio

n (p

glu

ng)

Figure 3 Proinflammatory cytokines in lungs are reduced in lactoferrin-treated mice after challenge with TDM BALBc mice werechallenged with 25120583g TDM in a water-in-oil emulsion Bovine or recombinant human lactoferrin (1 or 2mg) was orally administered at4 and 6 days after TDM challenge Methods of TDM induction are described elsewhere [38 101] Lungs were isolated on day 7 after TDMchallenge cytokines were assessed by ELISA values are shown per lung for individual mice lowast119901 lt 005 compared to the TDM emulsion alonetreated mice

of antimycobacterial drugs to sites harboring infectiousagents in essence permitting an efficient mechanism totreat TB infection Overall these studies permit confidenceto target clinical implications and test novel strategies to

modulate MTB-induced granulomatous responses to allowmore efficacious delivery of therapeutics to mycobacterialorganisms residing within lung tissue during activemycobac-terial disease states

6 Mediators of Inflammation

BALB

cC5

7BL

6TDM alone TDM + LFNaiumlve

TDM alone TDM + LFNaiumlve

Figure 4 Granuloma response to TDM challenge in oral recombinant human lactoferrin-treated mice Two strains of mice (BALBc andC57BL6) were challenged with 25120583g TDM in a water-in-oil emulsion as described [38 101] Human lactoferrin (recombinant 1mg) wasgiven orally at 4 and 6 days after TDM challenge Comparisons are made to naıve mice and to mice only administered TDM HampE staining100x Representative sections from mice at 7 days after TDM challenge119873 ge 8 per experimental group

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

This work was supported in part by NIH Grants 1R41-AI117990-01 and R42-AI051050-05The authors acknowledgePharmaReview Corp (Houston TX) for their kind gifts oflactoferrins and thank Marian L Kruzel PhD and Shen-An Hwang PhD for their insights into the research projectThe authors also thank Ron Gadot and Ai Yamazaki for theirtechnical contributions to the research presented

References

[1] P E M Fine ldquoHerd immunity history theory practicerdquo Epide-miologic Reviews vol 15 no 2 pp 265ndash302 1993

[2] L Ramakrishnan ldquoRevisiting the role of the granuloma intuberculosisrdquo Nature Reviews Immunology vol 12 no 5 pp352ndash366 2012

[3] M Silva Miranda A Breiman S Allain F Deknuydt andF Altare ldquoThe tuberculous granuloma an unsuccessful hostdefence mechanism providing a safety shelter for the bacteriardquoClinical amp Developmental Immunology vol 2012 Article ID139127 14 pages 2012

[4] S A Khader J Rangel-Moreno J J Fountain et al ldquoIn amurinetuberculosis model the absence of homeostatic chemokinesdelays granuloma formation and protective immunityrdquo Journalof Immunology vol 183 no 12 pp 8004ndash8014 2009

[5] AMCooper and S A Khader ldquoThe role of cytokines in the ini-tiation expansion and control of cellular immunity to tubercu-losisrdquo Immunological Reviews vol 226 no 1 pp 191ndash204 2008

[6] IMOrme and R J Basaraba ldquoThe formation of the granulomain tuberculosis infectionrdquo Seminars in Immunology vol 26 no6 pp 601ndash609 2014

[7] I M Orme R T Robinson and A M Cooper ldquoThe balancebetween protective and pathogenic immune responses in theTB-infected lungrdquo Nature Immunology vol 16 no 1 pp 57ndash632014

[8] J K Actor M Olsen C Jagannath and R L Hunter ldquoRela-tionship of survival organism containment and granulomaformation in acute murine tuberculosisrdquo Journal of Interferonand Cytokine Research vol 19 no 10 pp 1183ndash1193 1999

[9] J L Flynn and J Chan ldquoWhatrsquos good for the host is good for thebugrdquo Trends in Microbiology vol 13 no 3 pp 98ndash102 2005

[10] D G Russell ldquoMycobacterium tuberculosis and the intimatediscourse of a chronic infectionrdquo Immunological Reviews vol240 no 1 pp 252ndash268 2011

[11] R L Hunter ldquoPathology of post primary tuberculosis of thelung an illustrated critical reviewrdquo Tuberculosis vol 91 no 6pp 497ndash509 2011

[12] R L Hunter J K Actor S-A Hwang V Karev and C Jagan-nath ldquoPathogenesis of post primary tuberculosis immunity andhypersensitivity in the development of cavitiesrdquoAnnals of Clini-cal and Laboratory Science vol 44 no 4 pp 365ndash387 2014

[13] K J Welsh R L Hunter and J K Actor ldquoTrehalose 661015840-dimy-colatemdasha coat to regulate tuberculosis immunopathogenesisrdquoTuberculosis vol 93 supplement pp S3ndashS9 2013

[14] M-J Kim H C Wainwright M Locketz et al ldquoCaseation ofhuman tuberculosis granulomas correlates with elevated hostlipid metabolismrdquo EMBO Molecular Medicine vol 2 no 7 pp258ndash274 2010

[15] J K Actor E Breij R A Wetsel H Hoffmann R L HunterJr and C Jagannath ldquoA role for complement C5 in organ-ism containment and granulomatous response during murine

Mediators of Inflammation 7

tuberculosisrdquo Scandinavian Journal of Immunology vol 53 no5 pp 464ndash474 2001

[16] J K Actor C D Leonard V E Watson et al ldquoCytokinemRNAexpression and serumcortisol evaluation duringmurinelung inflammation induced by Mycobacterium tuberculosisrdquoCombinatorial Chemistry and High Throughput Screening vol3 no 4 pp 343ndash351 2000

[17] C W Borders A Courtney K Ronen et al ldquoRequisite role forcomplement C5 and the C5a receptor in granulomatous res-ponse to mycobacterial glycolipid trehalose 661015840-dimycolaterdquoScandinavian Journal of Immunology vol 62 no 2 pp 123ndash1302005

[18] R L Hunter C Jagannath and J K Actor ldquoPathology of post-primary tuberculosis in humans and mice contradiction oflong-held beliefsrdquo Tuberculosis vol 87 no 4 pp 267ndash278 2007

[19] C Jagannath H Hoffmann E Sepulveda J K Actor R AWetsel and R L Hunter Jr ldquoHypersusceptibility of AJ miceto tuberculosis is in part due to a deficiency of the fifth comple-ment component (C5)rdquo Scandinavian Journal of Immunologyvol 52 no 4 pp 369ndash379 2000

[20] V E Watson L L Hill L B Owen-Schaub et al ldquoApoptosis inMycobacterium tuberculosis infection in mice exhibiting variedimmunopathologyrdquo Journal of Pathology vol 190 no 2 pp 211ndash220 2000

[21] R H Copenhaver E Sepulveda L Y Armitige et al ldquoAmutantof Mycobacterium tuberculosis H37Rv that lacks expressionof antigen 85A is attenuated in mice but retains vaccinogenicpotentialrdquo Infection and Immunity vol 72 no 12 pp 7084ndash7095 2004

[22] S-A Hwang R Arora M L Kruzel and J K Actor ldquoLactofer-rin enhances efficacy of the BCG vaccine comparison betweentwo inbred mice strains (C57BL6 and BALBc)rdquo Tuberculosisvol 89 supplement 1 pp S49ndashS54 2009

[23] S-A Hwang K Wilk M L Kruzel and J K Actor ldquoA novelrecombinant human lactoferrin augments the BCG vaccine andprotects alveolar integrity upon infection with Mycobacteriumtuberculosis in micerdquo Vaccine vol 27 no 23 pp 3026ndash30342009

[24] S-A Hwang K M Wilk M Budnicka et al ldquoLactoferrinenhanced efficacy of the BCG vaccine to generate host protec-tive responses against challenge with virulent Mycobacteriumtuberculosisrdquo Vaccine vol 25 no 37-38 pp 6730ndash6743 2007

[25] S-A Hwang and J K Actor ldquoLactoferrin modulation of BCG-infected dendritic cell functionsrdquo International Immunologyvol 21 no 10 pp 1185ndash1197 2009

[26] S-A Hwang M L Kruzel and J K Actor ldquoLactoferrin aug-ments BCG vaccine efficacy to generate T helper response andsubsequent protection against challenge with virulentMycobac-terium tuberculosisrdquo International Immunopharmacology vol 5no 3 pp 591ndash599 2005

[27] K M Wilk S-A Hwang and J K Actor ldquoLactoferrin mod-ulation of antigen-presenting-cell response to BCG infectionrdquoPostepy Higieny i Medycyny Doswiadczalnej vol 61 pp 277ndash282 2007

[28] S A Hwang M L Kruzel and J K Actor ldquoCHO expressedrecombinant human lactoferrin as an adjuvant for BCGrdquoInternational Journal of Immunopathology and Pharmacologyvol 28 no 4 pp 452ndash468 2015

[29] R L Perez J Roman G W Staton Jr and R L HunterldquoExtravascular coagulation and fibrinolysis in murine lunginflammation induced by the mycobacterial cord factor

trehalose-66rsquo-dimycolaterdquoAmerican Journal of Respiratory andCritical Care Medicine vol 149 no 2 pp 510ndash518 1994

[30] C A Behling R L Perez M R Kidd G W Staton Jr and RL Hunter ldquoInduction of pulmonary granulomas macrophageprocoagulant activity and tumor necrosis factor-alpha by tre-halose glycolipidsrdquo Annals of Clinical and Laboratory Sciencevol 23 no 4 pp 256ndash266 1993

[31] G S Retzinger S C Meredith R L Hunter K Takayama andF J Kezdy ldquoIdentification of the physiologically active state ofthe mycobacterial glycolipid trehalose 661015840-dimycolate and therole of fibrinogen in the biologic activities of trehalose 661015840-dimycolate monolayersrdquo Journal of Immunology vol 129 no 2pp 735ndash744 1982

[32] G S Retzinger S C Meredith K Takayama R L Hunter andF J Kezdy ldquoThe role of surface in the biological activities oftrehalose 661015840-dimycolate Surface properties and developmentof amodel systemrdquoThe Journal of Biological Chemistry vol 256no 15 pp 8208ndash8216 1981

[33] R L Perez J Roman S Roser et al ldquoCytokinemessage and pro-tein expression during lung granuloma formation and resolu-tion induced by the mycobacterial cord factor trehalose-661015840-dimycolaterdquo Journal of Interferon amp Cytokine Research vol 20no 9 pp 795ndash804 2000

[34] R L Hunter M R Olsen C Jagannath and J K Actor ldquoMult-iple roles of cord factor in the pathogenesis of primary sec-ondary and cavitary tuberculosis including a revised descrip-tion of the pathology of secondary diseaserdquo Annals of Clinicaland Laboratory Science vol 36 no 4 pp 371ndash386 2006

[35] J K Actor ldquoMycobacterial TDM a coat to modulate post pri-mary pathogenesisrdquoMycobacterial Diseases vol 2 no 1 articlee106 2012

[36] R L HunterMOlsen C Jagannath and J K Actor ldquoTrehalose661015840-dimycolate and lipid in the pathogenesis of caseatinggranulomas of tuberculosis in micerdquo The American Journal ofPathology vol 168 no 4 pp 1249ndash1261 2006

[37] A N Abbott T V Guidry K J Welsh et al ldquo11Beta-hydroxy-steroid dehydrogenases are regulated during the pulmonarygranulomatous response to the mycobacterial glycolipidtrehalose-661015840-dimycolaterdquo NeuroImmunoModulation vol 16no 3 pp 147ndash154 2009

[38] K J Welsh A N Abbott S-A Hwang et al ldquoA role for tumournecrosis factor-120572 complement C5 and interleukin-6 in theinitiation and development of the mycobacterial cord factortrehalose 661015840-dimycolate induced granulomatous responserdquoMicrobiology vol 154 no 6 pp 1813ndash1824 2008

[39] A Bekierkunst and E Yarkoni ldquoGranulomatous hypersensitiv-ity to trehalose-66rsquo-dimycolate (cord factor) in mice infectedwith BCGrdquo Infection and Immunity vol 7 no 4 pp 631ndash6381973

[40] H Yamagami TMatsumoto N Fujiwara et al ldquoTrehalose 661015840-dimycolate (cord factor) ofMycobacterium tuberculosis inducesforeign-body- and hypersensitivity-type granulomas in micerdquoInfection and Immunity vol 69 no 2 pp 810ndash815 2001

[41] T VGuidry R LHunter Jr and J K Actor ldquoMycobacterial gly-colipid trehalose 661015840-dimycolate-induced hypersensitive gran-ulomas contribution of CD4+ lymphocytesrdquoMicrobiology vol153 no 10 pp 3360ndash3369 2007

[42] E Rhoades F-F Hsu J B Torrelles J Turk D Chatterjee andD G Russell ldquoIdentification and macrophage-activating activ-ity of glycolipids released from intracellular Mycobacteriumbovis BCGrdquoMolecular Microbiology vol 48 no 4 pp 875ndash8882003

8 Mediators of Inflammation

[43] MMHossain andM-N Norazmi ldquoPattern recognition recep-tors and cytokines in Mycobacterium tuberculosis infectionmdashthe double-edged swordrdquo BioMed Research International vol2013 Article ID 179174 18 pages 2013

[44] D M E Bowdish K Sakamoto M-J Kim et al ldquoMARCOTLR2 and CD14 are required for macrophage cytokine res-ponses to mycobacterial trehalose dimycolate and Mycobac-terium tuberculosisrdquo PLoS Pathogens vol 5 no 6 Article IDe1000474 2009

[45] E Mortaz I M Adcock P Tabarsi et al ldquoInteraction of patternrecognition receptors withMycobacterium tuberculosisrdquo Journalof Clinical Immunology vol 35 no 1 pp 1ndash10 2015

[46] D G Russell ldquoWho puts the tubercle in tuberculosisrdquo NatureReviews Microbiology vol 5 no 1 pp 39ndash47 2007

[47] H Schoenen B Bodendorfer K Hitchens et al ldquoCutting Edgemincle is essential for recognition and adjuvanticity of themycobacterial cord factor and its synthetic analog trehalose-dibehenaterdquo Journal of Immunology vol 184 no 6 pp 2756ndash2760 2010

[48] E Ishikawa T Ishikawa Y S Morita et al ldquoDirect recognitionof themycobacterial glycolipid trehalose dimycolate by C-typelectinMinclerdquo Journal of ExperimentalMedicine vol 206 no 13pp 2879ndash2888 2009

[49] I Matsunaga and D B Moody ldquoMincle is a long soughtreceptor for mycobacterial cord factorrdquo Journal of ExperimentalMedicine vol 206 no 13 pp 2865ndash2868 2009

[50] K Werninghaus A Babiak O Gross et al ldquoAdjuvanticity ofa synthetic cord factor analogue for subunit Mycobacteriumtuberculosis vaccination requires FclR120574-Syk-Card9-dependentinnate immune activationrdquo Journal of Experimental Medicinevol 206 no 1 pp 89ndash97 2009

[51] S LeibundGut-LandmannOGrossM J Robinson et al ldquoSyk-and CARD9-dependent coupling of innate immunity to theinduction of T helper cells that produce interleukin 17rdquo NatureImmunology vol 8 no 6 pp 630ndash638 2007

[52] J Indrigo R L Hunter Jr and J K Actor ldquoInfluence of tre-halose 661015840-dimycolate (TDM) during mycobacterial infectionof bone marrow macrophagesrdquoMicrobiology vol 148 no 7 pp1991ndash1998 2002

[53] J Indrigo R L Hunter Jr and J K Actor ldquoCord factortrehalose 661015840-dimycolate (TDM) mediates trafficking eventsduringmycobacterial infection ofmurinemacrophagesrdquoMicro-biology vol 149 no 8 pp 2049ndash2059 2003

[54] J K Actor M Olsen R L Hunter Jr and Y-J Geng ldquoDys-regulated response to mycobacterial cord factor trehalose-661015840-dimycolate inCD1Dminusminusmicerdquo Journal of Interferon andCytokineResearch vol 21 no 12 pp 1089ndash1096 2001

[55] J K Actor J Indrigo C M Beachdel et al ldquoMycobacterial gly-colipid cord factor trehalose 661015840-dimycolate causes a decreasein serum cortisol during the granulomatous responserdquo Neuro-ImmunoModulation vol 10 no 5 pp 270ndash282 2002

[56] A N Abbott K J Welsh S-A Hwang et al ldquoIL-6 mediates11120573hsd type 2 to effect progression of the mycobacterial cordfactor trehalose 661015840-dimycolate-induced granulomatous res-ponserdquo NeuroImmunoModulation vol 18 no 4 pp 212ndash2252011

[57] T V Guidry M Olsen K-S Kil R L Hunter Jr Y-J Gengand J K Actor ldquoFailure of CD1Dminusminusmice to elicit hypersensitivegranulomas to mycobacterial cord factor trehalose 661015840-dimy-colaterdquo Journal of Interferon amp Cytokine Research vol 24 no 6pp 362ndash371 2004

[58] TVGuidry R LHunter Jr and J KActor ldquoCD3+ cells transferthe hypersensitive granulomatous response to mycobacterialglycolipid trehalose 661015840-dimycolate in micerdquoMicrobiology vol152 no 12 pp 3765ndash3775 2006

[59] B Lonnerdal and S Iyer ldquoLactoferrin molecular structure andbiological functionrdquoAnnual Review of Nutrition vol 15 pp 93ndash110 1995

[60] L Sanchez M Calvo and J H Brock ldquoBiological role of lacto-ferrinrdquo Archives of Disease in Childhood vol 67 no 5 pp 657ndash661 1992

[61] P Manzoni M Rinaldi S Cattani et al ldquoBovine lactoferrinsupplementation for prevention of late-onset sepsis in verylow-birth-weight neonates a randomized trialrdquo Journal of theAmerican Medical Association vol 302 no 13 pp 1421ndash14282009

[62] S Baveye E Elass J Mazurier G Spik and D Legrand ldquoLacto-ferrin a multifunctional glycoprotein involved in the modula-tion of the inflammatory processrdquoClinical Chemistry and Labo-ratory Medicine vol 37 no 3 pp 281ndash286 1999

[63] R D Baynes and W R Bezwoda ldquoLactoferrin and the inflam-matory responserdquo Advances in Experimental Medicine andBiology vol 357 pp 133ndash141 1994

[64] M Kocieba M Zimecki M Kruzel and J Actor ldquoThe adjuvantactivity of lactoferrin in the generation of DTH to ovalbumincan be inhibited by bovine serum albumin bearing alpha-D-mannopyranosyl residuesrdquoCellularampMolecular Biology Lettersvol 7 no 4 pp 1131ndash1136 2002

[65] C G Turin A Zea-Vera A Pezo et al ldquoLactoferrin for pre-vention of neonatal sepsisrdquo Biometals vol 27 no 5 pp 1007ndash1016 2014

[66] T Siqueiros-Cendon S Arevalo-Gallegos B F Iglesias-Figueroa I A Garcıa-Montoya J Salazar-Martınez and QRascon-Cruz ldquoImmunomodulatory effects of lactoferrinrdquo ActaPharmacologica Sinica vol 35 no 5 pp 557ndash566 2014

[67] D Latorre F Berlutti P Valenti S Gessani and P Puddu ldquoLFimmunomodulatory strategies mastering bacterial endotoxinrdquoBiochemistry and Cell Biology vol 90 no 3 pp 269ndash278 2012

[68] D Legrand ldquoLactoferrin a key molecule in immune andinflammatory processesrdquo Biochemistry and Cell Biology vol 90no 3 pp 252ndash268 2012

[69] S-A Hwang K M Wilk Y A Bangale M L Kruzel and JK Actor ldquoLactoferrin modulation of IL-12 and IL-10 responsefrom activated murine leukocytesrdquo Medical Microbiology andImmunology vol 196 no 3 pp 171ndash180 2007

[70] M Zimecki J Mazurier G Spik and J A Kapp ldquoHuman lacto-ferrin induces phenotypic and functional changes in murinesplenic B cellsrdquo Immunology vol 86 no 1 pp 122ndash127 1995

[71] M Zimecki J Artym G Chodaczek et al ldquoImmunoregulatoryfunction of lactoferrin in immunosuppressed and autoimmuneanimalsrdquo Postepy Higieny i Medycyny Doswiadczalnej vol 61pp 283ndash287 2007

[72] M Zimecki and M L Kruzel ldquoMilk-derived proteins andpeptides of potential therapeutic and nutritive valuerdquo Journal ofExperimental Therapeutics and Oncology vol 6 no 2 pp 89ndash106 2007

[73] M Zimecki RMiędzybrodzki J Mazurier andG Spik ldquoRegu-latory effects of lactoferrin and lipopolysaccharide on LFA-1 expression on human peripheral blood mononuclear cellsrdquoArchivum Immunologiae et Therapiae Experimentalis vol 47no 4 pp 257ndash264 1999

Mediators of Inflammation 9

[74] M L Kruzel and M Zimecki ldquoLactoferrin and immunologicdissonance clinical implicationsrdquo Archivum Immunologiae etTherapiae Experimentalis vol 50 no 6 pp 399ndash410 2002

[75] W I Weis M E Taylor and K Drickamer ldquoThe C-type lectinsuperfamily in the immune systemrdquo Immunological Reviewsvol 163 pp 19ndash34 1998

[76] W I Weis K Drickamer and W A Hendrickson ldquoStructureof a C-type mannose-binding protein complexed with an oli-gosacchariderdquo Nature vol 360 no 6400 pp 127ndash134 1992

[77] T Zagulski Z Jarzabek A Zagulska M Jaszczak I EKochanowska and M Zimecki ldquoLactoferrin stimulates killingand clearance of bacteria but does not prevent mortality of dia-betic micerdquo Archivum Immunologiae et Therapiae Experimen-talis vol 49 no 6 pp 431ndash438 2001

[78] M Kruzel Y Harai C Chen and A Castro ldquoRole of lactoferrinin development of systemic inflammatory response syndrome(SIRS)rdquo in Proceedings of the 2nd International ConferenceProgress in Intensive Care Medicine pp 11ndash12 Wroclaw Poland1998

[79] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoThe guta key metabolic organ protected by lactoferrin during experi-mental systemic inflammation in micerdquo in Advances in Lacto-ferrin Research vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Springer New York NY USA 1998

[80] MKruzel T Zagulski andMZimecki ldquoLactoferrin and insult-induced metabolic imbalance in humans and other animalsrdquoin Lactoferrin Structure Function and Applications Proceedingsof the 4th International Conference on Lactoferrin StructureFunction and Applications held in Sapporo Japan 18ndash22 May1999 pp 301ndash311 ICS Elsevier New York NY USA 2000

[81] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoLactofer-rin protects gut mucosal integrity during endotoxemia inducedby lipopolysaccharide in micerdquo Inflammation vol 24 no 1 pp33ndash44 2000

[82] E Zaczynska M Kocięba E Sliwinska and M ZimeckildquoBovine lactoferrin enhances proliferation of human peripheralblood lymphocytes and induces cytokine production in wholeblood culturesrdquo Advances in Clinical and Experimental Medi-cine vol 23 pp 871ndash876 2014

