Pulmonary mucosal BCG vaccination shows protection of...

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PulmonarymucosalBCGvaccinationshowsprotectionofinfectioninanovelrepeatedultra-low

dosechallengemodelinrhesusmacaques.FrankVerreck

5thGlobalForumonTBVaccines[OA-04]21February2018,NewDelhi

Biomedical Primate Research Centre Committed to health researchand alternatives

UsingMacaqueSpeciesinTBVaccineResearch

tosupportpreclinicalvaccinedevelopmentbyefficacytesting

toinvestigatemechanismsofdiseaseandprotectiveinfection

exploratory rsrch

applied rsrch

Macacamulattarhesus

Macacafasciculariscynomolgus

vaccine R&D

biomarker R&D

translation

back-translation& verification

• naturallysusceptibletoMtb• greatfacevalidityofTBinmanyaspects

Biomedical Primate Research Centre Committed to health researchand alternatives

Today'sPresentation

• onoptimizingmodelconditionsforvaccineefficacyevaluationinrhesusmacaquesultimatelydemonstratingthefeasibilityofRepeatedUltra-LowDose(RULD)infectionasusefulmodellingcondition

• oninvestigatingmechanismsofdiseaseandofprotectiveimmunity,towardscorrelates/biomarkersfindinglocalpolyfunctionalIL17+CD4cellsandspecificimmunoglobulinsasdistinctivemarkersofprotection

Biomedical Primate Research Centre Committed to health researchand alternatives

Standardising NHPTBStudiesforVaccineResearch

hostRhesus(Macacamulatta)M.tuberculosischallengestrainErdmanK01(harmonisationstrain,fromBEIResources,USA)byendobronchialinstillationBCGvaccinestandardhumandoseofstrainMoscow(a.k.a.Sophia),whileinearlierstudiesusingDanish1331

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SingleHighDose(500CFU)MtbChallenge

time-to-humane-endpoint (survival)highdose

Modusofreadout

ModusofInfection

12monthsfollow-up,groupsizeN=12Verreck etal(unpublished)

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SingleHighDose(500CFU)MtbChallenge

time-to-humane-endpoint (survival)highdose

Modusofreadout

ModusofInfection

advanced diseasehighdose

✓

12weeksfollow-up,groupsizeN=6Verreck etal(PLoSOne)2009

Biomedical Primate Research Centre Committed to health researchand alternatives

IsHighDoseChallengeinRhesusTooStringentforTBVaccineEvaluation?

time-to-humane-endpoint (survival)highdose

Modusofreadout

ModusofInfection

advanced diseasehighdose

✓• 100%infection takeinnv.ctrls• rapid onset toprogressiveTB(≤12wks)• powerto demonstrate reductionofTB

(relative to nv.ctrls) with group size N=6only

BUT• unable to demonstrate

statistically significantimprovement overBCG;findingpositiveindicatorsonly

✓

Biomedical Primate Research Centre Committed to health researchand alternatives

PulmonaryBCGprotectswherestandardBCGfails

time-to-humane-endpoint (survival)highdose

Modusofreadout

ModusofInfection

advanced diseasehighdose

✓

✓

cohortA

Corroboratingandextendingbeyondearlierstudiesshowingthesuperiorityofi.v.>pulm>i.dBCGadministrationinNHPBarclayetal(1970)AmJRev;

Barclayetal(1973)IAIVerreck etal(2017)

Tuberculosis

cohortB

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Working towards "low"dose infection modelling

time-to-humane-endpoint (survival)highdose

advanced diseasehighdose

milddiseaselowdose

infection(and/ormilddisease)

repeatedlowdose

infection(a/omilddisease)

naturaltransmission

Modusofreadout

ModusofInfection

✓

✓

✓

?

12monthsfollow-up,groupsizeN=12

12weeksfollow-up,groupsizeN=6

10to25CFUNHPTBResearchCommunityofCTVD

Biomedical Primate Research Centre Committed to health researchand alternatives

FindingthelowestamenabledoseforRLDinfectionacomparativeinfectiondose-escalationstudyinrhesusversuscynomolgus

challenge(0.5(CFU(

immune((monitoring(

+ve(

yes$

no$

challenge(2.5(CFU(

immune((monitoring(

+ve(

yes$

no$

challenge(12.5(CFU(

immune((monitoring(

+ve(

yes$

no$

challenge(62.5(CFU(

immune((monitoring(

+ve(

yes$

endpoint(evalua<on(@(wk(6+(

endpoint(evalua<on(@(wk(12+(

if$N≥5,$then$for$N=½n$

endpoint(evalua<on(@(wk(6+(

endpoint(evalua<on(@(wk(12+(

if$N≥5,$then$for$N=½n$

endpoint(evalua<on(@(wk(6+(

endpoint(evalua<on(@(wk(12+(

if$N≥5,$then$for$N=½n$

endpoint(evalua<on(@(wk(6+(

endpoint(evalua<on(@(wk(12+(

if$N≥5,$then$for$N=½n$

0 4 8 12

0

25

50

75

100

Study Weeks (post-primary infection)

