zika virus infection in cynomolgus macaques...zika natural history study in cynomolgus macaques...
TRANSCRIPT
Zika Virus Infection in Cynomolgus Macaques Fusataka Koide Program Leader, Emerging Pathogens Southern Research
SouthernResearch
• Mosquito-bornevirusbelongingtotheflavivirusgenusoftheFlaviviridaefamily.
• Dengueviruses,WestNileVirus,St.LouisEncephaliBsvirus
• SmallviruseswithanRNAgenomethatishighlymutable
• TransmiFedbetweenvertebratehosts(humansandprimates)byAedesspeciesmosquitoes
Background
SouthernResearch
GlobalSpreadofZikaVirus
SouthernResearch
TransmissionofZikaVirus
SouthernResearch
ZikaInfecBoninHumans
Guillain–Barrésyndrome- Autoimmunedisorder- Peripheralnervoussystemimpairment
hFps://en.wikipedia.org/wiki/Zika_virus_outbreak_(2015present)#Guillain.E2.80.93Barr.C3.A9_syndrome
NoFDAApprovedVaccinesorTherapeuBcsAvailableforZika
SouthernResearch
ZikaAnimalModelsØ A129MouseModel(lethal)Ø AG129MouseModel(lethal)Ø WildtypeMouse(non-lethal,viremiafollowingIVinfecBon)Ø WildtypeMouse(lethal,typeIIFNblockade)Ø IndianRhesusMacaques(Viremia,Immunology,Microcephaly)
SouthernResearch
ü Humanchallengestudymynotbefeasibleü AnimalmodelrecapitulaBnghumaninfecBonisneeded
WhyMauriBanCynomolgusMacaque(MCM)forZika?
Ø MCMproducedrobust immunogenicitydata inthepastDengueLATDV*studies
Ø Sustainable viremia following infecBon with Dengue,YellowFeverorWestNileviruses
Ø HavelowFlavivirussero-prevalence(<0.6%)*LiveaFenuatedtetravalentdenguevaccine
SouthernResearch
WhyMauriBanCynomolgusMacaque(MCM)forZika?
Ø SmalleranddocilethanIndianrhesusmacaques(IRM)
Ø ReadilyavailablecomparedtoIRM
Ø Lesscostly
Ø ScienBficjusBficaBonsandlogisBcaladvantage
SouthernResearch
ZikaNaturalHistoryStudyinCynomolgusMacaques(N=6)
Group! N Sex! Virus! Challenge strain!(Day 0)! Virus lineage!
Challenge Volume
(SC)!Challenge Dose!
1! 1! M! ZIKV! PRVABC59! Asian 0.5 mL ! 5.0 x105 PFU!2! 1! F! ZIKV! PRVABC59! Asian 0.5 mL ! 1.0 x104 PFU!3! 1! M! ZIKV! FSS13025! Asian 0.5 mL ! 5.0 x105 PFU!4! 1! F! ZIKV! FSS13025! Asian 0.5 mL ! 1.0 x104 PFU!5! 1! M! ZIKV! IBH 30656! African 0.5 mL ! 5.0 x105 PFU!6! 1! F! ZIKV! IBH 30656! African 0.5 mL ! 1.0 x104 PFU!
MainObjecBves:ü Testabilitytosustainviremiaahersubcutaneouschallenge
ü DetecBonofneutralizinganBbody(Nab)postchallenge
SouthernResearch
Day-20123456810143060
ZikaChallenge
Serum,UrineandSalivaCollecBon
Orchiectomy
SerumCollecBon
EuthanasiaandTissueHarvest
ZikaNaturalHistoryStudyinCynomolgusMacaques(N=6)
Serum,UrineandSalivacollecBon SerumcollecBon
Orchiectomy
SouthernResearch
ZikaSerumViralLoadinCynomolgusmacaques
Gen
omecopies/m
L
DaysPostInfec;on
1.E+0
1.E+1
1.E+2
1.E+3
1.E+4
1.E+5
1.E+6
PRVABC59
PRVABC59
FSS13025
FSS13025
IBH30656
IBH30656
LLOQ
12346810143060
InfecBonwithPRVABCandFSSbutnotwithIbHSouthernResearch
ViralLoadintheTestesofZIKVinfectedCynomolgusmacaques
*
*
D4D8D4D8D4D8PRVABC59FSS13025IBH30656
Gen
omecopies/g;ssue
*p-value<0.05
Againconsistentwiththeserumanalysis,noviralloadwasdetectedinthetestesofanimalchallengedwiththeAfricanstrainIBH30656.SouthernResearch
Day Urine OralSwabs PRVABC59 FSS13025 IBH30656 PRVABC59 FSS13025 IBH30656 #5262M #5258F #5260M #5259
F#5261M
#5257F #5262M
#5258F
#5260M
#5259
#5261M
#5257F
1 0 0 0 N/C 0 N/C 0 0 0 1.6 N/T N/T2 265 0 N/C N/C 43 N/C 105 0 0 0 N/T N/T3 0 0 19 N/C N/C N/C 198 0 52 0 N/T N/T4 0 N/C N/C N/C 0 N/C 169 146 0 1 N/T N/T6 0 29 0 N/C 0 N/C 0 0 6.6 2 N/T N/T8 0 0 N/C N/C 0 N/C 138 142 0 81 N/T N/T10 0 87 0 N/C 200 N/C 244 0 12 0 N/T N/T14 0 114 N/C N/C 0 N/C 109 359 0 0 N/T N/T
ViralSheddingfromBiologicalFluids
ViralSheddingUrinewascollecteddirectlyfromanimals(asopposedtocagepans).Detectedsporadicverylow<300copies/mLinurineand<400copies/mLinsaliva.Asexpected,animalsfromtheAfricanlineageIBH30656challenge,exhibitedextremelylowlevelsofvirusshedding.