[83] Z Wei T Nishimura and S Yoshida ldquoCharacterization ofglycans in a lactoferrin isoform lactoferrin-ardquo Journal of DairyScience vol 84 no 12 pp 2584ndash2590 2001

[84] B-K Choi J K Actor S Rios et al ldquoRecombinant humanlactoferrin expressed in glycoengineered Pichia pastoris effectof terminal N-acetylneuraminic acid on in vitro secondaryhumoral immune responserdquoGlycoconjugate Journal vol 25 no6 pp 581ndash593 2008

[85] B Samyn-Petit J-PWajdaDubos F Chirat et al ldquoComparativeanalysis of the site-specific N-glycosylation of human lactofer-rin produced in maize and tobacco plantsrdquo European Journal ofBiochemistry vol 270 no 15 pp 3235ndash3242 2003

[86] M Zimecki J Artym M Kocięba M Duk and M L KruzelldquoThe effect of carbohydrate moiety structure on the immu-noregulatory activity of lactoferrin in vitrordquo Cellular ampMolecu-lar Biology Letters vol 19 no 2 pp 284ndash296 2014

[87] J K Actor S-A Hwang and M L Kruzel ldquoLactoferrin as anatural immune modulatorrdquo Current Pharmaceutical Designvol 15 no 17 pp 1956ndash1973 2009

[88] M L Kruzel J K Actor I Boldogh and M ZimeckildquoLactoferrin in health and diseaserdquo Postepy Higieny i MedycynyDoswiadczalnej vol 61 pp 261ndash267 2007

[89] S Baveye E Elass D G Fernig C Blanquart J Mazurier andD Legrand ldquoHuman lactoferrin interacts with soluble CD14and inhibits expression of endothelial adhesion molecules E-selectin and ICAM-1 induced by the CD14-lipopolysaccharidecomplexrdquo Infection and Immunity vol 68 no 12 pp 6519ndash65252000

[90] C S Curran K P Demick and J M Mansfield ldquoLactoferrinactivates macrophages via TLR4-dependent and -independentsignaling pathwaysrdquo Cellular Immunology vol 242 no 1 pp23ndash30 2006

[91] P Puddu D Latorre M Carollo et al ldquoBovine lactoferrincounteracts Toll-Like receptor mediated activation signals inantigen presenting cellsrdquo PLoS ONE vol 6 no 7 Article IDe22504 2011

[92] T M Doherty and M Arditi ldquoTB or not TB That is the ques-tionmdashdoes TLR signaling hold the answerrdquoThe Journal of Clin-ical Investigation vol 114 no 12 pp 1699ndash1703 2004

[93] M A Rahman P Sobia N Gupta L Van Kaer and GDas ldquoMycobacterium tuberculosis subverts the TLR-2mdashMyD88pathway to facilitate its translocation into the cytosolrdquo PLoSONE vol 9 no 1 Article ID e86886 2014

[94] J ArtymM Zimecki andM L Kruzel ldquoEnhanced clearance ofEscherichia coli and Staphylococcus aureus in mice treated withcyclophosphamide and lactoferrinrdquo International Immunophar-macology vol 4 no 9 pp 1149ndash1157 2004

[95] G Chodaczek A Saavedra-Molina A Bacsi M L Kruzel SSur and I Boldogh ldquoIron-mediated dismutation of superoxideanion augments antigen-induced allergic inflammation effectof lactoferrinrdquo Postepy Higieny i Medycyny Doswiadczalnej vol61 pp 268ndash276 2007

[96] M Kruzel Y Harai C Chen and A Castro ldquoThe Gut a keymetabolic organ protected by lactoferrin during experimentalsystemic inflammation in micerdquo in Advances in LactoferrinResearch G Spik D Legrand J Mazurier A Pierce and J-PPerraudin Eds vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Plenum Press 1998

[97] M L Kruzel A Bacsi B Choudhury S Sur and I BoldoghldquoLactoferrin decreases pollen antigen-induced allergic airwayinflammation in a murine model of asthmardquo Immunology vol119 no 2 pp 159ndash166 2006

[98] J K Actor S-A Hwang M Olsen M Zimecki R L HunterJr and M L Kruzel ldquoLactoferrin immunomodulation of DTHresponse in micerdquo International Immunopharmacology vol 2no 4 pp 475ndash486 2002

[99] S-A Hwang M L Kruzel and J K Actor ldquoInfluence of bovinelactoferrin on expression of presentation molecules on BCG-infected bonemarrow derivedmacrophagesrdquo Biochimie vol 91no 1 pp 76ndash85 2009

[100] M Zimecki J Mazurier M Machnicki Z Wieczorek JMontreuil and G Spik ldquoImmunostimulatory activity of lacto-transferrin andmaturation of CD4minus CD8minusmurine thymocytesrdquoImmunology Letters vol 30 no 1 pp 119ndash123 1991

[101] K J Welsh S-A Hwang R L Hunter M L Kruzel and JK Actor ldquoLactoferrin modulation of mycobacterial cord factortrehalose 6-6rsquo-dimycolate induced granulomatous responserdquoTranslational Research vol 156 no 4 pp 207ndash215 2010

[102] M L Bourigault N Segueni S Rose et al ldquoRelative contri-bution of IL-1alpha IL-1beta and TNF to the host responseto Mycobacterium tuberculosis and attenuated M bovis BCGrdquoImmunity Inflammation and Disease vol 1 no 1 pp 47ndash622013

10 Mediators of Inflammation

[103] C Holscher R A Atkinson B Arendse et al ldquoA protective andagonistic function of IL-12p40 in mycobacterial infectionrdquoTheJournal of Immunology vol 167 no 12 pp 6957ndash6966 2001

[104] M Jacobs L Fick N Allie N Brown and B Ryffel ldquoEnhancedimmune response in Mycobacterium bovis bacille calmetteguerin (BCG)-infected IL-10-deficientmicerdquoClinical Chemistryand Laboratory Medicine vol 40 no 9 pp 893ndash902 2002

[105] K J Welsh S-A Hwang S Boyd M L Kruzel R L Hunterand J K Actor ldquoInfluence of oral lactoferrin onMycobacteriumtuberculosis induced immunopathologyrdquo Tuberculosis vol 91supplement 1 pp S105ndashS113 2011

[106] M L Kruzel J K Actor M Zimecki et al ldquoNovel recombinanthuman lactoferrin differential activation of oxidative stressrelated gene expressionrdquo Journal of Biotechnology vol 168 no4 pp 666ndash675 2013

[107] Y Shibata J Gabbard M Yamashita et al ldquoHeat-killedBCG induces biphasic cyclooxygenase 2+ splenic macrophageformationmdashrole of IL-10 and bone marrow precursorsrdquo Journalof Leukocyte Biology vol 80 no 3 pp 590ndash598 2006

[108] H Yamada E Kuroda S Matsumoto T Matsumoto TYamada and U Yamashita ldquoProstaglandin E

2down-regulates

viable Bacille Calmette-Guerin-induced macrophage cytotoxi-city against murine bladder cancer cell MBT-2 in vitrordquoClinicaland Experimental Immunology vol 128 no 1 pp 52ndash58 2002

[109] S Hwang M L Kruzel and J K Actor ldquoEffects of CHO-expressed recombinant lactoferrins on mouse dendritic cellpresentation and functionrdquo Innate Immunity vol 21 no 5 pp553ndash561 2015

[110] K M OShea S-A Hwang and J K Actor ldquoImmune activity ofbcg infected mouse macrophages treated with a novel recom-binant mouse lactoferrinrdquo Annals of Clinical Lab Science vol45 no 5 pp 403ndash410 2015

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

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PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

4 Mediators of Inflammation

and adaptive events are not mutually exclusive but rathercomplementary and lend credence to utility of lactoferrin forimmune homeostasis in insult-inducedmetabolic disparities

22 Lactoferrin Modulation of the Tuberculoid GranulomaAn examination of lactoferrin to modulate granulomatousresponses was performed in mice injected intravenouslywith TDM A subset of mice were intravenously given 1mgbovine lactoferrin 24 h after TDM challenge Lung tissue wasanalyzed for histological response and for the productionof proinflammatory mediators Treatment with lactoferrinshowed statistically significant fewer and smaller granulomascompared to TDM alone [101] Of importance the protectiveproinflammatory mediators were not statistically diminishedby lactoferrin treatment Nontreated mice demonstrateda granuloma formation that correlated with an increasedproduction of tumor necrosis factor-120572 (TNF-120572) interleukin-(IL-) 1120573 and IL-6This is important as it suggests that innatefunctions critical during infectious disease are maintainedeven though immunopathology is reduced by treatment

In a similar manner the utility of oral delivered lactofer-rin was also examined As previously mentioned mice givenTDM alone also showed marked and statistically significantincreased production of proinflammatory cytokines TNF-120572IL-6 IL-1120573 IL-12p40 and KC (keratinocyte chemoattrac-tant) Oral treatment with recombinant human LF (producedin CHO cells) at 1 or 2mg led to statistically significantreduction (119901 lt 005) in cytokine levels at 7 days after TDMadministration in BALBc and C57BL6 mice (Figure 3 onlyBALBc data shown) This is consistent with reports thatthese proinflammatory cytokines are protective in similarinfectious challenge models [102 103] while molecules suchas IL-10 which are limited after lactoferrin administrationare detrimental during infection [104] In a similar mannerbovine lactoferrin given orally at 1 or 2mg doses also showedstatistical reduction in most cytokines although there wasgreater consistency of response in BALBc mice than thatfor the C57BL6 strain In particular the 1mg dosage ofrecombinant LF showed the most consistency in its ability tocontrol (reduce) production of proinflammatory cytokines

Of major interest the oral delivered human recombinantlactoferrin demonstrated biologically relevant reduction inpathology (Figure 4) similar to that seen in the publishedresults using intravenous administered bovine lactoferrin Infact both the bovine and the recombinant human lactoferrinswere able to reduce inflammation due toTDMchallenge withdoses of 1 or 2mg given on days 4 and 6 after challenge result-ing in clear reduction in granuloma size and frequency Themost effective dose for the recombinant human lactoferrinwas seen when used at 1mg with some efficacy also seengiven as a single dose only on day 4 (data not shown)

Another point of special interest is that the classicalcytokines critical for control of MTB namely TNF-120572 IL-6 and IFN-120574 were not statistically altered by lactoferrintreatment [105] This gave confidence that treatment wouldretain control of organisms after infectious challenge Indeedthis is what was seen in a similar experiment performedusing bovine lactoferrin administered in drinking water(5mgmL) given to mice aerosol infected with virulent

mycobacteria (Erdman strain) [105] While bacterial loadin tissue was slightly reduced the major change was ame-lioration of granulomatous severity It is noteworthy thatbovine lactoferrin reduced bacterial burden accompaniedby an increase in classical proinflammatory responses whiledecreasing overall lung immunopathology [105] Specificallylactoferrin-treated mice increased numbers of CD4+ IFN-120574+ and IL-17 producing cells in the lung It was shown thatlactoferrin by itself was not bactericidal but rather enhancedIFN-120574mediatedMTBkilling bymacrophages in a nitric oxidedependent manner These studies indicate that lactoferrinmay be a novel therapeutic for the treatment of tuberculosisand may be useful in infectious diseases to reduced immune-mediated tissue damage

Finally the molecular mechanism underlying resolutionof the granulomatous lesion is not yet defined However arole for maintaining strong Th1 responses could possibly beattributed to relationships due to COX-2 inhibition by lacto-ferrin in inducedmacrophages (WBCs) [106] and knowledgethat prostaglandin E2 (PGE2) is able to shift T helperresponses during BCG activation of innate macrophages[107] It had been previously demonstrated that the PGE2response could be reversed by a COX-2 inhibitor [108]Therefore lactoferrin functioning as a COX-2 inhibitor couldbe an indirect method to maintain proinflammatory activityof macrophages when infected

3 Conclusions

The studies here indicate that lactoferrin may be a usefuladjunct therapeutic for amelioration of pathological responsein an in vivo model of TB granulomas Of high importancethe change in pathology is accomplished with no loss ofinnate immune function which would be critical for defenseagainst pathogenic organisms A question arises concerningthe utility of using heterologous lactoferrins in the mousemodel Preliminary investigations show that a newly devel-oped recombinant mouse lactoferrin produced in CHO sta-ble cell lines functions in a manner very similar to the bovineand human recombinant to limit immunopathology afteroral delivery Mechanistically it was shown that the mouseand human forms are nearly identical in modulation of den-dritic cell function in response to Bacillus Calmette-Guerin(BCG) [25 109] The dendritic cell population respondswith increased inflammatory cytokines and a shift towardsMHC Class II expression when BCG is combined withthe lactoferrins However more recent investigations revealthat the macrophage population has a unique trend withpotentially decreased antigen presentation and subsequentT-cell stimulatory activity [110] Overall these results givegreat confidence to move forward to test novel recombinantlactoferrins for adjunct clinical therapeutic effects to alterimmunedependent granulomatous pathology occurring dur-ing tuberculosis infection in humansThe studies here woulddrive the next goals to explore novel recombinant lactoferrinsto mediate reductions in lung histopathology and bacterialburden in BSL3 in vivo models of MTB infection Further-more we hypothesize that reduction of TB-induced inflam-matory pathology would also allow enhanced penetration

Mediators of Inflammation 5

2000

1500

1000

500

0

TDM

Treatment

lowast lowast lowast lowast

IL-12

p40

conc

entr

atio

n (p

glu

ng)

Lung IL-12p40

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

6000

2000

3000

4000

5000

1000

0

TDM

Treatment

lowast lowast lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

Lung TNF-120572

IL-6

conc

entr

atio

n (p

glu

ng)

2000

1500

1000

500

0

TDM

Treatment

lowast lowast lowast

Lung IL-6

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

40000

20000

30000

10000

0

TDM

Treatment

lowast lowast lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

IL-1120573

conc

entr

atio

n (p

glu

ng)

Lung IL-1120573

IL-10

conc

entr

atio

n (p

glu

ng)

Lung IL-10300

200

100

0

TDM

Treatment

lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

TNF-120572

conc

entr

atio

n (p

glu

ng)

Figure 3 Proinflammatory cytokines in lungs are reduced in lactoferrin-treated mice after challenge with TDM BALBc mice werechallenged with 25120583g TDM in a water-in-oil emulsion Bovine or recombinant human lactoferrin (1 or 2mg) was orally administered at4 and 6 days after TDM challenge Methods of TDM induction are described elsewhere [38 101] Lungs were isolated on day 7 after TDMchallenge cytokines were assessed by ELISA values are shown per lung for individual mice lowast119901 lt 005 compared to the TDM emulsion alonetreated mice

of antimycobacterial drugs to sites harboring infectiousagents in essence permitting an efficient mechanism totreat TB infection Overall these studies permit confidenceto target clinical implications and test novel strategies to

modulate MTB-induced granulomatous responses to allowmore efficacious delivery of therapeutics to mycobacterialorganisms residing within lung tissue during activemycobac-terial disease states

6 Mediators of Inflammation

BALB

cC5

7BL

6TDM alone TDM + LFNaiumlve

TDM alone TDM + LFNaiumlve

Figure 4 Granuloma response to TDM challenge in oral recombinant human lactoferrin-treated mice Two strains of mice (BALBc andC57BL6) were challenged with 25120583g TDM in a water-in-oil emulsion as described [38 101] Human lactoferrin (recombinant 1mg) wasgiven orally at 4 and 6 days after TDM challenge Comparisons are made to naıve mice and to mice only administered TDM HampE staining100x Representative sections from mice at 7 days after TDM challenge119873 ge 8 per experimental group

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

This work was supported in part by NIH Grants 1R41-AI117990-01 and R42-AI051050-05The authors acknowledgePharmaReview Corp (Houston TX) for their kind gifts oflactoferrins and thank Marian L Kruzel PhD and Shen-An Hwang PhD for their insights into the research projectThe authors also thank Ron Gadot and Ai Yamazaki for theirtechnical contributions to the research presented

References

[1] P E M Fine ldquoHerd immunity history theory practicerdquo Epide-miologic Reviews vol 15 no 2 pp 265ndash302 1993

[2] L Ramakrishnan ldquoRevisiting the role of the granuloma intuberculosisrdquo Nature Reviews Immunology vol 12 no 5 pp352ndash366 2012

[3] M Silva Miranda A Breiman S Allain F Deknuydt andF Altare ldquoThe tuberculous granuloma an unsuccessful hostdefence mechanism providing a safety shelter for the bacteriardquoClinical amp Developmental Immunology vol 2012 Article ID139127 14 pages 2012

[4] S A Khader J Rangel-Moreno J J Fountain et al ldquoIn amurinetuberculosis model the absence of homeostatic chemokinesdelays granuloma formation and protective immunityrdquo Journalof Immunology vol 183 no 12 pp 8004ndash8014 2009

[5] AMCooper and S A Khader ldquoThe role of cytokines in the ini-tiation expansion and control of cellular immunity to tubercu-losisrdquo Immunological Reviews vol 226 no 1 pp 191ndash204 2008

[6] IMOrme and R J Basaraba ldquoThe formation of the granulomain tuberculosis infectionrdquo Seminars in Immunology vol 26 no6 pp 601ndash609 2014

[7] I M Orme R T Robinson and A M Cooper ldquoThe balancebetween protective and pathogenic immune responses in theTB-infected lungrdquo Nature Immunology vol 16 no 1 pp 57ndash632014

[8] J K Actor M Olsen C Jagannath and R L Hunter ldquoRela-tionship of survival organism containment and granulomaformation in acute murine tuberculosisrdquo Journal of Interferonand Cytokine Research vol 19 no 10 pp 1183ndash1193 1999

[9] J L Flynn and J Chan ldquoWhatrsquos good for the host is good for thebugrdquo Trends in Microbiology vol 13 no 3 pp 98ndash102 2005

[10] D G Russell ldquoMycobacterium tuberculosis and the intimatediscourse of a chronic infectionrdquo Immunological Reviews vol240 no 1 pp 252ndash268 2011

[11] R L Hunter ldquoPathology of post primary tuberculosis of thelung an illustrated critical reviewrdquo Tuberculosis vol 91 no 6pp 497ndash509 2011

[12] R L Hunter J K Actor S-A Hwang V Karev and C Jagan-nath ldquoPathogenesis of post primary tuberculosis immunity andhypersensitivity in the development of cavitiesrdquoAnnals of Clini-cal and Laboratory Science vol 44 no 4 pp 365ndash387 2014

[13] K J Welsh R L Hunter and J K Actor ldquoTrehalose 661015840-dimy-colatemdasha coat to regulate tuberculosis immunopathogenesisrdquoTuberculosis vol 93 supplement pp S3ndashS9 2013

[14] M-J Kim H C Wainwright M Locketz et al ldquoCaseation ofhuman tuberculosis granulomas correlates with elevated hostlipid metabolismrdquo EMBO Molecular Medicine vol 2 no 7 pp258ndash274 2010

[15] J K Actor E Breij R A Wetsel H Hoffmann R L HunterJr and C Jagannath ldquoA role for complement C5 in organ-ism containment and granulomatous response during murine

Mediators of Inflammation 7

tuberculosisrdquo Scandinavian Journal of Immunology vol 53 no5 pp 464ndash474 2001

[16] J K Actor C D Leonard V E Watson et al ldquoCytokinemRNAexpression and serumcortisol evaluation duringmurinelung inflammation induced by Mycobacterium tuberculosisrdquoCombinatorial Chemistry and High Throughput Screening vol3 no 4 pp 343ndash351 2000

[17] C W Borders A Courtney K Ronen et al ldquoRequisite role forcomplement C5 and the C5a receptor in granulomatous res-ponse to mycobacterial glycolipid trehalose 661015840-dimycolaterdquoScandinavian Journal of Immunology vol 62 no 2 pp 123ndash1302005

[18] R L Hunter C Jagannath and J K Actor ldquoPathology of post-primary tuberculosis in humans and mice contradiction oflong-held beliefsrdquo Tuberculosis vol 87 no 4 pp 267ndash278 2007

[19] C Jagannath H Hoffmann E Sepulveda J K Actor R AWetsel and R L Hunter Jr ldquoHypersusceptibility of AJ miceto tuberculosis is in part due to a deficiency of the fifth comple-ment component (C5)rdquo Scandinavian Journal of Immunologyvol 52 no 4 pp 369ndash379 2000

[20] V E Watson L L Hill L B Owen-Schaub et al ldquoApoptosis inMycobacterium tuberculosis infection in mice exhibiting variedimmunopathologyrdquo Journal of Pathology vol 190 no 2 pp 211ndash220 2000

[21] R H Copenhaver E Sepulveda L Y Armitige et al ldquoAmutantof Mycobacterium tuberculosis H37Rv that lacks expressionof antigen 85A is attenuated in mice but retains vaccinogenicpotentialrdquo Infection and Immunity vol 72 no 12 pp 7084ndash7095 2004

[22] S-A Hwang R Arora M L Kruzel and J K Actor ldquoLactofer-rin enhances efficacy of the BCG vaccine comparison betweentwo inbred mice strains (C57BL6 and BALBc)rdquo Tuberculosisvol 89 supplement 1 pp S49ndashS54 2009

[23] S-A Hwang K Wilk M L Kruzel and J K Actor ldquoA novelrecombinant human lactoferrin augments the BCG vaccine andprotects alveolar integrity upon infection with Mycobacteriumtuberculosis in micerdquo Vaccine vol 27 no 23 pp 3026ndash30342009

[24] S-A Hwang K M Wilk M Budnicka et al ldquoLactoferrinenhanced efficacy of the BCG vaccine to generate host protec-tive responses against challenge with virulent Mycobacteriumtuberculosisrdquo Vaccine vol 25 no 37-38 pp 6730ndash6743 2007

[25] S-A Hwang and J K Actor ldquoLactoferrin modulation of BCG-infected dendritic cell functionsrdquo International Immunologyvol 21 no 10 pp 1185ndash1197 2009

[26] S-A Hwang M L Kruzel and J K Actor ldquoLactoferrin aug-ments BCG vaccine efficacy to generate T helper response andsubsequent protection against challenge with virulentMycobac-terium tuberculosisrdquo International Immunopharmacology vol 5no 3 pp 591ndash599 2005

[27] K M Wilk S-A Hwang and J K Actor ldquoLactoferrin mod-ulation of antigen-presenting-cell response to BCG infectionrdquoPostepy Higieny i Medycyny Doswiadczalnej vol 61 pp 277ndash282 2007

[28] S A Hwang M L Kruzel and J K Actor ldquoCHO expressedrecombinant human lactoferrin as an adjuvant for BCGrdquoInternational Journal of Immunopathology and Pharmacologyvol 28 no 4 pp 452ndash468 2015

[29] R L Perez J Roman G W Staton Jr and R L HunterldquoExtravascular coagulation and fibrinolysis in murine lunginflammation induced by the mycobacterial cord factor

trehalose-66rsquo-dimycolaterdquoAmerican Journal of Respiratory andCritical Care Medicine vol 149 no 2 pp 510ndash518 1994

