Laboratory for Immunohistochemistry and Immunopathology (LIIPAT), Department of Pathology,

Oslo University Hospital Rikshospitalet, Norway

Per Brandtzaegper.brandtzaeg@medisin.uio.no

Mucosal immunization:relevance to protection against

tuberculosis

The mucosae are an enormous battlefieldMucosal effector sites provide secretory IgA (SIgA) antibodies

Section through skin

Airways and oral cavity

Gastrointestinal mucosa

Hornified layerEpithelial cells

Mucus and cilia

GlandsPlasma cells

Surface epithelium

Glands (crypts)

Plasma cells

Epithelial cells

IgAIgGNormal human colon

Plasma cellsH&E

IgA

80% of all plasma cells arelocated in gut mucosa

An adult exports 3 g of SIgA to the gut lumen per day

At the border of hell!

Local formation and export of mucosal immunoglobulins

Dimeric IgA (pIgA)

J chain

Mucus

Gland

IgA+J

SIgM

SIgA

IgM+J

LumenLamina propria

IgG

pIgR(mSC)

PentamericIgM IgG(±J)

Free SC

IgA+J

Plasma cells

IgA

IgA-coated bacteria

Brandtzaeg Nature & J. Immunol. 1974; Brandtzaeg & Prydz Nature 1984

Ag

‘The IgA pump’

Containment

Control, not clearance, provides

mutualism

Coating of bacteria with SIgA is an important immune exclusion

mechanism

IgA-producing cellsModified from: Natalia Shulzhenko … Polly Matzinger, Nature Med 2011; 17: 1585-93

(PRR)

pIgR

•SIgA-coated bacteria are less invasive

•Less shedding of SIgA-coated bacteria

•Less horizontal spread of SIgA-coated pathogens (provides herd protection)

(SIgA)

Bronchioles(< 1 mm):Serum-derivedIgG

Trachea andbronchi:Secretory IgA

Defence of lung parenchyma relies largely on serum-derived IgG and cell-mediated mechanisms

The lung is not a gut that breathes

Bronchioles(< 1 mm):Serum-derivedIgG

Trachea andbronchi:Secretory IgA

Defence of lung parenchyma relies largely on serum-derived IgG and cell-mediated mechanisms

The lung is not a gut that breathes

Gereke et al., Respir. Res. 2007; 8: 47

Type IIpneumocyte

ALVEOLUS

Marcus Gereke (Braunschweig, Germany):Alveolar type II epithelial cells: pneumocytes with regulatory properties, also anti-inflammatory

Homeostatic function ofmucosal vaccines

• The goal of mucosal vaccines is to stop the pathogen at the portal of entry

• This can best be achieved by induction of secretory IgA (SIgA) antibodies

• Protection against invasive mucosal pathogens requires, in addition, systemic immunity (IgG antibodies and cytotoxic T cells)

Small intestinalmucosa

Large Intestinal mucosa

Peyer'spatches(GALT)

Uro-genitaltract

Mammaryglands

Lacrimal,nasal, &salivary glands

Bronchial glands

Regionalization in the integrated mucosal immune systemTonsils &adenoids(NALT)

BALT

Dichotomy in the integrated mucosal immune system

Tonsils(NALT)

BALT

Peyer’spatches(GALT)

Isolated lymphoid follicles (GALT)

Also systemic

Also systemic

Human palatine tonsil

Human NALT anlagen: prenatal (19 wks).Nasal ILFs in 40% < 2 yrs (inducible?).Rodent NALT: postnatal organogenesis.

Tonsils and adenoids are well designed for antigen trappingInductive sites

BALT

Peyer'spatches,appendixand solitary intestinal lymphoid follicles (GALT)

Tonsils &adenoids(NALT)

GC

Crypt

Scanning electron-microscopy (Owen, 1988)

Reticular cryptepithelium(cytokeratin)

Reticular crypt epithelium (cytokeratin)

BALTHuman BALT in 40%-100% at young age (2-15 yrs); rare in adult normal lungs (>20 yrs), but inducible (also rodent BALT is inducible)

Inductive sites

BALT

Tonsils &adenoids(NALT)

Human NALT anlagen prenatal (19 wks).Nasal ILFs in 40% < 2 yrs (inducible?).Rodent NALT: postnatal organogenesis

Human BALT in 40%-100% at young age (2-15 yrs); rare in adult normal lungs (>20 yrs), but inducible (also rodent BALT is inducible)

Human bronchus-associtaed lymphoid tissue (BALT)