SouthernResearch
ZIKVInfecBoninCynomolgusMacaquesViralLoadbyqRT-PCR(urineandsaliva)
0
50
100
150
200
250
300
0 2 4 6 8 10 12 14 16
Copies/m
LUrin
eorSaliva
DaysPostInoculaBon
ChallengeStrainZKVPuertoRicoPRVABC59
#5262Urine#5262Saliva
ProlongedZIKVsheddingobservedinthesalivaofinfectedMCM
SouthernResearch
NeutralizingAnBbodyResponse
1
10
100
1000
10000
5262 5258 5260 5259
PRNT50Titer
AnimalID
PRNT50TitertoPRVABC59
Day14 Day30 Day60
SouthernResearch
NeutralizingAnBbodyResponsetoPRVABC59
Animal ID Challenge
Strain Dose
(PFU) PRNT50 Titer D-2 D14 D30 D60
5262 M PRVABC59 5.0 x 105 < 10 7920 2380 1071 5258 F PRVABC59 1.0 x 104 < 10 2438 875 740 5260 M FSS13025 5.0 x 105 < 10 1053 797 528 5259 F FSS13025 1.0 x 104 < 10 3131 2019 666 5261 M IBH 30656 5.0 x 105 < 10 < 10 < 10 < 10 5257 F IBH 30656 1.0 x 104 < 10 < 10 < 10 < 10
SouthernResearch
ü ThemonkeysinoculatedwiththePRVandFSSachievedhighNabBtertoPRVbyday14.
ü TheIBHchallengedmonkeys(5261and5257)thatexhibitedverylowviralloadthroughoutstudyperiodalsofailedtoproducecrossreacBveNabtoPRVABC59.
Developmentof96-wellFRNTassaytoacceleratedevelopmentofZikacountermeasures
Cellsfixedandstainedonly24hoursaherinfecBoncombinedwithautomatedcounBngoffociincreasesturnaroundBmecomparedtostandardPRNTassaySouthernResearch
CytokineandChemokineAnalysisintheSerum
0 1 2 3 4 6 8 1 0 1 4 3 0 6 00
5 0
1 0 0
1 5 0
2 0 04 0 0
5 0 0
6 0 0
7 0 0
8 0 0
T im e P o s t- In fe c tio n (D a y s )
IL-1
Ra
(p
g/m
l)
P R (M )
P R (F )
F S S (M )
F S S (F )
IB H (M )
IB H (F )
0 1 2 3 4 6 8 1 0 1 4 3 0 6 00
1 0
2 0
3 0
T im e P o s t- In fe c tio n (D a y s )
IL-7
(p
g/m
l)
P R (M )
P R (F )
F S S (M )
F S S (F )
IB H (M )
IB H (F )
0 1 2 3 4 6 8 1 0 1 4 3 00
1 0 0 0
2 0 0 0
3 0 0 0
4 0 0 0
T im e P o s t- In fe c tio n (D a y s )
IL-8
(p
g/m
l)
P R (M )
P R (F )
F S S (M )
F S S (F )
IB H (M )
IB H (F )
0 1 2 3 4 6 8 1 0 1 4 3 0 6 00
1 0
2 0
3 0
T im e P o s t- In fe c tio n (D a y s )
IFN
- a (
pg
/ml)
P R (M )
P R (F )
F S S (M )
F S S (F )
IB H (M )
IB H (F )
0 1 2 3 4 6 8 1 0 1 4 3 0 6 00
2 0
4 0
6 0
8 0
T im e P o s t- In fe c tio n (D a y s )
IFN
- g (
pg
/ml)
P R (M )
P R (F )
F S S (M )
F S S (F )
IB H (M )
IB H (F )
0 1 2 3 4 6 8 1 0 1 4 3 0 6 00
1 0 0 0
2 0 0 0
3 0 0 0
4 0 0 0
T im e P o s t- In fe c tio n (D a y s )
MC
P-1
(p
g/m
l)
P R (M )
P R (F )
F S S (M )
F S S (F )
IB H (M )
IB H (F )
SouthernResearch
UBlityofCynomolgusMacaquesinZikaVaccineDevelopmentØ ImmunogenicityandEfficacy
Ø VaccineSafety
-Neurovirulence
-TissueTropism
Ø Dose-rangingandImmuneDuraBonstudy
Ø Cross-protecBonstudy
SouthernResearch
Summary
ü Cynomolgus macaques are susceptible to Zika infection ü PRVABC59 induced viremia detectable up to day 10, with peak
viral load at 2–3 days p.i. An intermittent recurrent viremia spike was observed on day 30 with titers reaching 2.5 x 103 genomes/mL
ü The Nab responses were found to be robust with the PRVABC59 strain which correlates with the observation made with serum viral loads
ü We found high levels as compared to the control of PDGF, IP-10, VEGF, RANTES, MIP1A, MIP1B, MCP-1, IL-1Ra, IFN-alpha with the higher dose of the PRVABC 59 strain
MCM could serve as an alternative model to study Zika infection and vaccine development
SouthernResearch
Acknowledgements
We would like to acknowledge the source of our ZIKV FSS13025 stock received from UTMB, Galveston National Laboratory, 301 University Boulevard, Galveston, TX 77550, USA; and ZIKV PRVABC59 from the CDC, 3156 Rampart Road, Fort Collins, CO 80526, USA.
FundingandContribuBngScienBsts
Southern Research Institute, Frederick, MD. (IR&D)
Scott Goebel Beth Snyder Kevin Walters Alison Gast Kimberly Hagelin Anju Singh Timothy J. Sellati Raj Kalkeri Jonathan Rayner
SouthernResearch