[30] C A Behling R L Perez M R Kidd G W Staton Jr and RL Hunter ldquoInduction of pulmonary granulomas macrophageprocoagulant activity and tumor necrosis factor-alpha by tre-halose glycolipidsrdquo Annals of Clinical and Laboratory Sciencevol 23 no 4 pp 256ndash266 1993

[31] G S Retzinger S C Meredith R L Hunter K Takayama andF J Kezdy ldquoIdentification of the physiologically active state ofthe mycobacterial glycolipid trehalose 661015840-dimycolate and therole of fibrinogen in the biologic activities of trehalose 661015840-dimycolate monolayersrdquo Journal of Immunology vol 129 no 2pp 735ndash744 1982

[32] G S Retzinger S C Meredith K Takayama R L Hunter andF J Kezdy ldquoThe role of surface in the biological activities oftrehalose 661015840-dimycolate Surface properties and developmentof amodel systemrdquoThe Journal of Biological Chemistry vol 256no 15 pp 8208ndash8216 1981

[33] R L Perez J Roman S Roser et al ldquoCytokinemessage and pro-tein expression during lung granuloma formation and resolu-tion induced by the mycobacterial cord factor trehalose-661015840-dimycolaterdquo Journal of Interferon amp Cytokine Research vol 20no 9 pp 795ndash804 2000

[34] R L Hunter M R Olsen C Jagannath and J K Actor ldquoMult-iple roles of cord factor in the pathogenesis of primary sec-ondary and cavitary tuberculosis including a revised descrip-tion of the pathology of secondary diseaserdquo Annals of Clinicaland Laboratory Science vol 36 no 4 pp 371ndash386 2006

[35] J K Actor ldquoMycobacterial TDM a coat to modulate post pri-mary pathogenesisrdquoMycobacterial Diseases vol 2 no 1 articlee106 2012

[36] R L HunterMOlsen C Jagannath and J K Actor ldquoTrehalose661015840-dimycolate and lipid in the pathogenesis of caseatinggranulomas of tuberculosis in micerdquo The American Journal ofPathology vol 168 no 4 pp 1249ndash1261 2006

[37] A N Abbott T V Guidry K J Welsh et al ldquo11Beta-hydroxy-steroid dehydrogenases are regulated during the pulmonarygranulomatous response to the mycobacterial glycolipidtrehalose-661015840-dimycolaterdquo NeuroImmunoModulation vol 16no 3 pp 147ndash154 2009

[38] K J Welsh A N Abbott S-A Hwang et al ldquoA role for tumournecrosis factor-120572 complement C5 and interleukin-6 in theinitiation and development of the mycobacterial cord factortrehalose 661015840-dimycolate induced granulomatous responserdquoMicrobiology vol 154 no 6 pp 1813ndash1824 2008

[39] A Bekierkunst and E Yarkoni ldquoGranulomatous hypersensitiv-ity to trehalose-66rsquo-dimycolate (cord factor) in mice infectedwith BCGrdquo Infection and Immunity vol 7 no 4 pp 631ndash6381973

[40] H Yamagami TMatsumoto N Fujiwara et al ldquoTrehalose 661015840-dimycolate (cord factor) ofMycobacterium tuberculosis inducesforeign-body- and hypersensitivity-type granulomas in micerdquoInfection and Immunity vol 69 no 2 pp 810ndash815 2001

[41] T VGuidry R LHunter Jr and J K Actor ldquoMycobacterial gly-colipid trehalose 661015840-dimycolate-induced hypersensitive gran-ulomas contribution of CD4+ lymphocytesrdquoMicrobiology vol153 no 10 pp 3360ndash3369 2007

[42] E Rhoades F-F Hsu J B Torrelles J Turk D Chatterjee andD G Russell ldquoIdentification and macrophage-activating activ-ity of glycolipids released from intracellular Mycobacteriumbovis BCGrdquoMolecular Microbiology vol 48 no 4 pp 875ndash8882003

8 Mediators of Inflammation

[43] MMHossain andM-N Norazmi ldquoPattern recognition recep-tors and cytokines in Mycobacterium tuberculosis infectionmdashthe double-edged swordrdquo BioMed Research International vol2013 Article ID 179174 18 pages 2013

[44] D M E Bowdish K Sakamoto M-J Kim et al ldquoMARCOTLR2 and CD14 are required for macrophage cytokine res-ponses to mycobacterial trehalose dimycolate and Mycobac-terium tuberculosisrdquo PLoS Pathogens vol 5 no 6 Article IDe1000474 2009

[45] E Mortaz I M Adcock P Tabarsi et al ldquoInteraction of patternrecognition receptors withMycobacterium tuberculosisrdquo Journalof Clinical Immunology vol 35 no 1 pp 1ndash10 2015

[46] D G Russell ldquoWho puts the tubercle in tuberculosisrdquo NatureReviews Microbiology vol 5 no 1 pp 39ndash47 2007

[47] H Schoenen B Bodendorfer K Hitchens et al ldquoCutting Edgemincle is essential for recognition and adjuvanticity of themycobacterial cord factor and its synthetic analog trehalose-dibehenaterdquo Journal of Immunology vol 184 no 6 pp 2756ndash2760 2010

[48] E Ishikawa T Ishikawa Y S Morita et al ldquoDirect recognitionof themycobacterial glycolipid trehalose dimycolate by C-typelectinMinclerdquo Journal of ExperimentalMedicine vol 206 no 13pp 2879ndash2888 2009

[49] I Matsunaga and D B Moody ldquoMincle is a long soughtreceptor for mycobacterial cord factorrdquo Journal of ExperimentalMedicine vol 206 no 13 pp 2865ndash2868 2009

[50] K Werninghaus A Babiak O Gross et al ldquoAdjuvanticity ofa synthetic cord factor analogue for subunit Mycobacteriumtuberculosis vaccination requires FclR120574-Syk-Card9-dependentinnate immune activationrdquo Journal of Experimental Medicinevol 206 no 1 pp 89ndash97 2009

[51] S LeibundGut-LandmannOGrossM J Robinson et al ldquoSyk-and CARD9-dependent coupling of innate immunity to theinduction of T helper cells that produce interleukin 17rdquo NatureImmunology vol 8 no 6 pp 630ndash638 2007

[52] J Indrigo R L Hunter Jr and J K Actor ldquoInfluence of tre-halose 661015840-dimycolate (TDM) during mycobacterial infectionof bone marrow macrophagesrdquoMicrobiology vol 148 no 7 pp1991ndash1998 2002

[53] J Indrigo R L Hunter Jr and J K Actor ldquoCord factortrehalose 661015840-dimycolate (TDM) mediates trafficking eventsduringmycobacterial infection ofmurinemacrophagesrdquoMicro-biology vol 149 no 8 pp 2049ndash2059 2003

[54] J K Actor M Olsen R L Hunter Jr and Y-J Geng ldquoDys-regulated response to mycobacterial cord factor trehalose-661015840-dimycolate inCD1Dminusminusmicerdquo Journal of Interferon andCytokineResearch vol 21 no 12 pp 1089ndash1096 2001

[55] J K Actor J Indrigo C M Beachdel et al ldquoMycobacterial gly-colipid cord factor trehalose 661015840-dimycolate causes a decreasein serum cortisol during the granulomatous responserdquo Neuro-ImmunoModulation vol 10 no 5 pp 270ndash282 2002

[56] A N Abbott K J Welsh S-A Hwang et al ldquoIL-6 mediates11120573hsd type 2 to effect progression of the mycobacterial cordfactor trehalose 661015840-dimycolate-induced granulomatous res-ponserdquo NeuroImmunoModulation vol 18 no 4 pp 212ndash2252011

[57] T V Guidry M Olsen K-S Kil R L Hunter Jr Y-J Gengand J K Actor ldquoFailure of CD1Dminusminusmice to elicit hypersensitivegranulomas to mycobacterial cord factor trehalose 661015840-dimy-colaterdquo Journal of Interferon amp Cytokine Research vol 24 no 6pp 362ndash371 2004

[58] TVGuidry R LHunter Jr and J KActor ldquoCD3+ cells transferthe hypersensitive granulomatous response to mycobacterialglycolipid trehalose 661015840-dimycolate in micerdquoMicrobiology vol152 no 12 pp 3765ndash3775 2006

[59] B Lonnerdal and S Iyer ldquoLactoferrin molecular structure andbiological functionrdquoAnnual Review of Nutrition vol 15 pp 93ndash110 1995

[60] L Sanchez M Calvo and J H Brock ldquoBiological role of lacto-ferrinrdquo Archives of Disease in Childhood vol 67 no 5 pp 657ndash661 1992

[61] P Manzoni M Rinaldi S Cattani et al ldquoBovine lactoferrinsupplementation for prevention of late-onset sepsis in verylow-birth-weight neonates a randomized trialrdquo Journal of theAmerican Medical Association vol 302 no 13 pp 1421ndash14282009

[62] S Baveye E Elass J Mazurier G Spik and D Legrand ldquoLacto-ferrin a multifunctional glycoprotein involved in the modula-tion of the inflammatory processrdquoClinical Chemistry and Labo-ratory Medicine vol 37 no 3 pp 281ndash286 1999

[63] R D Baynes and W R Bezwoda ldquoLactoferrin and the inflam-matory responserdquo Advances in Experimental Medicine andBiology vol 357 pp 133ndash141 1994

[64] M Kocieba M Zimecki M Kruzel and J Actor ldquoThe adjuvantactivity of lactoferrin in the generation of DTH to ovalbumincan be inhibited by bovine serum albumin bearing alpha-D-mannopyranosyl residuesrdquoCellularampMolecular Biology Lettersvol 7 no 4 pp 1131ndash1136 2002

[65] C G Turin A Zea-Vera A Pezo et al ldquoLactoferrin for pre-vention of neonatal sepsisrdquo Biometals vol 27 no 5 pp 1007ndash1016 2014

[66] T Siqueiros-Cendon S Arevalo-Gallegos B F Iglesias-Figueroa I A Garcıa-Montoya J Salazar-Martınez and QRascon-Cruz ldquoImmunomodulatory effects of lactoferrinrdquo ActaPharmacologica Sinica vol 35 no 5 pp 557ndash566 2014

[67] D Latorre F Berlutti P Valenti S Gessani and P Puddu ldquoLFimmunomodulatory strategies mastering bacterial endotoxinrdquoBiochemistry and Cell Biology vol 90 no 3 pp 269ndash278 2012

[68] D Legrand ldquoLactoferrin a key molecule in immune andinflammatory processesrdquo Biochemistry and Cell Biology vol 90no 3 pp 252ndash268 2012

[69] S-A Hwang K M Wilk Y A Bangale M L Kruzel and JK Actor ldquoLactoferrin modulation of IL-12 and IL-10 responsefrom activated murine leukocytesrdquo Medical Microbiology andImmunology vol 196 no 3 pp 171ndash180 2007

[70] M Zimecki J Mazurier G Spik and J A Kapp ldquoHuman lacto-ferrin induces phenotypic and functional changes in murinesplenic B cellsrdquo Immunology vol 86 no 1 pp 122ndash127 1995

[71] M Zimecki J Artym G Chodaczek et al ldquoImmunoregulatoryfunction of lactoferrin in immunosuppressed and autoimmuneanimalsrdquo Postepy Higieny i Medycyny Doswiadczalnej vol 61pp 283ndash287 2007

[72] M Zimecki and M L Kruzel ldquoMilk-derived proteins andpeptides of potential therapeutic and nutritive valuerdquo Journal ofExperimental Therapeutics and Oncology vol 6 no 2 pp 89ndash106 2007

[73] M Zimecki RMiędzybrodzki J Mazurier andG Spik ldquoRegu-latory effects of lactoferrin and lipopolysaccharide on LFA-1 expression on human peripheral blood mononuclear cellsrdquoArchivum Immunologiae et Therapiae Experimentalis vol 47no 4 pp 257ndash264 1999

Mediators of Inflammation 9

[74] M L Kruzel and M Zimecki ldquoLactoferrin and immunologicdissonance clinical implicationsrdquo Archivum Immunologiae etTherapiae Experimentalis vol 50 no 6 pp 399ndash410 2002

[75] W I Weis M E Taylor and K Drickamer ldquoThe C-type lectinsuperfamily in the immune systemrdquo Immunological Reviewsvol 163 pp 19ndash34 1998

[76] W I Weis K Drickamer and W A Hendrickson ldquoStructureof a C-type mannose-binding protein complexed with an oli-gosacchariderdquo Nature vol 360 no 6400 pp 127ndash134 1992

[77] T Zagulski Z Jarzabek A Zagulska M Jaszczak I EKochanowska and M Zimecki ldquoLactoferrin stimulates killingand clearance of bacteria but does not prevent mortality of dia-betic micerdquo Archivum Immunologiae et Therapiae Experimen-talis vol 49 no 6 pp 431ndash438 2001

[78] M Kruzel Y Harai C Chen and A Castro ldquoRole of lactoferrinin development of systemic inflammatory response syndrome(SIRS)rdquo in Proceedings of the 2nd International ConferenceProgress in Intensive Care Medicine pp 11ndash12 Wroclaw Poland1998

[79] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoThe guta key metabolic organ protected by lactoferrin during experi-mental systemic inflammation in micerdquo in Advances in Lacto-ferrin Research vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Springer New York NY USA 1998

[80] MKruzel T Zagulski andMZimecki ldquoLactoferrin and insult-induced metabolic imbalance in humans and other animalsrdquoin Lactoferrin Structure Function and Applications Proceedingsof the 4th International Conference on Lactoferrin StructureFunction and Applications held in Sapporo Japan 18ndash22 May1999 pp 301ndash311 ICS Elsevier New York NY USA 2000

[81] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoLactofer-rin protects gut mucosal integrity during endotoxemia inducedby lipopolysaccharide in micerdquo Inflammation vol 24 no 1 pp33ndash44 2000

[82] E Zaczynska M Kocięba E Sliwinska and M ZimeckildquoBovine lactoferrin enhances proliferation of human peripheralblood lymphocytes and induces cytokine production in wholeblood culturesrdquo Advances in Clinical and Experimental Medi-cine vol 23 pp 871ndash876 2014

[83] Z Wei T Nishimura and S Yoshida ldquoCharacterization ofglycans in a lactoferrin isoform lactoferrin-ardquo Journal of DairyScience vol 84 no 12 pp 2584ndash2590 2001

[84] B-K Choi J K Actor S Rios et al ldquoRecombinant humanlactoferrin expressed in glycoengineered Pichia pastoris effectof terminal N-acetylneuraminic acid on in vitro secondaryhumoral immune responserdquoGlycoconjugate Journal vol 25 no6 pp 581ndash593 2008

[85] B Samyn-Petit J-PWajdaDubos F Chirat et al ldquoComparativeanalysis of the site-specific N-glycosylation of human lactofer-rin produced in maize and tobacco plantsrdquo European Journal ofBiochemistry vol 270 no 15 pp 3235ndash3242 2003

[86] M Zimecki J Artym M Kocięba M Duk and M L KruzelldquoThe effect of carbohydrate moiety structure on the immu-noregulatory activity of lactoferrin in vitrordquo Cellular ampMolecu-lar Biology Letters vol 19 no 2 pp 284ndash296 2014

[87] J K Actor S-A Hwang and M L Kruzel ldquoLactoferrin as anatural immune modulatorrdquo Current Pharmaceutical Designvol 15 no 17 pp 1956ndash1973 2009

[88] M L Kruzel J K Actor I Boldogh and M ZimeckildquoLactoferrin in health and diseaserdquo Postepy Higieny i MedycynyDoswiadczalnej vol 61 pp 261ndash267 2007

[89] S Baveye E Elass D G Fernig C Blanquart J Mazurier andD Legrand ldquoHuman lactoferrin interacts with soluble CD14and inhibits expression of endothelial adhesion molecules E-selectin and ICAM-1 induced by the CD14-lipopolysaccharidecomplexrdquo Infection and Immunity vol 68 no 12 pp 6519ndash65252000

[90] C S Curran K P Demick and J M Mansfield ldquoLactoferrinactivates macrophages via TLR4-dependent and -independentsignaling pathwaysrdquo Cellular Immunology vol 242 no 1 pp23ndash30 2006

[91] P Puddu D Latorre M Carollo et al ldquoBovine lactoferrincounteracts Toll-Like receptor mediated activation signals inantigen presenting cellsrdquo PLoS ONE vol 6 no 7 Article IDe22504 2011

[92] T M Doherty and M Arditi ldquoTB or not TB That is the ques-tionmdashdoes TLR signaling hold the answerrdquoThe Journal of Clin-ical Investigation vol 114 no 12 pp 1699ndash1703 2004

[93] M A Rahman P Sobia N Gupta L Van Kaer and GDas ldquoMycobacterium tuberculosis subverts the TLR-2mdashMyD88pathway to facilitate its translocation into the cytosolrdquo PLoSONE vol 9 no 1 Article ID e86886 2014

[94] J ArtymM Zimecki andM L Kruzel ldquoEnhanced clearance ofEscherichia coli and Staphylococcus aureus in mice treated withcyclophosphamide and lactoferrinrdquo International Immunophar-macology vol 4 no 9 pp 1149ndash1157 2004

[95] G Chodaczek A Saavedra-Molina A Bacsi M L Kruzel SSur and I Boldogh ldquoIron-mediated dismutation of superoxideanion augments antigen-induced allergic inflammation effectof lactoferrinrdquo Postepy Higieny i Medycyny Doswiadczalnej vol61 pp 268ndash276 2007

[96] M Kruzel Y Harai C Chen and A Castro ldquoThe Gut a keymetabolic organ protected by lactoferrin during experimentalsystemic inflammation in micerdquo in Advances in LactoferrinResearch G Spik D Legrand J Mazurier A Pierce and J-PPerraudin Eds vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Plenum Press 1998

[97] M L Kruzel A Bacsi B Choudhury S Sur and I BoldoghldquoLactoferrin decreases pollen antigen-induced allergic airwayinflammation in a murine model of asthmardquo Immunology vol119 no 2 pp 159ndash166 2006

[98] J K Actor S-A Hwang M Olsen M Zimecki R L HunterJr and M L Kruzel ldquoLactoferrin immunomodulation of DTHresponse in micerdquo International Immunopharmacology vol 2no 4 pp 475ndash486 2002

[99] S-A Hwang M L Kruzel and J K Actor ldquoInfluence of bovinelactoferrin on expression of presentation molecules on BCG-infected bonemarrow derivedmacrophagesrdquo Biochimie vol 91no 1 pp 76ndash85 2009

[100] M Zimecki J Mazurier M Machnicki Z Wieczorek JMontreuil and G Spik ldquoImmunostimulatory activity of lacto-transferrin andmaturation of CD4minus CD8minusmurine thymocytesrdquoImmunology Letters vol 30 no 1 pp 119ndash123 1991

[101] K J Welsh S-A Hwang R L Hunter M L Kruzel and JK Actor ldquoLactoferrin modulation of mycobacterial cord factortrehalose 6-6rsquo-dimycolate induced granulomatous responserdquoTranslational Research vol 156 no 4 pp 207ndash215 2010

[102] M L Bourigault N Segueni S Rose et al ldquoRelative contri-bution of IL-1alpha IL-1beta and TNF to the host responseto Mycobacterium tuberculosis and attenuated M bovis BCGrdquoImmunity Inflammation and Disease vol 1 no 1 pp 47ndash622013

10 Mediators of Inflammation

[103] C Holscher R A Atkinson B Arendse et al ldquoA protective andagonistic function of IL-12p40 in mycobacterial infectionrdquoTheJournal of Immunology vol 167 no 12 pp 6957ndash6966 2001

[104] M Jacobs L Fick N Allie N Brown and B Ryffel ldquoEnhancedimmune response in Mycobacterium bovis bacille calmetteguerin (BCG)-infected IL-10-deficientmicerdquoClinical Chemistryand Laboratory Medicine vol 40 no 9 pp 893ndash902 2002

[105] K J Welsh S-A Hwang S Boyd M L Kruzel R L Hunterand J K Actor ldquoInfluence of oral lactoferrin onMycobacteriumtuberculosis induced immunopathologyrdquo Tuberculosis vol 91supplement 1 pp S105ndashS113 2011

[106] M L Kruzel J K Actor M Zimecki et al ldquoNovel recombinanthuman lactoferrin differential activation of oxidative stressrelated gene expressionrdquo Journal of Biotechnology vol 168 no4 pp 666ndash675 2013

[107] Y Shibata J Gabbard M Yamashita et al ldquoHeat-killedBCG induces biphasic cyclooxygenase 2+ splenic macrophageformationmdashrole of IL-10 and bone marrow precursorsrdquo Journalof Leukocyte Biology vol 80 no 3 pp 590ndash598 2006

[108] H Yamada E Kuroda S Matsumoto T Matsumoto TYamada and U Yamashita ldquoProstaglandin E

2down-regulates

viable Bacille Calmette-Guerin-induced macrophage cytotoxi-city against murine bladder cancer cell MBT-2 in vitrordquoClinicaland Experimental Immunology vol 128 no 1 pp 52ndash58 2002

[109] S Hwang M L Kruzel and J K Actor ldquoEffects of CHO-expressed recombinant lactoferrins on mouse dendritic cellpresentation and functionrdquo Innate Immunity vol 21 no 5 pp553ndash561 2015

[110] K M OShea S-A Hwang and J K Actor ldquoImmune activity ofbcg infected mouse macrophages treated with a novel recom-binant mouse lactoferrinrdquo Annals of Clinical Lab Science vol45 no 5 pp 403ndash410 2015

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Oxidative Medicine and Cellular Longevity

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PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Research and TreatmentAIDS

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Mediators of Inflammation 5

2000

1500

1000

500

0

TDM

Treatment

lowast lowast lowast lowast

IL-12

p40

conc

entr

atio

n (p

glu

ng)

Lung IL-12p40

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

6000

2000

3000

4000

5000

1000

0

TDM

Treatment

lowast lowast lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

Lung TNF-120572

IL-6

conc

entr

atio

n (p

glu

ng)

2000

1500

1000

500

0

TDM

Treatment

lowast lowast lowast

Lung IL-6

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

40000

20000

30000

10000

0

TDM

Treatment

lowast lowast lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

IL-1120573

conc

entr

atio

n (p

glu

ng)

Lung IL-1120573

IL-10

conc

entr

atio

n (p

glu

ng)

Lung IL-10300

200

100

0

TDM

Treatment

lowast

Naiumlve TDM

1mgbovine

TDM

2mgbovine

TDM

1mgrHuLF

TDM

2mgrHuLF

TNF-120572

conc

entr

atio

n (p

glu

ng)

Figure 3 Proinflammatory cytokines in lungs are reduced in lactoferrin-treated mice after challenge with TDM BALBc mice werechallenged with 25120583g TDM in a water-in-oil emulsion Bovine or recombinant human lactoferrin (1 or 2mg) was orally administered at4 and 6 days after TDM challenge Methods of TDM induction are described elsewhere [38 101] Lungs were isolated on day 7 after TDMchallenge cytokines were assessed by ELISA values are shown per lung for individual mice lowast119901 lt 005 compared to the TDM emulsion alonetreated mice

of antimycobacterial drugs to sites harboring infectiousagents in essence permitting an efficient mechanism totreat TB infection Overall these studies permit confidenceto target clinical implications and test novel strategies to

modulate MTB-induced granulomatous responses to allowmore efficacious delivery of therapeutics to mycobacterialorganisms residing within lung tissue during activemycobac-terial disease states