%

Time to IGRA conversion

CynoRhesus

0.2 CFU 1.3 CFU 7 CFU

K.Dijkman etal.(inpreparation)VisitposterPD-10,ondiseasesusceptibilityandimmuneprofilesofrhesusversuscyno

KarinDijkman,Ph.D student

Biomedical Primate Research Centre Committed to health researchand alternatives

Repeated Ultra-LowDose (RULD)Infectious Challengean alternative study designinrhesus (N=8pergroup)toward readout ofPoI

K.Dijkman etal.(inpreparation)

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IGRAresponsedynamics overtimeby specific IFNg ELISPOT

PPDstimulated ESAT6-CFP10stimulated

K.Dijkman etal.(inpreparation)

Biomedical Primate Research Centre Committed to health researchand alternatives

Mucosal BCGshowssignificantly delayedIGRAconversion,aPoIsignal

K.Dijkman etal.(inpreparation)

Biomedical Primate Research Centre Committed to health researchand alternatives

Mucosal BCGsignificantly reduces pathology

K.Dijkman etal.(inpreparation)

Biomedical Primate Research Centre Committed to health researchand alternatives

Mucosal BCGsignificantly reduces bacterialload

K.Dijkman etal.(inpreparation)

pathology

bacteriology

Biomedical Primate Research Centre Committed to health researchand alternatives

CD4Tcells inBALby multi-labelflow-cytometry,8weekspost-vaccination

mucosal BCGinduces highest cyk levelsandan exclusive IL17signal

IFNg TNFa IL-2 IL-17A

NVIFNg

BCGIFNg

MUCIFNg

0

2

4

6

20

40

60

80

% c

yto

kin

e p

ost

ive

0.000155

0.0006220.00109

NVTNFa

BCGTNFa

MUCTNFa

0

2

4

6

20

40

60

80%

cyt

oki

ne

po

stiv

e0.000155

0.0006220.00171

NVIL-2

BCGIL-2

MUCIL-2

0

2

4

6

20

40

60

80

% c

yto

kin

e p

ost

ive

0.000155

0.0003110.000622

NVIL-17A

BCGIL-17A

MUCIL-17A

0

2

4

6

20

40

60

80

% c

yto

kin

e p

ost

ive

0.000155

0.0001550.0595

*** *** *** ********* ****** ****** ***

K.Dijkman etal.(inpreparation)

Biomedical Primate Research Centre Committed to health researchand alternatives

mucosal BCGexcl.induces alocal poly-functionalTh17response

K.Dijkman etal.(inpreparation)

wk8post-vaccination

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mucosal BCGinducesspecific localIgGandIgA

K.Dijkman etal.(inpreparation)

BALflu

id

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SummarizingConclusionsRULDchallengeisafeasiblestrategyforenhancedvaccineefficacyreadoutinrhesustowardPoIandPoDsignals

showingmoresubtleTBinfectionanddiseasephenotypes

PulmonarymucosalBCGvaccinationinNHP(again)issuperioroverstandardintradermal BCGadministration

bydelayedIGRAconversionandsignificantreduction ofpathologyandbacterialload(noCFUdetectablein2outof8)

Local,Ag-specific,multi-functional IL17+CD4+Tcellsand IgG&IgA appearascorrelatesofprotectiveimmunity

LocalmucosalvaccinationperseandperhapspulmonaryBCGadministrationinparticular,providesagreatperspectiveforimprovedBCGvaccinationorBCGre-vaccinationtofightTB

Biomedical Primate Research Centre Committed to health researchand alternatives

AcknowledgementsBPRC,TBResearch&Immunology

KarinDijkman

RichardVervenneClaudiaSombroekSamHofmanCharelBoot

M.A.KhayumKristaHaanstraMichelVierboom

BPRC,AnimalScience DepartmentPathology TeamClincal LabAnimalCARE TeamVetCARE Team

LUMC,LeidenTomOttenhoffc.s.