Submucosal glands

Adult lung with chronic infection

Inductive mucosa-associated lymphoid tissue (MALT) in human airways

HLA-DR HLA-DR

DC

DC

CD3(T)FoxP3(Treg)CD3(T)CD20(B)

Adenoids

Middle turbinate

Oral cavity

Tounge

Epiglottis

Inferior turbinate

Olfactory region

Tubal tonsil

Palatine tonsil

Lingual tonsil

Skin

Waldeyer’s ringOrganized lymphoid tissue with M cells

Nasal anatomy and location of regional lymphoid tissue

Olfactory region

Ciliated mucosa

Airway mucosais extremely rich in dendritic cells

150-200 cm2

Jahnsen FL, Gran E, Haye R, Brandtzaeg P. Am J Respir Cell Mol Biol, 2004; 30:31-37

HLA-DRRFD-7

Human nasal mucosa

Mesentericlymph nodes

Thoracicduct Cervical

lymph nodes

Right lymphaticduct

Regional lymph drainage through mesenteric and cervical lymph nodes

Mesentericlymph nodes

Thoracicduct Cervical

lymph nodes

Right lymphaticduct

Regional lymph drainage through mesenteric and cervical lymph nodes

Sublingual vaccination

Immune compart-ments of the lung

From: Holt et al. Nature Rev. Immunol. 2008; 8:142-52

CD4+ T cells only become activated onceM. tuberculosis spreads from the lungs tothe lymph nodes and starts to produce antigen

Wolf et al.J. Exp. Med.2008 205: 105-15

Airway luminal T cells: a newcomer on the stage of TB vaccination strategies

Jeyanathan M, Heriazon A, Xing Z. Airway luminal T cells: a newcomer on the stage of TB vaccination strategies. Trends Immunol. 2010; 31: 247-52

pIgR/SC knockout mice lack epithelial IgA transportIgA IgG+/+ –/–

Johansen et al. J. Exp. Med. 1999; 190: 915-21

Mice i.n. vaccinated with PstS-1 antigen (plus CT) and then i.n. infected

Naïve mice infected i.n. with 106 CFUs of BCG

Effect of mucophilic SIgA and the ciliary conveyer band?

                                          Proceedings

Passive administration of purified secretory IgA from human colostrum induces protection against Mycobacterium tuberculosis in a murine model of progressive pulmonary infection

Nadine Alvarez1

, Oscar Otero1

, Frank Camacho1

, Reinier Borrero1

, Yanely Tirado1

, Alina Puig1

, Alicia Aguilar1

, Cesar Rivas2

, Axel Cervantes2

, Gustavo Falero-Díaz1

, Armando Cádiz3

, María E Sarmiento1, Mohd Nor Norazmi4,5

, Rogelio Hernández-Pando2 and Armando Acosta1*

•

* Corresponding author: Armando Acosta aracosta2001@yahoo.com Author Affiliations

1 Department of Molecular Biology. Finlay Institute. Center of Research – Producction of Vaccines. Ave. 27 No. 19805, La Lisa. Ciudad de la Habana, Cuba. AP. 16017, CP 11600 2 Experimental Pathology Section, Department of Pathology, National Institute of Medical Sciences and Nutrition “Salvador Zubiràn”, D.F. Mexico. CP 14 000 3 Enterprise of Production of Serum and Hemoderivates “Adalberto Pesant González”. Ave 51 No.33 235 km 19 medio ½. Arroyo Arenas, La Lisa. Ciudad de la Habana, Cuba. CP 13400 4 School of Health Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Malaysia 5 Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 16150 Kubang Kerian, Malaysia For all author emails, please log on. BMC Immunology 2013, 14(Suppl 1):S3 doi:10.1186/1471-2172-14-S1-S3

Authors from Cuba, Mexico and Malaysia

Determination of bacterial load (A) and pneumonic area (B) in lungs of mice which were untreated (NT) and those treated with human secretory IgA (hsIgA), after challenge with M. tuberculosis H37Rv by intratracheal route 2 hrs after inoculation.

Another group received M. tuberculosis preincubated with hsIgA (preinc). Well-organized granuloma of preincubated group 2 months after challenge with M. tuberculosis, visualized by H&E staining (25x) (C).

• Such vaccine administration elicits both regional mucosal and systemic immunity

• Future strategy: prime-boost approach, e.g. BCG (prime) followed by mucosal boost, or vice versa

Acknowledgements

Laboratory for Immunohistochemistry and Immunopathology (LIIPAT) is part of Centre for Vaccinology and

Immunotherapy (CEVI, 2001) and Centre of Excellence for Immune Regulation (CIR, 2007), funded by the Research Council of Norway, University of Oslo and Rikshospitalet

University Hospital