6 Mediators of Inflammation

BALB

cC5

7BL

6TDM alone TDM + LFNaiumlve

TDM alone TDM + LFNaiumlve

Figure 4 Granuloma response to TDM challenge in oral recombinant human lactoferrin-treated mice Two strains of mice (BALBc andC57BL6) were challenged with 25120583g TDM in a water-in-oil emulsion as described [38 101] Human lactoferrin (recombinant 1mg) wasgiven orally at 4 and 6 days after TDM challenge Comparisons are made to naıve mice and to mice only administered TDM HampE staining100x Representative sections from mice at 7 days after TDM challenge119873 ge 8 per experimental group

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

This work was supported in part by NIH Grants 1R41-AI117990-01 and R42-AI051050-05The authors acknowledgePharmaReview Corp (Houston TX) for their kind gifts oflactoferrins and thank Marian L Kruzel PhD and Shen-An Hwang PhD for their insights into the research projectThe authors also thank Ron Gadot and Ai Yamazaki for theirtechnical contributions to the research presented

References

[1] P E M Fine ldquoHerd immunity history theory practicerdquo Epide-miologic Reviews vol 15 no 2 pp 265ndash302 1993

[2] L Ramakrishnan ldquoRevisiting the role of the granuloma intuberculosisrdquo Nature Reviews Immunology vol 12 no 5 pp352ndash366 2012

[3] M Silva Miranda A Breiman S Allain F Deknuydt andF Altare ldquoThe tuberculous granuloma an unsuccessful hostdefence mechanism providing a safety shelter for the bacteriardquoClinical amp Developmental Immunology vol 2012 Article ID139127 14 pages 2012

[4] S A Khader J Rangel-Moreno J J Fountain et al ldquoIn amurinetuberculosis model the absence of homeostatic chemokinesdelays granuloma formation and protective immunityrdquo Journalof Immunology vol 183 no 12 pp 8004ndash8014 2009

[5] AMCooper and S A Khader ldquoThe role of cytokines in the ini-tiation expansion and control of cellular immunity to tubercu-losisrdquo Immunological Reviews vol 226 no 1 pp 191ndash204 2008

[6] IMOrme and R J Basaraba ldquoThe formation of the granulomain tuberculosis infectionrdquo Seminars in Immunology vol 26 no6 pp 601ndash609 2014

[7] I M Orme R T Robinson and A M Cooper ldquoThe balancebetween protective and pathogenic immune responses in theTB-infected lungrdquo Nature Immunology vol 16 no 1 pp 57ndash632014

[8] J K Actor M Olsen C Jagannath and R L Hunter ldquoRela-tionship of survival organism containment and granulomaformation in acute murine tuberculosisrdquo Journal of Interferonand Cytokine Research vol 19 no 10 pp 1183ndash1193 1999

[9] J L Flynn and J Chan ldquoWhatrsquos good for the host is good for thebugrdquo Trends in Microbiology vol 13 no 3 pp 98ndash102 2005

[10] D G Russell ldquoMycobacterium tuberculosis and the intimatediscourse of a chronic infectionrdquo Immunological Reviews vol240 no 1 pp 252ndash268 2011

[11] R L Hunter ldquoPathology of post primary tuberculosis of thelung an illustrated critical reviewrdquo Tuberculosis vol 91 no 6pp 497ndash509 2011

[12] R L Hunter J K Actor S-A Hwang V Karev and C Jagan-nath ldquoPathogenesis of post primary tuberculosis immunity andhypersensitivity in the development of cavitiesrdquoAnnals of Clini-cal and Laboratory Science vol 44 no 4 pp 365ndash387 2014

[13] K J Welsh R L Hunter and J K Actor ldquoTrehalose 661015840-dimy-colatemdasha coat to regulate tuberculosis immunopathogenesisrdquoTuberculosis vol 93 supplement pp S3ndashS9 2013

[14] M-J Kim H C Wainwright M Locketz et al ldquoCaseation ofhuman tuberculosis granulomas correlates with elevated hostlipid metabolismrdquo EMBO Molecular Medicine vol 2 no 7 pp258ndash274 2010

[15] J K Actor E Breij R A Wetsel H Hoffmann R L HunterJr and C Jagannath ldquoA role for complement C5 in organ-ism containment and granulomatous response during murine

Mediators of Inflammation 7

tuberculosisrdquo Scandinavian Journal of Immunology vol 53 no5 pp 464ndash474 2001

[16] J K Actor C D Leonard V E Watson et al ldquoCytokinemRNAexpression and serumcortisol evaluation duringmurinelung inflammation induced by Mycobacterium tuberculosisrdquoCombinatorial Chemistry and High Throughput Screening vol3 no 4 pp 343ndash351 2000

[17] C W Borders A Courtney K Ronen et al ldquoRequisite role forcomplement C5 and the C5a receptor in granulomatous res-ponse to mycobacterial glycolipid trehalose 661015840-dimycolaterdquoScandinavian Journal of Immunology vol 62 no 2 pp 123ndash1302005

[18] R L Hunter C Jagannath and J K Actor ldquoPathology of post-primary tuberculosis in humans and mice contradiction oflong-held beliefsrdquo Tuberculosis vol 87 no 4 pp 267ndash278 2007

[19] C Jagannath H Hoffmann E Sepulveda J K Actor R AWetsel and R L Hunter Jr ldquoHypersusceptibility of AJ miceto tuberculosis is in part due to a deficiency of the fifth comple-ment component (C5)rdquo Scandinavian Journal of Immunologyvol 52 no 4 pp 369ndash379 2000

[20] V E Watson L L Hill L B Owen-Schaub et al ldquoApoptosis inMycobacterium tuberculosis infection in mice exhibiting variedimmunopathologyrdquo Journal of Pathology vol 190 no 2 pp 211ndash220 2000

[21] R H Copenhaver E Sepulveda L Y Armitige et al ldquoAmutantof Mycobacterium tuberculosis H37Rv that lacks expressionof antigen 85A is attenuated in mice but retains vaccinogenicpotentialrdquo Infection and Immunity vol 72 no 12 pp 7084ndash7095 2004

[22] S-A Hwang R Arora M L Kruzel and J K Actor ldquoLactofer-rin enhances efficacy of the BCG vaccine comparison betweentwo inbred mice strains (C57BL6 and BALBc)rdquo Tuberculosisvol 89 supplement 1 pp S49ndashS54 2009

[23] S-A Hwang K Wilk M L Kruzel and J K Actor ldquoA novelrecombinant human lactoferrin augments the BCG vaccine andprotects alveolar integrity upon infection with Mycobacteriumtuberculosis in micerdquo Vaccine vol 27 no 23 pp 3026ndash30342009

[24] S-A Hwang K M Wilk M Budnicka et al ldquoLactoferrinenhanced efficacy of the BCG vaccine to generate host protec-tive responses against challenge with virulent Mycobacteriumtuberculosisrdquo Vaccine vol 25 no 37-38 pp 6730ndash6743 2007

[25] S-A Hwang and J K Actor ldquoLactoferrin modulation of BCG-infected dendritic cell functionsrdquo International Immunologyvol 21 no 10 pp 1185ndash1197 2009

[26] S-A Hwang M L Kruzel and J K Actor ldquoLactoferrin aug-ments BCG vaccine efficacy to generate T helper response andsubsequent protection against challenge with virulentMycobac-terium tuberculosisrdquo International Immunopharmacology vol 5no 3 pp 591ndash599 2005

[27] K M Wilk S-A Hwang and J K Actor ldquoLactoferrin mod-ulation of antigen-presenting-cell response to BCG infectionrdquoPostepy Higieny i Medycyny Doswiadczalnej vol 61 pp 277ndash282 2007

[28] S A Hwang M L Kruzel and J K Actor ldquoCHO expressedrecombinant human lactoferrin as an adjuvant for BCGrdquoInternational Journal of Immunopathology and Pharmacologyvol 28 no 4 pp 452ndash468 2015

[29] R L Perez J Roman G W Staton Jr and R L HunterldquoExtravascular coagulation and fibrinolysis in murine lunginflammation induced by the mycobacterial cord factor

trehalose-66rsquo-dimycolaterdquoAmerican Journal of Respiratory andCritical Care Medicine vol 149 no 2 pp 510ndash518 1994

[30] C A Behling R L Perez M R Kidd G W Staton Jr and RL Hunter ldquoInduction of pulmonary granulomas macrophageprocoagulant activity and tumor necrosis factor-alpha by tre-halose glycolipidsrdquo Annals of Clinical and Laboratory Sciencevol 23 no 4 pp 256ndash266 1993

[31] G S Retzinger S C Meredith R L Hunter K Takayama andF J Kezdy ldquoIdentification of the physiologically active state ofthe mycobacterial glycolipid trehalose 661015840-dimycolate and therole of fibrinogen in the biologic activities of trehalose 661015840-dimycolate monolayersrdquo Journal of Immunology vol 129 no 2pp 735ndash744 1982

[32] G S Retzinger S C Meredith K Takayama R L Hunter andF J Kezdy ldquoThe role of surface in the biological activities oftrehalose 661015840-dimycolate Surface properties and developmentof amodel systemrdquoThe Journal of Biological Chemistry vol 256no 15 pp 8208ndash8216 1981

[33] R L Perez J Roman S Roser et al ldquoCytokinemessage and pro-tein expression during lung granuloma formation and resolu-tion induced by the mycobacterial cord factor trehalose-661015840-dimycolaterdquo Journal of Interferon amp Cytokine Research vol 20no 9 pp 795ndash804 2000

[34] R L Hunter M R Olsen C Jagannath and J K Actor ldquoMult-iple roles of cord factor in the pathogenesis of primary sec-ondary and cavitary tuberculosis including a revised descrip-tion of the pathology of secondary diseaserdquo Annals of Clinicaland Laboratory Science vol 36 no 4 pp 371ndash386 2006

[35] J K Actor ldquoMycobacterial TDM a coat to modulate post pri-mary pathogenesisrdquoMycobacterial Diseases vol 2 no 1 articlee106 2012

[36] R L HunterMOlsen C Jagannath and J K Actor ldquoTrehalose661015840-dimycolate and lipid in the pathogenesis of caseatinggranulomas of tuberculosis in micerdquo The American Journal ofPathology vol 168 no 4 pp 1249ndash1261 2006

[37] A N Abbott T V Guidry K J Welsh et al ldquo11Beta-hydroxy-steroid dehydrogenases are regulated during the pulmonarygranulomatous response to the mycobacterial glycolipidtrehalose-661015840-dimycolaterdquo NeuroImmunoModulation vol 16no 3 pp 147ndash154 2009

[38] K J Welsh A N Abbott S-A Hwang et al ldquoA role for tumournecrosis factor-120572 complement C5 and interleukin-6 in theinitiation and development of the mycobacterial cord factortrehalose 661015840-dimycolate induced granulomatous responserdquoMicrobiology vol 154 no 6 pp 1813ndash1824 2008

[39] A Bekierkunst and E Yarkoni ldquoGranulomatous hypersensitiv-ity to trehalose-66rsquo-dimycolate (cord factor) in mice infectedwith BCGrdquo Infection and Immunity vol 7 no 4 pp 631ndash6381973

[40] H Yamagami TMatsumoto N Fujiwara et al ldquoTrehalose 661015840-dimycolate (cord factor) ofMycobacterium tuberculosis inducesforeign-body- and hypersensitivity-type granulomas in micerdquoInfection and Immunity vol 69 no 2 pp 810ndash815 2001

[41] T VGuidry R LHunter Jr and J K Actor ldquoMycobacterial gly-colipid trehalose 661015840-dimycolate-induced hypersensitive gran-ulomas contribution of CD4+ lymphocytesrdquoMicrobiology vol153 no 10 pp 3360ndash3369 2007

[42] E Rhoades F-F Hsu J B Torrelles J Turk D Chatterjee andD G Russell ldquoIdentification and macrophage-activating activ-ity of glycolipids released from intracellular Mycobacteriumbovis BCGrdquoMolecular Microbiology vol 48 no 4 pp 875ndash8882003

8 Mediators of Inflammation

[43] MMHossain andM-N Norazmi ldquoPattern recognition recep-tors and cytokines in Mycobacterium tuberculosis infectionmdashthe double-edged swordrdquo BioMed Research International vol2013 Article ID 179174 18 pages 2013

[44] D M E Bowdish K Sakamoto M-J Kim et al ldquoMARCOTLR2 and CD14 are required for macrophage cytokine res-ponses to mycobacterial trehalose dimycolate and Mycobac-terium tuberculosisrdquo PLoS Pathogens vol 5 no 6 Article IDe1000474 2009

[45] E Mortaz I M Adcock P Tabarsi et al ldquoInteraction of patternrecognition receptors withMycobacterium tuberculosisrdquo Journalof Clinical Immunology vol 35 no 1 pp 1ndash10 2015

[46] D G Russell ldquoWho puts the tubercle in tuberculosisrdquo NatureReviews Microbiology vol 5 no 1 pp 39ndash47 2007

[47] H Schoenen B Bodendorfer K Hitchens et al ldquoCutting Edgemincle is essential for recognition and adjuvanticity of themycobacterial cord factor and its synthetic analog trehalose-dibehenaterdquo Journal of Immunology vol 184 no 6 pp 2756ndash2760 2010

[48] E Ishikawa T Ishikawa Y S Morita et al ldquoDirect recognitionof themycobacterial glycolipid trehalose dimycolate by C-typelectinMinclerdquo Journal of ExperimentalMedicine vol 206 no 13pp 2879ndash2888 2009

[49] I Matsunaga and D B Moody ldquoMincle is a long soughtreceptor for mycobacterial cord factorrdquo Journal of ExperimentalMedicine vol 206 no 13 pp 2865ndash2868 2009

[50] K Werninghaus A Babiak O Gross et al ldquoAdjuvanticity ofa synthetic cord factor analogue for subunit Mycobacteriumtuberculosis vaccination requires FclR120574-Syk-Card9-dependentinnate immune activationrdquo Journal of Experimental Medicinevol 206 no 1 pp 89ndash97 2009

[51] S LeibundGut-LandmannOGrossM J Robinson et al ldquoSyk-and CARD9-dependent coupling of innate immunity to theinduction of T helper cells that produce interleukin 17rdquo NatureImmunology vol 8 no 6 pp 630ndash638 2007

[52] J Indrigo R L Hunter Jr and J K Actor ldquoInfluence of tre-halose 661015840-dimycolate (TDM) during mycobacterial infectionof bone marrow macrophagesrdquoMicrobiology vol 148 no 7 pp1991ndash1998 2002

[53] J Indrigo R L Hunter Jr and J K Actor ldquoCord factortrehalose 661015840-dimycolate (TDM) mediates trafficking eventsduringmycobacterial infection ofmurinemacrophagesrdquoMicro-biology vol 149 no 8 pp 2049ndash2059 2003

[54] J K Actor M Olsen R L Hunter Jr and Y-J Geng ldquoDys-regulated response to mycobacterial cord factor trehalose-661015840-dimycolate inCD1Dminusminusmicerdquo Journal of Interferon andCytokineResearch vol 21 no 12 pp 1089ndash1096 2001

[55] J K Actor J Indrigo C M Beachdel et al ldquoMycobacterial gly-colipid cord factor trehalose 661015840-dimycolate causes a decreasein serum cortisol during the granulomatous responserdquo Neuro-ImmunoModulation vol 10 no 5 pp 270ndash282 2002

[56] A N Abbott K J Welsh S-A Hwang et al ldquoIL-6 mediates11120573hsd type 2 to effect progression of the mycobacterial cordfactor trehalose 661015840-dimycolate-induced granulomatous res-ponserdquo NeuroImmunoModulation vol 18 no 4 pp 212ndash2252011

[57] T V Guidry M Olsen K-S Kil R L Hunter Jr Y-J Gengand J K Actor ldquoFailure of CD1Dminusminusmice to elicit hypersensitivegranulomas to mycobacterial cord factor trehalose 661015840-dimy-colaterdquo Journal of Interferon amp Cytokine Research vol 24 no 6pp 362ndash371 2004

[58] TVGuidry R LHunter Jr and J KActor ldquoCD3+ cells transferthe hypersensitive granulomatous response to mycobacterialglycolipid trehalose 661015840-dimycolate in micerdquoMicrobiology vol152 no 12 pp 3765ndash3775 2006

[59] B Lonnerdal and S Iyer ldquoLactoferrin molecular structure andbiological functionrdquoAnnual Review of Nutrition vol 15 pp 93ndash110 1995

[60] L Sanchez M Calvo and J H Brock ldquoBiological role of lacto-ferrinrdquo Archives of Disease in Childhood vol 67 no 5 pp 657ndash661 1992

[61] P Manzoni M Rinaldi S Cattani et al ldquoBovine lactoferrinsupplementation for prevention of late-onset sepsis in verylow-birth-weight neonates a randomized trialrdquo Journal of theAmerican Medical Association vol 302 no 13 pp 1421ndash14282009

[62] S Baveye E Elass J Mazurier G Spik and D Legrand ldquoLacto-ferrin a multifunctional glycoprotein involved in the modula-tion of the inflammatory processrdquoClinical Chemistry and Labo-ratory Medicine vol 37 no 3 pp 281ndash286 1999

[63] R D Baynes and W R Bezwoda ldquoLactoferrin and the inflam-matory responserdquo Advances in Experimental Medicine andBiology vol 357 pp 133ndash141 1994

[64] M Kocieba M Zimecki M Kruzel and J Actor ldquoThe adjuvantactivity of lactoferrin in the generation of DTH to ovalbumincan be inhibited by bovine serum albumin bearing alpha-D-mannopyranosyl residuesrdquoCellularampMolecular Biology Lettersvol 7 no 4 pp 1131ndash1136 2002

[65] C G Turin A Zea-Vera A Pezo et al ldquoLactoferrin for pre-vention of neonatal sepsisrdquo Biometals vol 27 no 5 pp 1007ndash1016 2014

[66] T Siqueiros-Cendon S Arevalo-Gallegos B F Iglesias-Figueroa I A Garcıa-Montoya J Salazar-Martınez and QRascon-Cruz ldquoImmunomodulatory effects of lactoferrinrdquo ActaPharmacologica Sinica vol 35 no 5 pp 557ndash566 2014

[67] D Latorre F Berlutti P Valenti S Gessani and P Puddu ldquoLFimmunomodulatory strategies mastering bacterial endotoxinrdquoBiochemistry and Cell Biology vol 90 no 3 pp 269ndash278 2012

[68] D Legrand ldquoLactoferrin a key molecule in immune andinflammatory processesrdquo Biochemistry and Cell Biology vol 90no 3 pp 252ndash268 2012

[69] S-A Hwang K M Wilk Y A Bangale M L Kruzel and JK Actor ldquoLactoferrin modulation of IL-12 and IL-10 responsefrom activated murine leukocytesrdquo Medical Microbiology andImmunology vol 196 no 3 pp 171ndash180 2007

[70] M Zimecki J Mazurier G Spik and J A Kapp ldquoHuman lacto-ferrin induces phenotypic and functional changes in murinesplenic B cellsrdquo Immunology vol 86 no 1 pp 122ndash127 1995

[71] M Zimecki J Artym G Chodaczek et al ldquoImmunoregulatoryfunction of lactoferrin in immunosuppressed and autoimmuneanimalsrdquo Postepy Higieny i Medycyny Doswiadczalnej vol 61pp 283ndash287 2007

[72] M Zimecki and M L Kruzel ldquoMilk-derived proteins andpeptides of potential therapeutic and nutritive valuerdquo Journal ofExperimental Therapeutics and Oncology vol 6 no 2 pp 89ndash106 2007

[73] M Zimecki RMiędzybrodzki J Mazurier andG Spik ldquoRegu-latory effects of lactoferrin and lipopolysaccharide on LFA-1 expression on human peripheral blood mononuclear cellsrdquoArchivum Immunologiae et Therapiae Experimentalis vol 47no 4 pp 257ndash264 1999

Mediators of Inflammation 9

[74] M L Kruzel and M Zimecki ldquoLactoferrin and immunologicdissonance clinical implicationsrdquo Archivum Immunologiae etTherapiae Experimentalis vol 50 no 6 pp 399ndash410 2002

[75] W I Weis M E Taylor and K Drickamer ldquoThe C-type lectinsuperfamily in the immune systemrdquo Immunological Reviewsvol 163 pp 19ndash34 1998

[76] W I Weis K Drickamer and W A Hendrickson ldquoStructureof a C-type mannose-binding protein complexed with an oli-gosacchariderdquo Nature vol 360 no 6400 pp 127ndash134 1992

[77] T Zagulski Z Jarzabek A Zagulska M Jaszczak I EKochanowska and M Zimecki ldquoLactoferrin stimulates killingand clearance of bacteria but does not prevent mortality of dia-betic micerdquo Archivum Immunologiae et Therapiae Experimen-talis vol 49 no 6 pp 431ndash438 2001

[78] M Kruzel Y Harai C Chen and A Castro ldquoRole of lactoferrinin development of systemic inflammatory response syndrome(SIRS)rdquo in Proceedings of the 2nd International ConferenceProgress in Intensive Care Medicine pp 11ndash12 Wroclaw Poland1998

[79] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoThe guta key metabolic organ protected by lactoferrin during experi-mental systemic inflammation in micerdquo in Advances in Lacto-ferrin Research vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Springer New York NY USA 1998

[80] MKruzel T Zagulski andMZimecki ldquoLactoferrin and insult-induced metabolic imbalance in humans and other animalsrdquoin Lactoferrin Structure Function and Applications Proceedingsof the 4th International Conference on Lactoferrin StructureFunction and Applications held in Sapporo Japan 18ndash22 May1999 pp 301ndash311 ICS Elsevier New York NY USA 2000

[81] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoLactofer-rin protects gut mucosal integrity during endotoxemia inducedby lipopolysaccharide in micerdquo Inflammation vol 24 no 1 pp33ndash44 2000

[82] E Zaczynska M Kocięba E Sliwinska and M ZimeckildquoBovine lactoferrin enhances proliferation of human peripheralblood lymphocytes and induces cytokine production in wholeblood culturesrdquo Advances in Clinical and Experimental Medi-cine vol 23 pp 871ndash876 2014

[83] Z Wei T Nishimura and S Yoshida ldquoCharacterization ofglycans in a lactoferrin isoform lactoferrin-ardquo Journal of DairyScience vol 84 no 12 pp 2584ndash2590 2001

[84] B-K Choi J K Actor S Rios et al ldquoRecombinant humanlactoferrin expressed in glycoengineered Pichia pastoris effectof terminal N-acetylneuraminic acid on in vitro secondaryhumoral immune responserdquoGlycoconjugate Journal vol 25 no6 pp 581ndash593 2008

