Biomedical Primate Research Centre Committed to health researchand alternatives
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
Biomedical Primate Research Centre Committed to health researchand alternatives
SingleHighDose(500CFU)MtbChallenge
time-to-humane-endpoint (survival)highdose
Modusofreadout
ModusofInfection
12monthsfollow-up,groupsizeN=12Verreck etal(unpublished)
Biomedical Primate Research Centre Committed to health researchand alternatives
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
Biomedical Primate Research Centre Committed to health researchand alternatives
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)
Biomedical Primate Research Centre Committed to health researchand alternatives
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
Biomedical Primate Research Centre Committed to health researchand alternatives
mucosal BCGinducesspecific localIgGandIgA
K.Dijkman etal.(inpreparation)
BALflu
id
Biomedical Primate Research Centre Committed to health researchand alternatives
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.