[85] B Samyn-Petit J-PWajdaDubos F Chirat et al ldquoComparativeanalysis of the site-specific N-glycosylation of human lactofer-rin produced in maize and tobacco plantsrdquo European Journal ofBiochemistry vol 270 no 15 pp 3235ndash3242 2003

[86] M Zimecki J Artym M Kocięba M Duk and M L KruzelldquoThe effect of carbohydrate moiety structure on the immu-noregulatory activity of lactoferrin in vitrordquo Cellular ampMolecu-lar Biology Letters vol 19 no 2 pp 284ndash296 2014

[87] J K Actor S-A Hwang and M L Kruzel ldquoLactoferrin as anatural immune modulatorrdquo Current Pharmaceutical Designvol 15 no 17 pp 1956ndash1973 2009

[88] M L Kruzel J K Actor I Boldogh and M ZimeckildquoLactoferrin in health and diseaserdquo Postepy Higieny i MedycynyDoswiadczalnej vol 61 pp 261ndash267 2007

[89] S Baveye E Elass D G Fernig C Blanquart J Mazurier andD Legrand ldquoHuman lactoferrin interacts with soluble CD14and inhibits expression of endothelial adhesion molecules E-selectin and ICAM-1 induced by the CD14-lipopolysaccharidecomplexrdquo Infection and Immunity vol 68 no 12 pp 6519ndash65252000

[90] C S Curran K P Demick and J M Mansfield ldquoLactoferrinactivates macrophages via TLR4-dependent and -independentsignaling pathwaysrdquo Cellular Immunology vol 242 no 1 pp23ndash30 2006

[91] P Puddu D Latorre M Carollo et al ldquoBovine lactoferrincounteracts Toll-Like receptor mediated activation signals inantigen presenting cellsrdquo PLoS ONE vol 6 no 7 Article IDe22504 2011

[92] T M Doherty and M Arditi ldquoTB or not TB That is the ques-tionmdashdoes TLR signaling hold the answerrdquoThe Journal of Clin-ical Investigation vol 114 no 12 pp 1699ndash1703 2004

[93] M A Rahman P Sobia N Gupta L Van Kaer and GDas ldquoMycobacterium tuberculosis subverts the TLR-2mdashMyD88pathway to facilitate its translocation into the cytosolrdquo PLoSONE vol 9 no 1 Article ID e86886 2014

[94] J ArtymM Zimecki andM L Kruzel ldquoEnhanced clearance ofEscherichia coli and Staphylococcus aureus in mice treated withcyclophosphamide and lactoferrinrdquo International Immunophar-macology vol 4 no 9 pp 1149ndash1157 2004

[95] G Chodaczek A Saavedra-Molina A Bacsi M L Kruzel SSur and I Boldogh ldquoIron-mediated dismutation of superoxideanion augments antigen-induced allergic inflammation effectof lactoferrinrdquo Postepy Higieny i Medycyny Doswiadczalnej vol61 pp 268ndash276 2007

[96] M Kruzel Y Harai C Chen and A Castro ldquoThe Gut a keymetabolic organ protected by lactoferrin during experimentalsystemic inflammation in micerdquo in Advances in LactoferrinResearch G Spik D Legrand J Mazurier A Pierce and J-PPerraudin Eds vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Plenum Press 1998

[97] M L Kruzel A Bacsi B Choudhury S Sur and I BoldoghldquoLactoferrin decreases pollen antigen-induced allergic airwayinflammation in a murine model of asthmardquo Immunology vol119 no 2 pp 159ndash166 2006

[98] J K Actor S-A Hwang M Olsen M Zimecki R L HunterJr and M L Kruzel ldquoLactoferrin immunomodulation of DTHresponse in micerdquo International Immunopharmacology vol 2no 4 pp 475ndash486 2002

[99] S-A Hwang M L Kruzel and J K Actor ldquoInfluence of bovinelactoferrin on expression of presentation molecules on BCG-infected bonemarrow derivedmacrophagesrdquo Biochimie vol 91no 1 pp 76ndash85 2009

[100] M Zimecki J Mazurier M Machnicki Z Wieczorek JMontreuil and G Spik ldquoImmunostimulatory activity of lacto-transferrin andmaturation of CD4minus CD8minusmurine thymocytesrdquoImmunology Letters vol 30 no 1 pp 119ndash123 1991

[101] K J Welsh S-A Hwang R L Hunter M L Kruzel and JK Actor ldquoLactoferrin modulation of mycobacterial cord factortrehalose 6-6rsquo-dimycolate induced granulomatous responserdquoTranslational Research vol 156 no 4 pp 207ndash215 2010

[102] M L Bourigault N Segueni S Rose et al ldquoRelative contri-bution of IL-1alpha IL-1beta and TNF to the host responseto Mycobacterium tuberculosis and attenuated M bovis BCGrdquoImmunity Inflammation and Disease vol 1 no 1 pp 47ndash622013

10 Mediators of Inflammation

[103] C Holscher R A Atkinson B Arendse et al ldquoA protective andagonistic function of IL-12p40 in mycobacterial infectionrdquoTheJournal of Immunology vol 167 no 12 pp 6957ndash6966 2001

[104] M Jacobs L Fick N Allie N Brown and B Ryffel ldquoEnhancedimmune response in Mycobacterium bovis bacille calmetteguerin (BCG)-infected IL-10-deficientmicerdquoClinical Chemistryand Laboratory Medicine vol 40 no 9 pp 893ndash902 2002

[105] K J Welsh S-A Hwang S Boyd M L Kruzel R L Hunterand J K Actor ldquoInfluence of oral lactoferrin onMycobacteriumtuberculosis induced immunopathologyrdquo Tuberculosis vol 91supplement 1 pp S105ndashS113 2011

[106] M L Kruzel J K Actor M Zimecki et al ldquoNovel recombinanthuman lactoferrin differential activation of oxidative stressrelated gene expressionrdquo Journal of Biotechnology vol 168 no4 pp 666ndash675 2013

[107] Y Shibata J Gabbard M Yamashita et al ldquoHeat-killedBCG induces biphasic cyclooxygenase 2+ splenic macrophageformationmdashrole of IL-10 and bone marrow precursorsrdquo Journalof Leukocyte Biology vol 80 no 3 pp 590ndash598 2006

[108] H Yamada E Kuroda S Matsumoto T Matsumoto TYamada and U Yamashita ldquoProstaglandin E

2down-regulates

viable Bacille Calmette-Guerin-induced macrophage cytotoxi-city against murine bladder cancer cell MBT-2 in vitrordquoClinicaland Experimental Immunology vol 128 no 1 pp 52ndash58 2002

[109] S Hwang M L Kruzel and J K Actor ldquoEffects of CHO-expressed recombinant lactoferrins on mouse dendritic cellpresentation and functionrdquo Innate Immunity vol 21 no 5 pp553ndash561 2015

[110] K M OShea S-A Hwang and J K Actor ldquoImmune activity ofbcg infected mouse macrophages treated with a novel recom-binant mouse lactoferrinrdquo Annals of Clinical Lab Science vol45 no 5 pp 403ndash410 2015

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

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Disease Markers

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BioMed Research International

OncologyJournal of

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The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

6 Mediators of Inflammation

BALB

cC5

7BL

6TDM alone TDM + LFNaiumlve

TDM alone TDM + LFNaiumlve

Figure 4 Granuloma response to TDM challenge in oral recombinant human lactoferrin-treated mice Two strains of mice (BALBc andC57BL6) were challenged with 25120583g TDM in a water-in-oil emulsion as described [38 101] Human lactoferrin (recombinant 1mg) wasgiven orally at 4 and 6 days after TDM challenge Comparisons are made to naıve mice and to mice only administered TDM HampE staining100x Representative sections from mice at 7 days after TDM challenge119873 ge 8 per experimental group

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

This work was supported in part by NIH Grants 1R41-AI117990-01 and R42-AI051050-05The authors acknowledgePharmaReview Corp (Houston TX) for their kind gifts oflactoferrins and thank Marian L Kruzel PhD and Shen-An Hwang PhD for their insights into the research projectThe authors also thank Ron Gadot and Ai Yamazaki for theirtechnical contributions to the research presented

References

[1] P E M Fine ldquoHerd immunity history theory practicerdquo Epide-miologic Reviews vol 15 no 2 pp 265ndash302 1993

[2] L Ramakrishnan ldquoRevisiting the role of the granuloma intuberculosisrdquo Nature Reviews Immunology vol 12 no 5 pp352ndash366 2012

[3] M Silva Miranda A Breiman S Allain F Deknuydt andF Altare ldquoThe tuberculous granuloma an unsuccessful hostdefence mechanism providing a safety shelter for the bacteriardquoClinical amp Developmental Immunology vol 2012 Article ID139127 14 pages 2012

[4] S A Khader J Rangel-Moreno J J Fountain et al ldquoIn amurinetuberculosis model the absence of homeostatic chemokinesdelays granuloma formation and protective immunityrdquo Journalof Immunology vol 183 no 12 pp 8004ndash8014 2009

[5] AMCooper and S A Khader ldquoThe role of cytokines in the ini-tiation expansion and control of cellular immunity to tubercu-losisrdquo Immunological Reviews vol 226 no 1 pp 191ndash204 2008

[6] IMOrme and R J Basaraba ldquoThe formation of the granulomain tuberculosis infectionrdquo Seminars in Immunology vol 26 no6 pp 601ndash609 2014

[7] I M Orme R T Robinson and A M Cooper ldquoThe balancebetween protective and pathogenic immune responses in theTB-infected lungrdquo Nature Immunology vol 16 no 1 pp 57ndash632014

[8] J K Actor M Olsen C Jagannath and R L Hunter ldquoRela-tionship of survival organism containment and granulomaformation in acute murine tuberculosisrdquo Journal of Interferonand Cytokine Research vol 19 no 10 pp 1183ndash1193 1999

[9] J L Flynn and J Chan ldquoWhatrsquos good for the host is good for thebugrdquo Trends in Microbiology vol 13 no 3 pp 98ndash102 2005

[10] D G Russell ldquoMycobacterium tuberculosis and the intimatediscourse of a chronic infectionrdquo Immunological Reviews vol240 no 1 pp 252ndash268 2011

[11] R L Hunter ldquoPathology of post primary tuberculosis of thelung an illustrated critical reviewrdquo Tuberculosis vol 91 no 6pp 497ndash509 2011

[12] R L Hunter J K Actor S-A Hwang V Karev and C Jagan-nath ldquoPathogenesis of post primary tuberculosis immunity andhypersensitivity in the development of cavitiesrdquoAnnals of Clini-cal and Laboratory Science vol 44 no 4 pp 365ndash387 2014

[13] K J Welsh R L Hunter and J K Actor ldquoTrehalose 661015840-dimy-colatemdasha coat to regulate tuberculosis immunopathogenesisrdquoTuberculosis vol 93 supplement pp S3ndashS9 2013

[14] M-J Kim H C Wainwright M Locketz et al ldquoCaseation ofhuman tuberculosis granulomas correlates with elevated hostlipid metabolismrdquo EMBO Molecular Medicine vol 2 no 7 pp258ndash274 2010

[15] J K Actor E Breij R A Wetsel H Hoffmann R L HunterJr and C Jagannath ldquoA role for complement C5 in organ-ism containment and granulomatous response during murine

Mediators of Inflammation 7

tuberculosisrdquo Scandinavian Journal of Immunology vol 53 no5 pp 464ndash474 2001

[16] J K Actor C D Leonard V E Watson et al ldquoCytokinemRNAexpression and serumcortisol evaluation duringmurinelung inflammation induced by Mycobacterium tuberculosisrdquoCombinatorial Chemistry and High Throughput Screening vol3 no 4 pp 343ndash351 2000

[17] C W Borders A Courtney K Ronen et al ldquoRequisite role forcomplement C5 and the C5a receptor in granulomatous res-ponse to mycobacterial glycolipid trehalose 661015840-dimycolaterdquoScandinavian Journal of Immunology vol 62 no 2 pp 123ndash1302005

[18] R L Hunter C Jagannath and J K Actor ldquoPathology of post-primary tuberculosis in humans and mice contradiction oflong-held beliefsrdquo Tuberculosis vol 87 no 4 pp 267ndash278 2007

[19] C Jagannath H Hoffmann E Sepulveda J K Actor R AWetsel and R L Hunter Jr ldquoHypersusceptibility of AJ miceto tuberculosis is in part due to a deficiency of the fifth comple-ment component (C5)rdquo Scandinavian Journal of Immunologyvol 52 no 4 pp 369ndash379 2000

[20] V E Watson L L Hill L B Owen-Schaub et al ldquoApoptosis inMycobacterium tuberculosis infection in mice exhibiting variedimmunopathologyrdquo Journal of Pathology vol 190 no 2 pp 211ndash220 2000

[21] R H Copenhaver E Sepulveda L Y Armitige et al ldquoAmutantof Mycobacterium tuberculosis H37Rv that lacks expressionof antigen 85A is attenuated in mice but retains vaccinogenicpotentialrdquo Infection and Immunity vol 72 no 12 pp 7084ndash7095 2004

[22] S-A Hwang R Arora M L Kruzel and J K Actor ldquoLactofer-rin enhances efficacy of the BCG vaccine comparison betweentwo inbred mice strains (C57BL6 and BALBc)rdquo Tuberculosisvol 89 supplement 1 pp S49ndashS54 2009

[23] S-A Hwang K Wilk M L Kruzel and J K Actor ldquoA novelrecombinant human lactoferrin augments the BCG vaccine andprotects alveolar integrity upon infection with Mycobacteriumtuberculosis in micerdquo Vaccine vol 27 no 23 pp 3026ndash30342009

[24] S-A Hwang K M Wilk M Budnicka et al ldquoLactoferrinenhanced efficacy of the BCG vaccine to generate host protec-tive responses against challenge with virulent Mycobacteriumtuberculosisrdquo Vaccine vol 25 no 37-38 pp 6730ndash6743 2007

[25] S-A Hwang and J K Actor ldquoLactoferrin modulation of BCG-infected dendritic cell functionsrdquo International Immunologyvol 21 no 10 pp 1185ndash1197 2009

[26] S-A Hwang M L Kruzel and J K Actor ldquoLactoferrin aug-ments BCG vaccine efficacy to generate T helper response andsubsequent protection against challenge with virulentMycobac-terium tuberculosisrdquo International Immunopharmacology vol 5no 3 pp 591ndash599 2005

[27] K M Wilk S-A Hwang and J K Actor ldquoLactoferrin mod-ulation of antigen-presenting-cell response to BCG infectionrdquoPostepy Higieny i Medycyny Doswiadczalnej vol 61 pp 277ndash282 2007

[28] S A Hwang M L Kruzel and J K Actor ldquoCHO expressedrecombinant human lactoferrin as an adjuvant for BCGrdquoInternational Journal of Immunopathology and Pharmacologyvol 28 no 4 pp 452ndash468 2015

[29] R L Perez J Roman G W Staton Jr and R L HunterldquoExtravascular coagulation and fibrinolysis in murine lunginflammation induced by the mycobacterial cord factor

trehalose-66rsquo-dimycolaterdquoAmerican Journal of Respiratory andCritical Care Medicine vol 149 no 2 pp 510ndash518 1994

[30] C A Behling R L Perez M R Kidd G W Staton Jr and RL Hunter ldquoInduction of pulmonary granulomas macrophageprocoagulant activity and tumor necrosis factor-alpha by tre-halose glycolipidsrdquo Annals of Clinical and Laboratory Sciencevol 23 no 4 pp 256ndash266 1993

[31] G S Retzinger S C Meredith R L Hunter K Takayama andF J Kezdy ldquoIdentification of the physiologically active state ofthe mycobacterial glycolipid trehalose 661015840-dimycolate and therole of fibrinogen in the biologic activities of trehalose 661015840-dimycolate monolayersrdquo Journal of Immunology vol 129 no 2pp 735ndash744 1982

[32] G S Retzinger S C Meredith K Takayama R L Hunter andF J Kezdy ldquoThe role of surface in the biological activities oftrehalose 661015840-dimycolate Surface properties and developmentof amodel systemrdquoThe Journal of Biological Chemistry vol 256no 15 pp 8208ndash8216 1981

[33] R L Perez J Roman S Roser et al ldquoCytokinemessage and pro-tein expression during lung granuloma formation and resolu-tion induced by the mycobacterial cord factor trehalose-661015840-dimycolaterdquo Journal of Interferon amp Cytokine Research vol 20no 9 pp 795ndash804 2000

[34] R L Hunter M R Olsen C Jagannath and J K Actor ldquoMult-iple roles of cord factor in the pathogenesis of primary sec-ondary and cavitary tuberculosis including a revised descrip-tion of the pathology of secondary diseaserdquo Annals of Clinicaland Laboratory Science vol 36 no 4 pp 371ndash386 2006

[35] J K Actor ldquoMycobacterial TDM a coat to modulate post pri-mary pathogenesisrdquoMycobacterial Diseases vol 2 no 1 articlee106 2012

[36] R L HunterMOlsen C Jagannath and J K Actor ldquoTrehalose661015840-dimycolate and lipid in the pathogenesis of caseatinggranulomas of tuberculosis in micerdquo The American Journal ofPathology vol 168 no 4 pp 1249ndash1261 2006

[37] A N Abbott T V Guidry K J Welsh et al ldquo11Beta-hydroxy-steroid dehydrogenases are regulated during the pulmonarygranulomatous response to the mycobacterial glycolipidtrehalose-661015840-dimycolaterdquo NeuroImmunoModulation vol 16no 3 pp 147ndash154 2009

[38] K J Welsh A N Abbott S-A Hwang et al ldquoA role for tumournecrosis factor-120572 complement C5 and interleukin-6 in theinitiation and development of the mycobacterial cord factortrehalose 661015840-dimycolate induced granulomatous responserdquoMicrobiology vol 154 no 6 pp 1813ndash1824 2008

[39] A Bekierkunst and E Yarkoni ldquoGranulomatous hypersensitiv-ity to trehalose-66rsquo-dimycolate (cord factor) in mice infectedwith BCGrdquo Infection and Immunity vol 7 no 4 pp 631ndash6381973

[40] H Yamagami TMatsumoto N Fujiwara et al ldquoTrehalose 661015840-dimycolate (cord factor) ofMycobacterium tuberculosis inducesforeign-body- and hypersensitivity-type granulomas in micerdquoInfection and Immunity vol 69 no 2 pp 810ndash815 2001

[41] T VGuidry R LHunter Jr and J K Actor ldquoMycobacterial gly-colipid trehalose 661015840-dimycolate-induced hypersensitive gran-ulomas contribution of CD4+ lymphocytesrdquoMicrobiology vol153 no 10 pp 3360ndash3369 2007

[42] E Rhoades F-F Hsu J B Torrelles J Turk D Chatterjee andD G Russell ldquoIdentification and macrophage-activating activ-ity of glycolipids released from intracellular Mycobacteriumbovis BCGrdquoMolecular Microbiology vol 48 no 4 pp 875ndash8882003

8 Mediators of Inflammation

[43] MMHossain andM-N Norazmi ldquoPattern recognition recep-tors and cytokines in Mycobacterium tuberculosis infectionmdashthe double-edged swordrdquo BioMed Research International vol2013 Article ID 179174 18 pages 2013

[44] D M E Bowdish K Sakamoto M-J Kim et al ldquoMARCOTLR2 and CD14 are required for macrophage cytokine res-ponses to mycobacterial trehalose dimycolate and Mycobac-terium tuberculosisrdquo PLoS Pathogens vol 5 no 6 Article IDe1000474 2009

[45] E Mortaz I M Adcock P Tabarsi et al ldquoInteraction of patternrecognition receptors withMycobacterium tuberculosisrdquo Journalof Clinical Immunology vol 35 no 1 pp 1ndash10 2015

[46] D G Russell ldquoWho puts the tubercle in tuberculosisrdquo NatureReviews Microbiology vol 5 no 1 pp 39ndash47 2007

[47] H Schoenen B Bodendorfer K Hitchens et al ldquoCutting Edgemincle is essential for recognition and adjuvanticity of themycobacterial cord factor and its synthetic analog trehalose-dibehenaterdquo Journal of Immunology vol 184 no 6 pp 2756ndash2760 2010

[48] E Ishikawa T Ishikawa Y S Morita et al ldquoDirect recognitionof themycobacterial glycolipid trehalose dimycolate by C-typelectinMinclerdquo Journal of ExperimentalMedicine vol 206 no 13pp 2879ndash2888 2009

[49] I Matsunaga and D B Moody ldquoMincle is a long soughtreceptor for mycobacterial cord factorrdquo Journal of ExperimentalMedicine vol 206 no 13 pp 2865ndash2868 2009

[50] K Werninghaus A Babiak O Gross et al ldquoAdjuvanticity ofa synthetic cord factor analogue for subunit Mycobacteriumtuberculosis vaccination requires FclR120574-Syk-Card9-dependentinnate immune activationrdquo Journal of Experimental Medicinevol 206 no 1 pp 89ndash97 2009

[51] S LeibundGut-LandmannOGrossM J Robinson et al ldquoSyk-and CARD9-dependent coupling of innate immunity to theinduction of T helper cells that produce interleukin 17rdquo NatureImmunology vol 8 no 6 pp 630ndash638 2007

[52] J Indrigo R L Hunter Jr and J K Actor ldquoInfluence of tre-halose 661015840-dimycolate (TDM) during mycobacterial infectionof bone marrow macrophagesrdquoMicrobiology vol 148 no 7 pp1991ndash1998 2002

[53] J Indrigo R L Hunter Jr and J K Actor ldquoCord factortrehalose 661015840-dimycolate (TDM) mediates trafficking eventsduringmycobacterial infection ofmurinemacrophagesrdquoMicro-biology vol 149 no 8 pp 2049ndash2059 2003

[54] J K Actor M Olsen R L Hunter Jr and Y-J Geng ldquoDys-regulated response to mycobacterial cord factor trehalose-661015840-dimycolate inCD1Dminusminusmicerdquo Journal of Interferon andCytokineResearch vol 21 no 12 pp 1089ndash1096 2001

[55] J K Actor J Indrigo C M Beachdel et al ldquoMycobacterial gly-colipid cord factor trehalose 661015840-dimycolate causes a decreasein serum cortisol during the granulomatous responserdquo Neuro-ImmunoModulation vol 10 no 5 pp 270ndash282 2002

[56] A N Abbott K J Welsh S-A Hwang et al ldquoIL-6 mediates11120573hsd type 2 to effect progression of the mycobacterial cordfactor trehalose 661015840-dimycolate-induced granulomatous res-ponserdquo NeuroImmunoModulation vol 18 no 4 pp 212ndash2252011

[57] T V Guidry M Olsen K-S Kil R L Hunter Jr Y-J Gengand J K Actor ldquoFailure of CD1Dminusminusmice to elicit hypersensitivegranulomas to mycobacterial cord factor trehalose 661015840-dimy-colaterdquo Journal of Interferon amp Cytokine Research vol 24 no 6pp 362ndash371 2004

[58] TVGuidry R LHunter Jr and J KActor ldquoCD3+ cells transferthe hypersensitive granulomatous response to mycobacterialglycolipid trehalose 661015840-dimycolate in micerdquoMicrobiology vol152 no 12 pp 3765ndash3775 2006

[59] B Lonnerdal and S Iyer ldquoLactoferrin molecular structure andbiological functionrdquoAnnual Review of Nutrition vol 15 pp 93ndash110 1995

[60] L Sanchez M Calvo and J H Brock ldquoBiological role of lacto-ferrinrdquo Archives of Disease in Childhood vol 67 no 5 pp 657ndash661 1992

[61] P Manzoni M Rinaldi S Cattani et al ldquoBovine lactoferrinsupplementation for prevention of late-onset sepsis in verylow-birth-weight neonates a randomized trialrdquo Journal of theAmerican Medical Association vol 302 no 13 pp 1421ndash14282009

[62] S Baveye E Elass J Mazurier G Spik and D Legrand ldquoLacto-ferrin a multifunctional glycoprotein involved in the modula-tion of the inflammatory processrdquoClinical Chemistry and Labo-ratory Medicine vol 37 no 3 pp 281ndash286 1999

[63] R D Baynes and W R Bezwoda ldquoLactoferrin and the inflam-matory responserdquo Advances in Experimental Medicine andBiology vol 357 pp 133ndash141 1994

[64] M Kocieba M Zimecki M Kruzel and J Actor ldquoThe adjuvantactivity of lactoferrin in the generation of DTH to ovalbumincan be inhibited by bovine serum albumin bearing alpha-D-mannopyranosyl residuesrdquoCellularampMolecular Biology Lettersvol 7 no 4 pp 1131ndash1136 2002

[65] C G Turin A Zea-Vera A Pezo et al ldquoLactoferrin for pre-vention of neonatal sepsisrdquo Biometals vol 27 no 5 pp 1007ndash1016 2014

[66] T Siqueiros-Cendon S Arevalo-Gallegos B F Iglesias-Figueroa I A Garcıa-Montoya J Salazar-Martınez and QRascon-Cruz ldquoImmunomodulatory effects of lactoferrinrdquo ActaPharmacologica Sinica vol 35 no 5 pp 557ndash566 2014

[67] D Latorre F Berlutti P Valenti S Gessani and P Puddu ldquoLFimmunomodulatory strategies mastering bacterial endotoxinrdquoBiochemistry and Cell Biology vol 90 no 3 pp 269ndash278 2012

[68] D Legrand ldquoLactoferrin a key molecule in immune andinflammatory processesrdquo Biochemistry and Cell Biology vol 90no 3 pp 252ndash268 2012

[69] S-A Hwang K M Wilk Y A Bangale M L Kruzel and JK Actor ldquoLactoferrin modulation of IL-12 and IL-10 responsefrom activated murine leukocytesrdquo Medical Microbiology andImmunology vol 196 no 3 pp 171ndash180 2007

[70] M Zimecki J Mazurier G Spik and J A Kapp ldquoHuman lacto-ferrin induces phenotypic and functional changes in murinesplenic B cellsrdquo Immunology vol 86 no 1 pp 122ndash127 1995

[71] M Zimecki J Artym G Chodaczek et al ldquoImmunoregulatoryfunction of lactoferrin in immunosuppressed and autoimmuneanimalsrdquo Postepy Higieny i Medycyny Doswiadczalnej vol 61pp 283ndash287 2007

[72] M Zimecki and M L Kruzel ldquoMilk-derived proteins andpeptides of potential therapeutic and nutritive valuerdquo Journal ofExperimental Therapeutics and Oncology vol 6 no 2 pp 89ndash106 2007

[73] M Zimecki RMiędzybrodzki J Mazurier andG Spik ldquoRegu-latory effects of lactoferrin and lipopolysaccharide on LFA-1 expression on human peripheral blood mononuclear cellsrdquoArchivum Immunologiae et Therapiae Experimentalis vol 47no 4 pp 257ndash264 1999

Mediators of Inflammation 9

[74] M L Kruzel and M Zimecki ldquoLactoferrin and immunologicdissonance clinical implicationsrdquo Archivum Immunologiae etTherapiae Experimentalis vol 50 no 6 pp 399ndash410 2002

[75] W I Weis M E Taylor and K Drickamer ldquoThe C-type lectinsuperfamily in the immune systemrdquo Immunological Reviewsvol 163 pp 19ndash34 1998

[76] W I Weis K Drickamer and W A Hendrickson ldquoStructureof a C-type mannose-binding protein complexed with an oli-gosacchariderdquo Nature vol 360 no 6400 pp 127ndash134 1992

[77] T Zagulski Z Jarzabek A Zagulska M Jaszczak I EKochanowska and M Zimecki ldquoLactoferrin stimulates killingand clearance of bacteria but does not prevent mortality of dia-betic micerdquo Archivum Immunologiae et Therapiae Experimen-talis vol 49 no 6 pp 431ndash438 2001

[78] M Kruzel Y Harai C Chen and A Castro ldquoRole of lactoferrinin development of systemic inflammatory response syndrome(SIRS)rdquo in Proceedings of the 2nd International ConferenceProgress in Intensive Care Medicine pp 11ndash12 Wroclaw Poland1998

[79] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoThe guta key metabolic organ protected by lactoferrin during experi-mental systemic inflammation in micerdquo in Advances in Lacto-ferrin Research vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Springer New York NY USA 1998

[80] MKruzel T Zagulski andMZimecki ldquoLactoferrin and insult-induced metabolic imbalance in humans and other animalsrdquoin Lactoferrin Structure Function and Applications Proceedingsof the 4th International Conference on Lactoferrin StructureFunction and Applications held in Sapporo Japan 18ndash22 May1999 pp 301ndash311 ICS Elsevier New York NY USA 2000

[81] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoLactofer-rin protects gut mucosal integrity during endotoxemia inducedby lipopolysaccharide in micerdquo Inflammation vol 24 no 1 pp33ndash44 2000

[82] E Zaczynska M Kocięba E Sliwinska and M ZimeckildquoBovine lactoferrin enhances proliferation of human peripheralblood lymphocytes and induces cytokine production in wholeblood culturesrdquo Advances in Clinical and Experimental Medi-cine vol 23 pp 871ndash876 2014

[83] Z Wei T Nishimura and S Yoshida ldquoCharacterization ofglycans in a lactoferrin isoform lactoferrin-ardquo Journal of DairyScience vol 84 no 12 pp 2584ndash2590 2001

[84] B-K Choi J K Actor S Rios et al ldquoRecombinant humanlactoferrin expressed in glycoengineered Pichia pastoris effectof terminal N-acetylneuraminic acid on in vitro secondaryhumoral immune responserdquoGlycoconjugate Journal vol 25 no6 pp 581ndash593 2008

[85] B Samyn-Petit J-PWajdaDubos F Chirat et al ldquoComparativeanalysis of the site-specific N-glycosylation of human lactofer-rin produced in maize and tobacco plantsrdquo European Journal ofBiochemistry vol 270 no 15 pp 3235ndash3242 2003

[86] M Zimecki J Artym M Kocięba M Duk and M L KruzelldquoThe effect of carbohydrate moiety structure on the immu-noregulatory activity of lactoferrin in vitrordquo Cellular ampMolecu-lar Biology Letters vol 19 no 2 pp 284ndash296 2014

[87] J K Actor S-A Hwang and M L Kruzel ldquoLactoferrin as anatural immune modulatorrdquo Current Pharmaceutical Designvol 15 no 17 pp 1956ndash1973 2009

[88] M L Kruzel J K Actor I Boldogh and M ZimeckildquoLactoferrin in health and diseaserdquo Postepy Higieny i MedycynyDoswiadczalnej vol 61 pp 261ndash267 2007

[89] S Baveye E Elass D G Fernig C Blanquart J Mazurier andD Legrand ldquoHuman lactoferrin interacts with soluble CD14and inhibits expression of endothelial adhesion molecules E-selectin and ICAM-1 induced by the CD14-lipopolysaccharidecomplexrdquo Infection and Immunity vol 68 no 12 pp 6519ndash65252000

[90] C S Curran K P Demick and J M Mansfield ldquoLactoferrinactivates macrophages via TLR4-dependent and -independentsignaling pathwaysrdquo Cellular Immunology vol 242 no 1 pp23ndash30 2006

[91] P Puddu D Latorre M Carollo et al ldquoBovine lactoferrincounteracts Toll-Like receptor mediated activation signals inantigen presenting cellsrdquo PLoS ONE vol 6 no 7 Article IDe22504 2011

[92] T M Doherty and M Arditi ldquoTB or not TB That is the ques-tionmdashdoes TLR signaling hold the answerrdquoThe Journal of Clin-ical Investigation vol 114 no 12 pp 1699ndash1703 2004

[93] M A Rahman P Sobia N Gupta L Van Kaer and GDas ldquoMycobacterium tuberculosis subverts the TLR-2mdashMyD88pathway to facilitate its translocation into the cytosolrdquo PLoSONE vol 9 no 1 Article ID e86886 2014

[94] J ArtymM Zimecki andM L Kruzel ldquoEnhanced clearance ofEscherichia coli and Staphylococcus aureus in mice treated withcyclophosphamide and lactoferrinrdquo International Immunophar-macology vol 4 no 9 pp 1149ndash1157 2004

[95] G Chodaczek A Saavedra-Molina A Bacsi M L Kruzel SSur and I Boldogh ldquoIron-mediated dismutation of superoxideanion augments antigen-induced allergic inflammation effectof lactoferrinrdquo Postepy Higieny i Medycyny Doswiadczalnej vol61 pp 268ndash276 2007

[96] M Kruzel Y Harai C Chen and A Castro ldquoThe Gut a keymetabolic organ protected by lactoferrin during experimentalsystemic inflammation in micerdquo in Advances in LactoferrinResearch G Spik D Legrand J Mazurier A Pierce and J-PPerraudin Eds vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Plenum Press 1998

[97] M L Kruzel A Bacsi B Choudhury S Sur and I BoldoghldquoLactoferrin decreases pollen antigen-induced allergic airwayinflammation in a murine model of asthmardquo Immunology vol119 no 2 pp 159ndash166 2006

[98] J K Actor S-A Hwang M Olsen M Zimecki R L HunterJr and M L Kruzel ldquoLactoferrin immunomodulation of DTHresponse in micerdquo International Immunopharmacology vol 2no 4 pp 475ndash486 2002

[99] S-A Hwang M L Kruzel and J K Actor ldquoInfluence of bovinelactoferrin on expression of presentation molecules on BCG-infected bonemarrow derivedmacrophagesrdquo Biochimie vol 91no 1 pp 76ndash85 2009

[100] M Zimecki J Mazurier M Machnicki Z Wieczorek JMontreuil and G Spik ldquoImmunostimulatory activity of lacto-transferrin andmaturation of CD4minus CD8minusmurine thymocytesrdquoImmunology Letters vol 30 no 1 pp 119ndash123 1991

[101] K J Welsh S-A Hwang R L Hunter M L Kruzel and JK Actor ldquoLactoferrin modulation of mycobacterial cord factortrehalose 6-6rsquo-dimycolate induced granulomatous responserdquoTranslational Research vol 156 no 4 pp 207ndash215 2010

[102] M L Bourigault N Segueni S Rose et al ldquoRelative contri-bution of IL-1alpha IL-1beta and TNF to the host responseto Mycobacterium tuberculosis and attenuated M bovis BCGrdquoImmunity Inflammation and Disease vol 1 no 1 pp 47ndash622013

10 Mediators of Inflammation

[103] C Holscher R A Atkinson B Arendse et al ldquoA protective andagonistic function of IL-12p40 in mycobacterial infectionrdquoTheJournal of Immunology vol 167 no 12 pp 6957ndash6966 2001

[104] M Jacobs L Fick N Allie N Brown and B Ryffel ldquoEnhancedimmune response in Mycobacterium bovis bacille calmetteguerin (BCG)-infected IL-10-deficientmicerdquoClinical Chemistryand Laboratory Medicine vol 40 no 9 pp 893ndash902 2002

[105] K J Welsh S-A Hwang S Boyd M L Kruzel R L Hunterand J K Actor ldquoInfluence of oral lactoferrin onMycobacteriumtuberculosis induced immunopathologyrdquo Tuberculosis vol 91supplement 1 pp S105ndashS113 2011

[106] M L Kruzel J K Actor M Zimecki et al ldquoNovel recombinanthuman lactoferrin differential activation of oxidative stressrelated gene expressionrdquo Journal of Biotechnology vol 168 no4 pp 666ndash675 2013

[107] Y Shibata J Gabbard M Yamashita et al ldquoHeat-killedBCG induces biphasic cyclooxygenase 2+ splenic macrophageformationmdashrole of IL-10 and bone marrow precursorsrdquo Journalof Leukocyte Biology vol 80 no 3 pp 590ndash598 2006

[108] H Yamada E Kuroda S Matsumoto T Matsumoto TYamada and U Yamashita ldquoProstaglandin E

2down-regulates

viable Bacille Calmette-Guerin-induced macrophage cytotoxi-city against murine bladder cancer cell MBT-2 in vitrordquoClinicaland Experimental Immunology vol 128 no 1 pp 52ndash58 2002

[109] S Hwang M L Kruzel and J K Actor ldquoEffects of CHO-expressed recombinant lactoferrins on mouse dendritic cellpresentation and functionrdquo Innate Immunity vol 21 no 5 pp553ndash561 2015

[110] K M OShea S-A Hwang and J K Actor ldquoImmune activity ofbcg infected mouse macrophages treated with a novel recom-binant mouse lactoferrinrdquo Annals of Clinical Lab Science vol45 no 5 pp 403ndash410 2015

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

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BioMed Research International

OncologyJournal of

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Oxidative Medicine and Cellular Longevity

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PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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ObesityJournal of

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

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Diabetes ResearchJournal of

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Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Mediators of Inflammation 7

tuberculosisrdquo Scandinavian Journal of Immunology vol 53 no5 pp 464ndash474 2001

[16] J K Actor C D Leonard V E Watson et al ldquoCytokinemRNAexpression and serumcortisol evaluation duringmurinelung inflammation induced by Mycobacterium tuberculosisrdquoCombinatorial Chemistry and High Throughput Screening vol3 no 4 pp 343ndash351 2000

[17] C W Borders A Courtney K Ronen et al ldquoRequisite role forcomplement C5 and the C5a receptor in granulomatous res-ponse to mycobacterial glycolipid trehalose 661015840-dimycolaterdquoScandinavian Journal of Immunology vol 62 no 2 pp 123ndash1302005

[18] R L Hunter C Jagannath and J K Actor ldquoPathology of post-primary tuberculosis in humans and mice contradiction oflong-held beliefsrdquo Tuberculosis vol 87 no 4 pp 267ndash278 2007

[19] C Jagannath H Hoffmann E Sepulveda J K Actor R AWetsel and R L Hunter Jr ldquoHypersusceptibility of AJ miceto tuberculosis is in part due to a deficiency of the fifth comple-ment component (C5)rdquo Scandinavian Journal of Immunologyvol 52 no 4 pp 369ndash379 2000

[20] V E Watson L L Hill L B Owen-Schaub et al ldquoApoptosis inMycobacterium tuberculosis infection in mice exhibiting variedimmunopathologyrdquo Journal of Pathology vol 190 no 2 pp 211ndash220 2000

[21] R H Copenhaver E Sepulveda L Y Armitige et al ldquoAmutantof Mycobacterium tuberculosis H37Rv that lacks expressionof antigen 85A is attenuated in mice but retains vaccinogenicpotentialrdquo Infection and Immunity vol 72 no 12 pp 7084ndash7095 2004

[22] S-A Hwang R Arora M L Kruzel and J K Actor ldquoLactofer-rin enhances efficacy of the BCG vaccine comparison betweentwo inbred mice strains (C57BL6 and BALBc)rdquo Tuberculosisvol 89 supplement 1 pp S49ndashS54 2009

[23] S-A Hwang K Wilk M L Kruzel and J K Actor ldquoA novelrecombinant human lactoferrin augments the BCG vaccine andprotects alveolar integrity upon infection with Mycobacteriumtuberculosis in micerdquo Vaccine vol 27 no 23 pp 3026ndash30342009

[24] S-A Hwang K M Wilk M Budnicka et al ldquoLactoferrinenhanced efficacy of the BCG vaccine to generate host protec-tive responses against challenge with virulent Mycobacteriumtuberculosisrdquo Vaccine vol 25 no 37-38 pp 6730ndash6743 2007

[25] S-A Hwang and J K Actor ldquoLactoferrin modulation of BCG-infected dendritic cell functionsrdquo International Immunologyvol 21 no 10 pp 1185ndash1197 2009

[26] S-A Hwang M L Kruzel and J K Actor ldquoLactoferrin aug-ments BCG vaccine efficacy to generate T helper response andsubsequent protection against challenge with virulentMycobac-terium tuberculosisrdquo International Immunopharmacology vol 5no 3 pp 591ndash599 2005

[27] K M Wilk S-A Hwang and J K Actor ldquoLactoferrin mod-ulation of antigen-presenting-cell response to BCG infectionrdquoPostepy Higieny i Medycyny Doswiadczalnej vol 61 pp 277ndash282 2007

[28] S A Hwang M L Kruzel and J K Actor ldquoCHO expressedrecombinant human lactoferrin as an adjuvant for BCGrdquoInternational Journal of Immunopathology and Pharmacologyvol 28 no 4 pp 452ndash468 2015

[29] R L Perez J Roman G W Staton Jr and R L HunterldquoExtravascular coagulation and fibrinolysis in murine lunginflammation induced by the mycobacterial cord factor

trehalose-66rsquo-dimycolaterdquoAmerican Journal of Respiratory andCritical Care Medicine vol 149 no 2 pp 510ndash518 1994

[30] C A Behling R L Perez M R Kidd G W Staton Jr and RL Hunter ldquoInduction of pulmonary granulomas macrophageprocoagulant activity and tumor necrosis factor-alpha by tre-halose glycolipidsrdquo Annals of Clinical and Laboratory Sciencevol 23 no 4 pp 256ndash266 1993

[31] G S Retzinger S C Meredith R L Hunter K Takayama andF J Kezdy ldquoIdentification of the physiologically active state ofthe mycobacterial glycolipid trehalose 661015840-dimycolate and therole of fibrinogen in the biologic activities of trehalose 661015840-dimycolate monolayersrdquo Journal of Immunology vol 129 no 2pp 735ndash744 1982

[32] G S Retzinger S C Meredith K Takayama R L Hunter andF J Kezdy ldquoThe role of surface in the biological activities oftrehalose 661015840-dimycolate Surface properties and developmentof amodel systemrdquoThe Journal of Biological Chemistry vol 256no 15 pp 8208ndash8216 1981

[33] R L Perez J Roman S Roser et al ldquoCytokinemessage and pro-tein expression during lung granuloma formation and resolu-tion induced by the mycobacterial cord factor trehalose-661015840-dimycolaterdquo Journal of Interferon amp Cytokine Research vol 20no 9 pp 795ndash804 2000

[34] R L Hunter M R Olsen C Jagannath and J K Actor ldquoMult-iple roles of cord factor in the pathogenesis of primary sec-ondary and cavitary tuberculosis including a revised descrip-tion of the pathology of secondary diseaserdquo Annals of Clinicaland Laboratory Science vol 36 no 4 pp 371ndash386 2006

[35] J K Actor ldquoMycobacterial TDM a coat to modulate post pri-mary pathogenesisrdquoMycobacterial Diseases vol 2 no 1 articlee106 2012

[36] R L HunterMOlsen C Jagannath and J K Actor ldquoTrehalose661015840-dimycolate and lipid in the pathogenesis of caseatinggranulomas of tuberculosis in micerdquo The American Journal ofPathology vol 168 no 4 pp 1249ndash1261 2006

[37] A N Abbott T V Guidry K J Welsh et al ldquo11Beta-hydroxy-steroid dehydrogenases are regulated during the pulmonarygranulomatous response to the mycobacterial glycolipidtrehalose-661015840-dimycolaterdquo NeuroImmunoModulation vol 16no 3 pp 147ndash154 2009

[38] K J Welsh A N Abbott S-A Hwang et al ldquoA role for tumournecrosis factor-120572 complement C5 and interleukin-6 in theinitiation and development of the mycobacterial cord factortrehalose 661015840-dimycolate induced granulomatous responserdquoMicrobiology vol 154 no 6 pp 1813ndash1824 2008

[39] A Bekierkunst and E Yarkoni ldquoGranulomatous hypersensitiv-ity to trehalose-66rsquo-dimycolate (cord factor) in mice infectedwith BCGrdquo Infection and Immunity vol 7 no 4 pp 631ndash6381973

[40] H Yamagami TMatsumoto N Fujiwara et al ldquoTrehalose 661015840-dimycolate (cord factor) ofMycobacterium tuberculosis inducesforeign-body- and hypersensitivity-type granulomas in micerdquoInfection and Immunity vol 69 no 2 pp 810ndash815 2001

[41] T VGuidry R LHunter Jr and J K Actor ldquoMycobacterial gly-colipid trehalose 661015840-dimycolate-induced hypersensitive gran-ulomas contribution of CD4+ lymphocytesrdquoMicrobiology vol153 no 10 pp 3360ndash3369 2007

[42] E Rhoades F-F Hsu J B Torrelles J Turk D Chatterjee andD G Russell ldquoIdentification and macrophage-activating activ-ity of glycolipids released from intracellular Mycobacteriumbovis BCGrdquoMolecular Microbiology vol 48 no 4 pp 875ndash8882003

8 Mediators of Inflammation

[43] MMHossain andM-N Norazmi ldquoPattern recognition recep-tors and cytokines in Mycobacterium tuberculosis infectionmdashthe double-edged swordrdquo BioMed Research International vol2013 Article ID 179174 18 pages 2013

[44] D M E Bowdish K Sakamoto M-J Kim et al ldquoMARCOTLR2 and CD14 are required for macrophage cytokine res-ponses to mycobacterial trehalose dimycolate and Mycobac-terium tuberculosisrdquo PLoS Pathogens vol 5 no 6 Article IDe1000474 2009

[45] E Mortaz I M Adcock P Tabarsi et al ldquoInteraction of patternrecognition receptors withMycobacterium tuberculosisrdquo Journalof Clinical Immunology vol 35 no 1 pp 1ndash10 2015

[46] D G Russell ldquoWho puts the tubercle in tuberculosisrdquo NatureReviews Microbiology vol 5 no 1 pp 39ndash47 2007

[47] H Schoenen B Bodendorfer K Hitchens et al ldquoCutting Edgemincle is essential for recognition and adjuvanticity of themycobacterial cord factor and its synthetic analog trehalose-dibehenaterdquo Journal of Immunology vol 184 no 6 pp 2756ndash2760 2010

[48] E Ishikawa T Ishikawa Y S Morita et al ldquoDirect recognitionof themycobacterial glycolipid trehalose dimycolate by C-typelectinMinclerdquo Journal of ExperimentalMedicine vol 206 no 13pp 2879ndash2888 2009

[49] I Matsunaga and D B Moody ldquoMincle is a long soughtreceptor for mycobacterial cord factorrdquo Journal of ExperimentalMedicine vol 206 no 13 pp 2865ndash2868 2009

[50] K Werninghaus A Babiak O Gross et al ldquoAdjuvanticity ofa synthetic cord factor analogue for subunit Mycobacteriumtuberculosis vaccination requires FclR120574-Syk-Card9-dependentinnate immune activationrdquo Journal of Experimental Medicinevol 206 no 1 pp 89ndash97 2009

[51] S LeibundGut-LandmannOGrossM J Robinson et al ldquoSyk-and CARD9-dependent coupling of innate immunity to theinduction of T helper cells that produce interleukin 17rdquo NatureImmunology vol 8 no 6 pp 630ndash638 2007

[52] J Indrigo R L Hunter Jr and J K Actor ldquoInfluence of tre-halose 661015840-dimycolate (TDM) during mycobacterial infectionof bone marrow macrophagesrdquoMicrobiology vol 148 no 7 pp1991ndash1998 2002

[53] J Indrigo R L Hunter Jr and J K Actor ldquoCord factortrehalose 661015840-dimycolate (TDM) mediates trafficking eventsduringmycobacterial infection ofmurinemacrophagesrdquoMicro-biology vol 149 no 8 pp 2049ndash2059 2003

[54] J K Actor M Olsen R L Hunter Jr and Y-J Geng ldquoDys-regulated response to mycobacterial cord factor trehalose-661015840-dimycolate inCD1Dminusminusmicerdquo Journal of Interferon andCytokineResearch vol 21 no 12 pp 1089ndash1096 2001

[55] J K Actor J Indrigo C M Beachdel et al ldquoMycobacterial gly-colipid cord factor trehalose 661015840-dimycolate causes a decreasein serum cortisol during the granulomatous responserdquo Neuro-ImmunoModulation vol 10 no 5 pp 270ndash282 2002

[56] A N Abbott K J Welsh S-A Hwang et al ldquoIL-6 mediates11120573hsd type 2 to effect progression of the mycobacterial cordfactor trehalose 661015840-dimycolate-induced granulomatous res-ponserdquo NeuroImmunoModulation vol 18 no 4 pp 212ndash2252011

[57] T V Guidry M Olsen K-S Kil R L Hunter Jr Y-J Gengand J K Actor ldquoFailure of CD1Dminusminusmice to elicit hypersensitivegranulomas to mycobacterial cord factor trehalose 661015840-dimy-colaterdquo Journal of Interferon amp Cytokine Research vol 24 no 6pp 362ndash371 2004

[58] TVGuidry R LHunter Jr and J KActor ldquoCD3+ cells transferthe hypersensitive granulomatous response to mycobacterialglycolipid trehalose 661015840-dimycolate in micerdquoMicrobiology vol152 no 12 pp 3765ndash3775 2006

[59] B Lonnerdal and S Iyer ldquoLactoferrin molecular structure andbiological functionrdquoAnnual Review of Nutrition vol 15 pp 93ndash110 1995

[60] L Sanchez M Calvo and J H Brock ldquoBiological role of lacto-ferrinrdquo Archives of Disease in Childhood vol 67 no 5 pp 657ndash661 1992

[61] P Manzoni M Rinaldi S Cattani et al ldquoBovine lactoferrinsupplementation for prevention of late-onset sepsis in verylow-birth-weight neonates a randomized trialrdquo Journal of theAmerican Medical Association vol 302 no 13 pp 1421ndash14282009

[62] S Baveye E Elass J Mazurier G Spik and D Legrand ldquoLacto-ferrin a multifunctional glycoprotein involved in the modula-tion of the inflammatory processrdquoClinical Chemistry and Labo-ratory Medicine vol 37 no 3 pp 281ndash286 1999

[63] R D Baynes and W R Bezwoda ldquoLactoferrin and the inflam-matory responserdquo Advances in Experimental Medicine andBiology vol 357 pp 133ndash141 1994

[64] M Kocieba M Zimecki M Kruzel and J Actor ldquoThe adjuvantactivity of lactoferrin in the generation of DTH to ovalbumincan be inhibited by bovine serum albumin bearing alpha-D-mannopyranosyl residuesrdquoCellularampMolecular Biology Lettersvol 7 no 4 pp 1131ndash1136 2002

[65] C G Turin A Zea-Vera A Pezo et al ldquoLactoferrin for pre-vention of neonatal sepsisrdquo Biometals vol 27 no 5 pp 1007ndash1016 2014

[66] T Siqueiros-Cendon S Arevalo-Gallegos B F Iglesias-Figueroa I A Garcıa-Montoya J Salazar-Martınez and QRascon-Cruz ldquoImmunomodulatory effects of lactoferrinrdquo ActaPharmacologica Sinica vol 35 no 5 pp 557ndash566 2014

[67] D Latorre F Berlutti P Valenti S Gessani and P Puddu ldquoLFimmunomodulatory strategies mastering bacterial endotoxinrdquoBiochemistry and Cell Biology vol 90 no 3 pp 269ndash278 2012

[68] D Legrand ldquoLactoferrin a key molecule in immune andinflammatory processesrdquo Biochemistry and Cell Biology vol 90no 3 pp 252ndash268 2012

[69] S-A Hwang K M Wilk Y A Bangale M L Kruzel and JK Actor ldquoLactoferrin modulation of IL-12 and IL-10 responsefrom activated murine leukocytesrdquo Medical Microbiology andImmunology vol 196 no 3 pp 171ndash180 2007

[70] M Zimecki J Mazurier G Spik and J A Kapp ldquoHuman lacto-ferrin induces phenotypic and functional changes in murinesplenic B cellsrdquo Immunology vol 86 no 1 pp 122ndash127 1995

[71] M Zimecki J Artym G Chodaczek et al ldquoImmunoregulatoryfunction of lactoferrin in immunosuppressed and autoimmuneanimalsrdquo Postepy Higieny i Medycyny Doswiadczalnej vol 61pp 283ndash287 2007

[72] M Zimecki and M L Kruzel ldquoMilk-derived proteins andpeptides of potential therapeutic and nutritive valuerdquo Journal ofExperimental Therapeutics and Oncology vol 6 no 2 pp 89ndash106 2007

[73] M Zimecki RMiędzybrodzki J Mazurier andG Spik ldquoRegu-latory effects of lactoferrin and lipopolysaccharide on LFA-1 expression on human peripheral blood mononuclear cellsrdquoArchivum Immunologiae et Therapiae Experimentalis vol 47no 4 pp 257ndash264 1999

Mediators of Inflammation 9

[74] M L Kruzel and M Zimecki ldquoLactoferrin and immunologicdissonance clinical implicationsrdquo Archivum Immunologiae etTherapiae Experimentalis vol 50 no 6 pp 399ndash410 2002

[75] W I Weis M E Taylor and K Drickamer ldquoThe C-type lectinsuperfamily in the immune systemrdquo Immunological Reviewsvol 163 pp 19ndash34 1998

[76] W I Weis K Drickamer and W A Hendrickson ldquoStructureof a C-type mannose-binding protein complexed with an oli-gosacchariderdquo Nature vol 360 no 6400 pp 127ndash134 1992

[77] T Zagulski Z Jarzabek A Zagulska M Jaszczak I EKochanowska and M Zimecki ldquoLactoferrin stimulates killingand clearance of bacteria but does not prevent mortality of dia-betic micerdquo Archivum Immunologiae et Therapiae Experimen-talis vol 49 no 6 pp 431ndash438 2001

[78] M Kruzel Y Harai C Chen and A Castro ldquoRole of lactoferrinin development of systemic inflammatory response syndrome(SIRS)rdquo in Proceedings of the 2nd International ConferenceProgress in Intensive Care Medicine pp 11ndash12 Wroclaw Poland1998

[79] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoThe guta key metabolic organ protected by lactoferrin during experi-mental systemic inflammation in micerdquo in Advances in Lacto-ferrin Research vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Springer New York NY USA 1998

[80] MKruzel T Zagulski andMZimecki ldquoLactoferrin and insult-induced metabolic imbalance in humans and other animalsrdquoin Lactoferrin Structure Function and Applications Proceedingsof the 4th International Conference on Lactoferrin StructureFunction and Applications held in Sapporo Japan 18ndash22 May1999 pp 301ndash311 ICS Elsevier New York NY USA 2000

[81] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoLactofer-rin protects gut mucosal integrity during endotoxemia inducedby lipopolysaccharide in micerdquo Inflammation vol 24 no 1 pp33ndash44 2000

[82] E Zaczynska M Kocięba E Sliwinska and M ZimeckildquoBovine lactoferrin enhances proliferation of human peripheralblood lymphocytes and induces cytokine production in wholeblood culturesrdquo Advances in Clinical and Experimental Medi-cine vol 23 pp 871ndash876 2014

[83] Z Wei T Nishimura and S Yoshida ldquoCharacterization ofglycans in a lactoferrin isoform lactoferrin-ardquo Journal of DairyScience vol 84 no 12 pp 2584ndash2590 2001

[84] B-K Choi J K Actor S Rios et al ldquoRecombinant humanlactoferrin expressed in glycoengineered Pichia pastoris effectof terminal N-acetylneuraminic acid on in vitro secondaryhumoral immune responserdquoGlycoconjugate Journal vol 25 no6 pp 581ndash593 2008

[85] B Samyn-Petit J-PWajdaDubos F Chirat et al ldquoComparativeanalysis of the site-specific N-glycosylation of human lactofer-rin produced in maize and tobacco plantsrdquo European Journal ofBiochemistry vol 270 no 15 pp 3235ndash3242 2003

[86] M Zimecki J Artym M Kocięba M Duk and M L KruzelldquoThe effect of carbohydrate moiety structure on the immu-noregulatory activity of lactoferrin in vitrordquo Cellular ampMolecu-lar Biology Letters vol 19 no 2 pp 284ndash296 2014

[87] J K Actor S-A Hwang and M L Kruzel ldquoLactoferrin as anatural immune modulatorrdquo Current Pharmaceutical Designvol 15 no 17 pp 1956ndash1973 2009

[88] M L Kruzel J K Actor I Boldogh and M ZimeckildquoLactoferrin in health and diseaserdquo Postepy Higieny i MedycynyDoswiadczalnej vol 61 pp 261ndash267 2007

[89] S Baveye E Elass D G Fernig C Blanquart J Mazurier andD Legrand ldquoHuman lactoferrin interacts with soluble CD14and inhibits expression of endothelial adhesion molecules E-selectin and ICAM-1 induced by the CD14-lipopolysaccharidecomplexrdquo Infection and Immunity vol 68 no 12 pp 6519ndash65252000

[90] C S Curran K P Demick and J M Mansfield ldquoLactoferrinactivates macrophages via TLR4-dependent and -independentsignaling pathwaysrdquo Cellular Immunology vol 242 no 1 pp23ndash30 2006

[91] P Puddu D Latorre M Carollo et al ldquoBovine lactoferrincounteracts Toll-Like receptor mediated activation signals inantigen presenting cellsrdquo PLoS ONE vol 6 no 7 Article IDe22504 2011

[92] T M Doherty and M Arditi ldquoTB or not TB That is the ques-tionmdashdoes TLR signaling hold the answerrdquoThe Journal of Clin-ical Investigation vol 114 no 12 pp 1699ndash1703 2004

[93] M A Rahman P Sobia N Gupta L Van Kaer and GDas ldquoMycobacterium tuberculosis subverts the TLR-2mdashMyD88pathway to facilitate its translocation into the cytosolrdquo PLoSONE vol 9 no 1 Article ID e86886 2014

[94] J ArtymM Zimecki andM L Kruzel ldquoEnhanced clearance ofEscherichia coli and Staphylococcus aureus in mice treated withcyclophosphamide and lactoferrinrdquo International Immunophar-macology vol 4 no 9 pp 1149ndash1157 2004

[95] G Chodaczek A Saavedra-Molina A Bacsi M L Kruzel SSur and I Boldogh ldquoIron-mediated dismutation of superoxideanion augments antigen-induced allergic inflammation effectof lactoferrinrdquo Postepy Higieny i Medycyny Doswiadczalnej vol61 pp 268ndash276 2007

[96] M Kruzel Y Harai C Chen and A Castro ldquoThe Gut a keymetabolic organ protected by lactoferrin during experimentalsystemic inflammation in micerdquo in Advances in LactoferrinResearch G Spik D Legrand J Mazurier A Pierce and J-PPerraudin Eds vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Plenum Press 1998

[97] M L Kruzel A Bacsi B Choudhury S Sur and I BoldoghldquoLactoferrin decreases pollen antigen-induced allergic airwayinflammation in a murine model of asthmardquo Immunology vol119 no 2 pp 159ndash166 2006

[98] J K Actor S-A Hwang M Olsen M Zimecki R L HunterJr and M L Kruzel ldquoLactoferrin immunomodulation of DTHresponse in micerdquo International Immunopharmacology vol 2no 4 pp 475ndash486 2002

[99] S-A Hwang M L Kruzel and J K Actor ldquoInfluence of bovinelactoferrin on expression of presentation molecules on BCG-infected bonemarrow derivedmacrophagesrdquo Biochimie vol 91no 1 pp 76ndash85 2009

[100] M Zimecki J Mazurier M Machnicki Z Wieczorek JMontreuil and G Spik ldquoImmunostimulatory activity of lacto-transferrin andmaturation of CD4minus CD8minusmurine thymocytesrdquoImmunology Letters vol 30 no 1 pp 119ndash123 1991

[101] K J Welsh S-A Hwang R L Hunter M L Kruzel and JK Actor ldquoLactoferrin modulation of mycobacterial cord factortrehalose 6-6rsquo-dimycolate induced granulomatous responserdquoTranslational Research vol 156 no 4 pp 207ndash215 2010

[102] M L Bourigault N Segueni S Rose et al ldquoRelative contri-bution of IL-1alpha IL-1beta and TNF to the host responseto Mycobacterium tuberculosis and attenuated M bovis BCGrdquoImmunity Inflammation and Disease vol 1 no 1 pp 47ndash622013

10 Mediators of Inflammation

[103] C Holscher R A Atkinson B Arendse et al ldquoA protective andagonistic function of IL-12p40 in mycobacterial infectionrdquoTheJournal of Immunology vol 167 no 12 pp 6957ndash6966 2001

[104] M Jacobs L Fick N Allie N Brown and B Ryffel ldquoEnhancedimmune response in Mycobacterium bovis bacille calmetteguerin (BCG)-infected IL-10-deficientmicerdquoClinical Chemistryand Laboratory Medicine vol 40 no 9 pp 893ndash902 2002

[105] K J Welsh S-A Hwang S Boyd M L Kruzel R L Hunterand J K Actor ldquoInfluence of oral lactoferrin onMycobacteriumtuberculosis induced immunopathologyrdquo Tuberculosis vol 91supplement 1 pp S105ndashS113 2011

[106] M L Kruzel J K Actor M Zimecki et al ldquoNovel recombinanthuman lactoferrin differential activation of oxidative stressrelated gene expressionrdquo Journal of Biotechnology vol 168 no4 pp 666ndash675 2013

[107] Y Shibata J Gabbard M Yamashita et al ldquoHeat-killedBCG induces biphasic cyclooxygenase 2+ splenic macrophageformationmdashrole of IL-10 and bone marrow precursorsrdquo Journalof Leukocyte Biology vol 80 no 3 pp 590ndash598 2006

[108] H Yamada E Kuroda S Matsumoto T Matsumoto TYamada and U Yamashita ldquoProstaglandin E

2down-regulates

viable Bacille Calmette-Guerin-induced macrophage cytotoxi-city against murine bladder cancer cell MBT-2 in vitrordquoClinicaland Experimental Immunology vol 128 no 1 pp 52ndash58 2002

[109] S Hwang M L Kruzel and J K Actor ldquoEffects of CHO-expressed recombinant lactoferrins on mouse dendritic cellpresentation and functionrdquo Innate Immunity vol 21 no 5 pp553ndash561 2015

[110] K M OShea S-A Hwang and J K Actor ldquoImmune activity ofbcg infected mouse macrophages treated with a novel recom-binant mouse lactoferrinrdquo Annals of Clinical Lab Science vol45 no 5 pp 403ndash410 2015

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

8 Mediators of Inflammation

[43] MMHossain andM-N Norazmi ldquoPattern recognition recep-tors and cytokines in Mycobacterium tuberculosis infectionmdashthe double-edged swordrdquo BioMed Research International vol2013 Article ID 179174 18 pages 2013

[44] D M E Bowdish K Sakamoto M-J Kim et al ldquoMARCOTLR2 and CD14 are required for macrophage cytokine res-ponses to mycobacterial trehalose dimycolate and Mycobac-terium tuberculosisrdquo PLoS Pathogens vol 5 no 6 Article IDe1000474 2009

[45] E Mortaz I M Adcock P Tabarsi et al ldquoInteraction of patternrecognition receptors withMycobacterium tuberculosisrdquo Journalof Clinical Immunology vol 35 no 1 pp 1ndash10 2015

[46] D G Russell ldquoWho puts the tubercle in tuberculosisrdquo NatureReviews Microbiology vol 5 no 1 pp 39ndash47 2007

[47] H Schoenen B Bodendorfer K Hitchens et al ldquoCutting Edgemincle is essential for recognition and adjuvanticity of themycobacterial cord factor and its synthetic analog trehalose-dibehenaterdquo Journal of Immunology vol 184 no 6 pp 2756ndash2760 2010

[48] E Ishikawa T Ishikawa Y S Morita et al ldquoDirect recognitionof themycobacterial glycolipid trehalose dimycolate by C-typelectinMinclerdquo Journal of ExperimentalMedicine vol 206 no 13pp 2879ndash2888 2009

[49] I Matsunaga and D B Moody ldquoMincle is a long soughtreceptor for mycobacterial cord factorrdquo Journal of ExperimentalMedicine vol 206 no 13 pp 2865ndash2868 2009

[50] K Werninghaus A Babiak O Gross et al ldquoAdjuvanticity ofa synthetic cord factor analogue for subunit Mycobacteriumtuberculosis vaccination requires FclR120574-Syk-Card9-dependentinnate immune activationrdquo Journal of Experimental Medicinevol 206 no 1 pp 89ndash97 2009

[51] S LeibundGut-LandmannOGrossM J Robinson et al ldquoSyk-and CARD9-dependent coupling of innate immunity to theinduction of T helper cells that produce interleukin 17rdquo NatureImmunology vol 8 no 6 pp 630ndash638 2007

[52] J Indrigo R L Hunter Jr and J K Actor ldquoInfluence of tre-halose 661015840-dimycolate (TDM) during mycobacterial infectionof bone marrow macrophagesrdquoMicrobiology vol 148 no 7 pp1991ndash1998 2002

[53] J Indrigo R L Hunter Jr and J K Actor ldquoCord factortrehalose 661015840-dimycolate (TDM) mediates trafficking eventsduringmycobacterial infection ofmurinemacrophagesrdquoMicro-biology vol 149 no 8 pp 2049ndash2059 2003

[54] J K Actor M Olsen R L Hunter Jr and Y-J Geng ldquoDys-regulated response to mycobacterial cord factor trehalose-661015840-dimycolate inCD1Dminusminusmicerdquo Journal of Interferon andCytokineResearch vol 21 no 12 pp 1089ndash1096 2001

[55] J K Actor J Indrigo C M Beachdel et al ldquoMycobacterial gly-colipid cord factor trehalose 661015840-dimycolate causes a decreasein serum cortisol during the granulomatous responserdquo Neuro-ImmunoModulation vol 10 no 5 pp 270ndash282 2002

[56] A N Abbott K J Welsh S-A Hwang et al ldquoIL-6 mediates11120573hsd type 2 to effect progression of the mycobacterial cordfactor trehalose 661015840-dimycolate-induced granulomatous res-ponserdquo NeuroImmunoModulation vol 18 no 4 pp 212ndash2252011

[57] T V Guidry M Olsen K-S Kil R L Hunter Jr Y-J Gengand J K Actor ldquoFailure of CD1Dminusminusmice to elicit hypersensitivegranulomas to mycobacterial cord factor trehalose 661015840-dimy-colaterdquo Journal of Interferon amp Cytokine Research vol 24 no 6pp 362ndash371 2004

[58] TVGuidry R LHunter Jr and J KActor ldquoCD3+ cells transferthe hypersensitive granulomatous response to mycobacterialglycolipid trehalose 661015840-dimycolate in micerdquoMicrobiology vol152 no 12 pp 3765ndash3775 2006

[59] B Lonnerdal and S Iyer ldquoLactoferrin molecular structure andbiological functionrdquoAnnual Review of Nutrition vol 15 pp 93ndash110 1995

[60] L Sanchez M Calvo and J H Brock ldquoBiological role of lacto-ferrinrdquo Archives of Disease in Childhood vol 67 no 5 pp 657ndash661 1992

[61] P Manzoni M Rinaldi S Cattani et al ldquoBovine lactoferrinsupplementation for prevention of late-onset sepsis in verylow-birth-weight neonates a randomized trialrdquo Journal of theAmerican Medical Association vol 302 no 13 pp 1421ndash14282009

[62] S Baveye E Elass J Mazurier G Spik and D Legrand ldquoLacto-ferrin a multifunctional glycoprotein involved in the modula-tion of the inflammatory processrdquoClinical Chemistry and Labo-ratory Medicine vol 37 no 3 pp 281ndash286 1999

[63] R D Baynes and W R Bezwoda ldquoLactoferrin and the inflam-matory responserdquo Advances in Experimental Medicine andBiology vol 357 pp 133ndash141 1994

[64] M Kocieba M Zimecki M Kruzel and J Actor ldquoThe adjuvantactivity of lactoferrin in the generation of DTH to ovalbumincan be inhibited by bovine serum albumin bearing alpha-D-mannopyranosyl residuesrdquoCellularampMolecular Biology Lettersvol 7 no 4 pp 1131ndash1136 2002

[65] C G Turin A Zea-Vera A Pezo et al ldquoLactoferrin for pre-vention of neonatal sepsisrdquo Biometals vol 27 no 5 pp 1007ndash1016 2014

[66] T Siqueiros-Cendon S Arevalo-Gallegos B F Iglesias-Figueroa I A Garcıa-Montoya J Salazar-Martınez and QRascon-Cruz ldquoImmunomodulatory effects of lactoferrinrdquo ActaPharmacologica Sinica vol 35 no 5 pp 557ndash566 2014

[67] D Latorre F Berlutti P Valenti S Gessani and P Puddu ldquoLFimmunomodulatory strategies mastering bacterial endotoxinrdquoBiochemistry and Cell Biology vol 90 no 3 pp 269ndash278 2012

[68] D Legrand ldquoLactoferrin a key molecule in immune andinflammatory processesrdquo Biochemistry and Cell Biology vol 90no 3 pp 252ndash268 2012

[69] S-A Hwang K M Wilk Y A Bangale M L Kruzel and JK Actor ldquoLactoferrin modulation of IL-12 and IL-10 responsefrom activated murine leukocytesrdquo Medical Microbiology andImmunology vol 196 no 3 pp 171ndash180 2007

[70] M Zimecki J Mazurier G Spik and J A Kapp ldquoHuman lacto-ferrin induces phenotypic and functional changes in murinesplenic B cellsrdquo Immunology vol 86 no 1 pp 122ndash127 1995

[71] M Zimecki J Artym G Chodaczek et al ldquoImmunoregulatoryfunction of lactoferrin in immunosuppressed and autoimmuneanimalsrdquo Postepy Higieny i Medycyny Doswiadczalnej vol 61pp 283ndash287 2007

[72] M Zimecki and M L Kruzel ldquoMilk-derived proteins andpeptides of potential therapeutic and nutritive valuerdquo Journal ofExperimental Therapeutics and Oncology vol 6 no 2 pp 89ndash106 2007

[73] M Zimecki RMiędzybrodzki J Mazurier andG Spik ldquoRegu-latory effects of lactoferrin and lipopolysaccharide on LFA-1 expression on human peripheral blood mononuclear cellsrdquoArchivum Immunologiae et Therapiae Experimentalis vol 47no 4 pp 257ndash264 1999

Mediators of Inflammation 9

[74] M L Kruzel and M Zimecki ldquoLactoferrin and immunologicdissonance clinical implicationsrdquo Archivum Immunologiae etTherapiae Experimentalis vol 50 no 6 pp 399ndash410 2002

[75] W I Weis M E Taylor and K Drickamer ldquoThe C-type lectinsuperfamily in the immune systemrdquo Immunological Reviewsvol 163 pp 19ndash34 1998

[76] W I Weis K Drickamer and W A Hendrickson ldquoStructureof a C-type mannose-binding protein complexed with an oli-gosacchariderdquo Nature vol 360 no 6400 pp 127ndash134 1992

[77] T Zagulski Z Jarzabek A Zagulska M Jaszczak I EKochanowska and M Zimecki ldquoLactoferrin stimulates killingand clearance of bacteria but does not prevent mortality of dia-betic micerdquo Archivum Immunologiae et Therapiae Experimen-talis vol 49 no 6 pp 431ndash438 2001

[78] M Kruzel Y Harai C Chen and A Castro ldquoRole of lactoferrinin development of systemic inflammatory response syndrome(SIRS)rdquo in Proceedings of the 2nd International ConferenceProgress in Intensive Care Medicine pp 11ndash12 Wroclaw Poland1998

[79] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoThe guta key metabolic organ protected by lactoferrin during experi-mental systemic inflammation in micerdquo in Advances in Lacto-ferrin Research vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Springer New York NY USA 1998

[80] MKruzel T Zagulski andMZimecki ldquoLactoferrin and insult-induced metabolic imbalance in humans and other animalsrdquoin Lactoferrin Structure Function and Applications Proceedingsof the 4th International Conference on Lactoferrin StructureFunction and Applications held in Sapporo Japan 18ndash22 May1999 pp 301ndash311 ICS Elsevier New York NY USA 2000

[81] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoLactofer-rin protects gut mucosal integrity during endotoxemia inducedby lipopolysaccharide in micerdquo Inflammation vol 24 no 1 pp33ndash44 2000

[82] E Zaczynska M Kocięba E Sliwinska and M ZimeckildquoBovine lactoferrin enhances proliferation of human peripheralblood lymphocytes and induces cytokine production in wholeblood culturesrdquo Advances in Clinical and Experimental Medi-cine vol 23 pp 871ndash876 2014

[83] Z Wei T Nishimura and S Yoshida ldquoCharacterization ofglycans in a lactoferrin isoform lactoferrin-ardquo Journal of DairyScience vol 84 no 12 pp 2584ndash2590 2001

[84] B-K Choi J K Actor S Rios et al ldquoRecombinant humanlactoferrin expressed in glycoengineered Pichia pastoris effectof terminal N-acetylneuraminic acid on in vitro secondaryhumoral immune responserdquoGlycoconjugate Journal vol 25 no6 pp 581ndash593 2008

[85] B Samyn-Petit J-PWajdaDubos F Chirat et al ldquoComparativeanalysis of the site-specific N-glycosylation of human lactofer-rin produced in maize and tobacco plantsrdquo European Journal ofBiochemistry vol 270 no 15 pp 3235ndash3242 2003

[86] M Zimecki J Artym M Kocięba M Duk and M L KruzelldquoThe effect of carbohydrate moiety structure on the immu-noregulatory activity of lactoferrin in vitrordquo Cellular ampMolecu-lar Biology Letters vol 19 no 2 pp 284ndash296 2014

[87] J K Actor S-A Hwang and M L Kruzel ldquoLactoferrin as anatural immune modulatorrdquo Current Pharmaceutical Designvol 15 no 17 pp 1956ndash1973 2009

[88] M L Kruzel J K Actor I Boldogh and M ZimeckildquoLactoferrin in health and diseaserdquo Postepy Higieny i MedycynyDoswiadczalnej vol 61 pp 261ndash267 2007

[89] S Baveye E Elass D G Fernig C Blanquart J Mazurier andD Legrand ldquoHuman lactoferrin interacts with soluble CD14and inhibits expression of endothelial adhesion molecules E-selectin and ICAM-1 induced by the CD14-lipopolysaccharidecomplexrdquo Infection and Immunity vol 68 no 12 pp 6519ndash65252000

[90] C S Curran K P Demick and J M Mansfield ldquoLactoferrinactivates macrophages via TLR4-dependent and -independentsignaling pathwaysrdquo Cellular Immunology vol 242 no 1 pp23ndash30 2006

[91] P Puddu D Latorre M Carollo et al ldquoBovine lactoferrincounteracts Toll-Like receptor mediated activation signals inantigen presenting cellsrdquo PLoS ONE vol 6 no 7 Article IDe22504 2011

[92] T M Doherty and M Arditi ldquoTB or not TB That is the ques-tionmdashdoes TLR signaling hold the answerrdquoThe Journal of Clin-ical Investigation vol 114 no 12 pp 1699ndash1703 2004

[93] M A Rahman P Sobia N Gupta L Van Kaer and GDas ldquoMycobacterium tuberculosis subverts the TLR-2mdashMyD88pathway to facilitate its translocation into the cytosolrdquo PLoSONE vol 9 no 1 Article ID e86886 2014

[94] J ArtymM Zimecki andM L Kruzel ldquoEnhanced clearance ofEscherichia coli and Staphylococcus aureus in mice treated withcyclophosphamide and lactoferrinrdquo International Immunophar-macology vol 4 no 9 pp 1149ndash1157 2004

[95] G Chodaczek A Saavedra-Molina A Bacsi M L Kruzel SSur and I Boldogh ldquoIron-mediated dismutation of superoxideanion augments antigen-induced allergic inflammation effectof lactoferrinrdquo Postepy Higieny i Medycyny Doswiadczalnej vol61 pp 268ndash276 2007

[96] M Kruzel Y Harai C Chen and A Castro ldquoThe Gut a keymetabolic organ protected by lactoferrin during experimentalsystemic inflammation in micerdquo in Advances in LactoferrinResearch G Spik D Legrand J Mazurier A Pierce and J-PPerraudin Eds vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Plenum Press 1998

[97] M L Kruzel A Bacsi B Choudhury S Sur and I BoldoghldquoLactoferrin decreases pollen antigen-induced allergic airwayinflammation in a murine model of asthmardquo Immunology vol119 no 2 pp 159ndash166 2006

[98] J K Actor S-A Hwang M Olsen M Zimecki R L HunterJr and M L Kruzel ldquoLactoferrin immunomodulation of DTHresponse in micerdquo International Immunopharmacology vol 2no 4 pp 475ndash486 2002

[99] S-A Hwang M L Kruzel and J K Actor ldquoInfluence of bovinelactoferrin on expression of presentation molecules on BCG-infected bonemarrow derivedmacrophagesrdquo Biochimie vol 91no 1 pp 76ndash85 2009

[100] M Zimecki J Mazurier M Machnicki Z Wieczorek JMontreuil and G Spik ldquoImmunostimulatory activity of lacto-transferrin andmaturation of CD4minus CD8minusmurine thymocytesrdquoImmunology Letters vol 30 no 1 pp 119ndash123 1991

[101] K J Welsh S-A Hwang R L Hunter M L Kruzel and JK Actor ldquoLactoferrin modulation of mycobacterial cord factortrehalose 6-6rsquo-dimycolate induced granulomatous responserdquoTranslational Research vol 156 no 4 pp 207ndash215 2010

[102] M L Bourigault N Segueni S Rose et al ldquoRelative contri-bution of IL-1alpha IL-1beta and TNF to the host responseto Mycobacterium tuberculosis and attenuated M bovis BCGrdquoImmunity Inflammation and Disease vol 1 no 1 pp 47ndash622013

10 Mediators of Inflammation

[103] C Holscher R A Atkinson B Arendse et al ldquoA protective andagonistic function of IL-12p40 in mycobacterial infectionrdquoTheJournal of Immunology vol 167 no 12 pp 6957ndash6966 2001

[104] M Jacobs L Fick N Allie N Brown and B Ryffel ldquoEnhancedimmune response in Mycobacterium bovis bacille calmetteguerin (BCG)-infected IL-10-deficientmicerdquoClinical Chemistryand Laboratory Medicine vol 40 no 9 pp 893ndash902 2002

[105] K J Welsh S-A Hwang S Boyd M L Kruzel R L Hunterand J K Actor ldquoInfluence of oral lactoferrin onMycobacteriumtuberculosis induced immunopathologyrdquo Tuberculosis vol 91supplement 1 pp S105ndashS113 2011

[106] M L Kruzel J K Actor M Zimecki et al ldquoNovel recombinanthuman lactoferrin differential activation of oxidative stressrelated gene expressionrdquo Journal of Biotechnology vol 168 no4 pp 666ndash675 2013

[107] Y Shibata J Gabbard M Yamashita et al ldquoHeat-killedBCG induces biphasic cyclooxygenase 2+ splenic macrophageformationmdashrole of IL-10 and bone marrow precursorsrdquo Journalof Leukocyte Biology vol 80 no 3 pp 590ndash598 2006

[108] H Yamada E Kuroda S Matsumoto T Matsumoto TYamada and U Yamashita ldquoProstaglandin E

2down-regulates

viable Bacille Calmette-Guerin-induced macrophage cytotoxi-city against murine bladder cancer cell MBT-2 in vitrordquoClinicaland Experimental Immunology vol 128 no 1 pp 52ndash58 2002

[109] S Hwang M L Kruzel and J K Actor ldquoEffects of CHO-expressed recombinant lactoferrins on mouse dendritic cellpresentation and functionrdquo Innate Immunity vol 21 no 5 pp553ndash561 2015

[110] K M OShea S-A Hwang and J K Actor ldquoImmune activity ofbcg infected mouse macrophages treated with a novel recom-binant mouse lactoferrinrdquo Annals of Clinical Lab Science vol45 no 5 pp 403ndash410 2015

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Mediators of Inflammation 9

[74] M L Kruzel and M Zimecki ldquoLactoferrin and immunologicdissonance clinical implicationsrdquo Archivum Immunologiae etTherapiae Experimentalis vol 50 no 6 pp 399ndash410 2002

[75] W I Weis M E Taylor and K Drickamer ldquoThe C-type lectinsuperfamily in the immune systemrdquo Immunological Reviewsvol 163 pp 19ndash34 1998

[76] W I Weis K Drickamer and W A Hendrickson ldquoStructureof a C-type mannose-binding protein complexed with an oli-gosacchariderdquo Nature vol 360 no 6400 pp 127ndash134 1992

[77] T Zagulski Z Jarzabek A Zagulska M Jaszczak I EKochanowska and M Zimecki ldquoLactoferrin stimulates killingand clearance of bacteria but does not prevent mortality of dia-betic micerdquo Archivum Immunologiae et Therapiae Experimen-talis vol 49 no 6 pp 431ndash438 2001

[78] M Kruzel Y Harai C Chen and A Castro ldquoRole of lactoferrinin development of systemic inflammatory response syndrome(SIRS)rdquo in Proceedings of the 2nd International ConferenceProgress in Intensive Care Medicine pp 11ndash12 Wroclaw Poland1998

[79] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoThe guta key metabolic organ protected by lactoferrin during experi-mental systemic inflammation in micerdquo in Advances in Lacto-ferrin Research vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Springer New York NY USA 1998

[80] MKruzel T Zagulski andMZimecki ldquoLactoferrin and insult-induced metabolic imbalance in humans and other animalsrdquoin Lactoferrin Structure Function and Applications Proceedingsof the 4th International Conference on Lactoferrin StructureFunction and Applications held in Sapporo Japan 18ndash22 May1999 pp 301ndash311 ICS Elsevier New York NY USA 2000

[81] M L Kruzel Y Harari C-Y Chen and G A Castro ldquoLactofer-rin protects gut mucosal integrity during endotoxemia inducedby lipopolysaccharide in micerdquo Inflammation vol 24 no 1 pp33ndash44 2000

[82] E Zaczynska M Kocięba E Sliwinska and M ZimeckildquoBovine lactoferrin enhances proliferation of human peripheralblood lymphocytes and induces cytokine production in wholeblood culturesrdquo Advances in Clinical and Experimental Medi-cine vol 23 pp 871ndash876 2014

[83] Z Wei T Nishimura and S Yoshida ldquoCharacterization ofglycans in a lactoferrin isoform lactoferrin-ardquo Journal of DairyScience vol 84 no 12 pp 2584ndash2590 2001

[84] B-K Choi J K Actor S Rios et al ldquoRecombinant humanlactoferrin expressed in glycoengineered Pichia pastoris effectof terminal N-acetylneuraminic acid on in vitro secondaryhumoral immune responserdquoGlycoconjugate Journal vol 25 no6 pp 581ndash593 2008

[85] B Samyn-Petit J-PWajdaDubos F Chirat et al ldquoComparativeanalysis of the site-specific N-glycosylation of human lactofer-rin produced in maize and tobacco plantsrdquo European Journal ofBiochemistry vol 270 no 15 pp 3235ndash3242 2003

[86] M Zimecki J Artym M Kocięba M Duk and M L KruzelldquoThe effect of carbohydrate moiety structure on the immu-noregulatory activity of lactoferrin in vitrordquo Cellular ampMolecu-lar Biology Letters vol 19 no 2 pp 284ndash296 2014

[87] J K Actor S-A Hwang and M L Kruzel ldquoLactoferrin as anatural immune modulatorrdquo Current Pharmaceutical Designvol 15 no 17 pp 1956ndash1973 2009

[88] M L Kruzel J K Actor I Boldogh and M ZimeckildquoLactoferrin in health and diseaserdquo Postepy Higieny i MedycynyDoswiadczalnej vol 61 pp 261ndash267 2007

[89] S Baveye E Elass D G Fernig C Blanquart J Mazurier andD Legrand ldquoHuman lactoferrin interacts with soluble CD14and inhibits expression of endothelial adhesion molecules E-selectin and ICAM-1 induced by the CD14-lipopolysaccharidecomplexrdquo Infection and Immunity vol 68 no 12 pp 6519ndash65252000

[90] C S Curran K P Demick and J M Mansfield ldquoLactoferrinactivates macrophages via TLR4-dependent and -independentsignaling pathwaysrdquo Cellular Immunology vol 242 no 1 pp23ndash30 2006

[91] P Puddu D Latorre M Carollo et al ldquoBovine lactoferrincounteracts Toll-Like receptor mediated activation signals inantigen presenting cellsrdquo PLoS ONE vol 6 no 7 Article IDe22504 2011

[92] T M Doherty and M Arditi ldquoTB or not TB That is the ques-tionmdashdoes TLR signaling hold the answerrdquoThe Journal of Clin-ical Investigation vol 114 no 12 pp 1699ndash1703 2004

[93] M A Rahman P Sobia N Gupta L Van Kaer and GDas ldquoMycobacterium tuberculosis subverts the TLR-2mdashMyD88pathway to facilitate its translocation into the cytosolrdquo PLoSONE vol 9 no 1 Article ID e86886 2014

[94] J ArtymM Zimecki andM L Kruzel ldquoEnhanced clearance ofEscherichia coli and Staphylococcus aureus in mice treated withcyclophosphamide and lactoferrinrdquo International Immunophar-macology vol 4 no 9 pp 1149ndash1157 2004

[95] G Chodaczek A Saavedra-Molina A Bacsi M L Kruzel SSur and I Boldogh ldquoIron-mediated dismutation of superoxideanion augments antigen-induced allergic inflammation effectof lactoferrinrdquo Postepy Higieny i Medycyny Doswiadczalnej vol61 pp 268ndash276 2007

[96] M Kruzel Y Harai C Chen and A Castro ldquoThe Gut a keymetabolic organ protected by lactoferrin during experimentalsystemic inflammation in micerdquo in Advances in LactoferrinResearch G Spik D Legrand J Mazurier A Pierce and J-PPerraudin Eds vol 443 of Advances in Experimental Medicineand Biology pp 167ndash173 Plenum Press 1998

[97] M L Kruzel A Bacsi B Choudhury S Sur and I BoldoghldquoLactoferrin decreases pollen antigen-induced allergic airwayinflammation in a murine model of asthmardquo Immunology vol119 no 2 pp 159ndash166 2006

[98] J K Actor S-A Hwang M Olsen M Zimecki R L HunterJr and M L Kruzel ldquoLactoferrin immunomodulation of DTHresponse in micerdquo International Immunopharmacology vol 2no 4 pp 475ndash486 2002

[99] S-A Hwang M L Kruzel and J K Actor ldquoInfluence of bovinelactoferrin on expression of presentation molecules on BCG-infected bonemarrow derivedmacrophagesrdquo Biochimie vol 91no 1 pp 76ndash85 2009

[100] M Zimecki J Mazurier M Machnicki Z Wieczorek JMontreuil and G Spik ldquoImmunostimulatory activity of lacto-transferrin andmaturation of CD4minus CD8minusmurine thymocytesrdquoImmunology Letters vol 30 no 1 pp 119ndash123 1991

[101] K J Welsh S-A Hwang R L Hunter M L Kruzel and JK Actor ldquoLactoferrin modulation of mycobacterial cord factortrehalose 6-6rsquo-dimycolate induced granulomatous responserdquoTranslational Research vol 156 no 4 pp 207ndash215 2010

[102] M L Bourigault N Segueni S Rose et al ldquoRelative contri-bution of IL-1alpha IL-1beta and TNF to the host responseto Mycobacterium tuberculosis and attenuated M bovis BCGrdquoImmunity Inflammation and Disease vol 1 no 1 pp 47ndash622013

10 Mediators of Inflammation

[103] C Holscher R A Atkinson B Arendse et al ldquoA protective andagonistic function of IL-12p40 in mycobacterial infectionrdquoTheJournal of Immunology vol 167 no 12 pp 6957ndash6966 2001

[104] M Jacobs L Fick N Allie N Brown and B Ryffel ldquoEnhancedimmune response in Mycobacterium bovis bacille calmetteguerin (BCG)-infected IL-10-deficientmicerdquoClinical Chemistryand Laboratory Medicine vol 40 no 9 pp 893ndash902 2002

[105] K J Welsh S-A Hwang S Boyd M L Kruzel R L Hunterand J K Actor ldquoInfluence of oral lactoferrin onMycobacteriumtuberculosis induced immunopathologyrdquo Tuberculosis vol 91supplement 1 pp S105ndashS113 2011

[106] M L Kruzel J K Actor M Zimecki et al ldquoNovel recombinanthuman lactoferrin differential activation of oxidative stressrelated gene expressionrdquo Journal of Biotechnology vol 168 no4 pp 666ndash675 2013

[107] Y Shibata J Gabbard M Yamashita et al ldquoHeat-killedBCG induces biphasic cyclooxygenase 2+ splenic macrophageformationmdashrole of IL-10 and bone marrow precursorsrdquo Journalof Leukocyte Biology vol 80 no 3 pp 590ndash598 2006

[108] H Yamada E Kuroda S Matsumoto T Matsumoto TYamada and U Yamashita ldquoProstaglandin E

2down-regulates

viable Bacille Calmette-Guerin-induced macrophage cytotoxi-city against murine bladder cancer cell MBT-2 in vitrordquoClinicaland Experimental Immunology vol 128 no 1 pp 52ndash58 2002

[109] S Hwang M L Kruzel and J K Actor ldquoEffects of CHO-expressed recombinant lactoferrins on mouse dendritic cellpresentation and functionrdquo Innate Immunity vol 21 no 5 pp553ndash561 2015

[110] K M OShea S-A Hwang and J K Actor ldquoImmune activity ofbcg infected mouse macrophages treated with a novel recom-binant mouse lactoferrinrdquo Annals of Clinical Lab Science vol45 no 5 pp 403ndash410 2015

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

10 Mediators of Inflammation

[103] C Holscher R A Atkinson B Arendse et al ldquoA protective andagonistic function of IL-12p40 in mycobacterial infectionrdquoTheJournal of Immunology vol 167 no 12 pp 6957ndash6966 2001

[104] M Jacobs L Fick N Allie N Brown and B Ryffel ldquoEnhancedimmune response in Mycobacterium bovis bacille calmetteguerin (BCG)-infected IL-10-deficientmicerdquoClinical Chemistryand Laboratory Medicine vol 40 no 9 pp 893ndash902 2002

[105] K J Welsh S-A Hwang S Boyd M L Kruzel R L Hunterand J K Actor ldquoInfluence of oral lactoferrin onMycobacteriumtuberculosis induced immunopathologyrdquo Tuberculosis vol 91supplement 1 pp S105ndashS113 2011

[106] M L Kruzel J K Actor M Zimecki et al ldquoNovel recombinanthuman lactoferrin differential activation of oxidative stressrelated gene expressionrdquo Journal of Biotechnology vol 168 no4 pp 666ndash675 2013

[107] Y Shibata J Gabbard M Yamashita et al ldquoHeat-killedBCG induces biphasic cyclooxygenase 2+ splenic macrophageformationmdashrole of IL-10 and bone marrow precursorsrdquo Journalof Leukocyte Biology vol 80 no 3 pp 590ndash598 2006

[108] H Yamada E Kuroda S Matsumoto T Matsumoto TYamada and U Yamashita ldquoProstaglandin E

2down-regulates

viable Bacille Calmette-Guerin-induced macrophage cytotoxi-city against murine bladder cancer cell MBT-2 in vitrordquoClinicaland Experimental Immunology vol 128 no 1 pp 52ndash58 2002

[109] S Hwang M L Kruzel and J K Actor ldquoEffects of CHO-expressed recombinant lactoferrins on mouse dendritic cellpresentation and functionrdquo Innate Immunity vol 21 no 5 pp553ndash561 2015

[110] K M OShea S-A Hwang and J K Actor ldquoImmune activity ofbcg infected mouse macrophages treated with a novel recom-binant mouse lactoferrinrdquo Annals of Clinical Lab Science vol45 no 5 pp 403ndash410 2015

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom