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Noël TORDO CANINE ADENOVIRUS- BASED VACCINES AGAINST RABIES working together to stop the ongoing tragedy of rabies!

Noël TORDO

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CANINE ADENOVIRUS-BASED VACCINES AGAINST RABIES. Noël TORDO. working together to stop the ongoing tragedy of rabies!. Institut Pasteur, Paris, France Unit Antiviral Strategies Corinne JALLET Noël TORDO INRA-AFSSA-ENVA Maisons-Alfort, France UMR1161 - Virologie Marion SZELECHOWSKI - PowerPoint PPT Presentation

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Page 1: Noël TORDO

Noël TORDO

CANINE ADENOVIRUS-BASED VACCINES AGAINST RABIES

working together to stop the ongoing tragedy of rabies!

Page 2: Noël TORDO

Institut Pasteur, Paris, FranceUnit Antiviral Strategies

Corinne JALLETNoël TORDO

INRA-AFSSA-ENVA Maisons-Alfort, France UMR1161 - Virologie

Marion SZELECHOWSKIAnnie FOURNIER Bernard KLONJKOWSKIMarc ELOIT

Page 3: Noël TORDO

previous papers…

Page 4: Noël TORDO

G PV

E10

E3+E1+

left ITR right ITR

E30E10

E1+ E3+

GFP

Replicative Cav2Cav-G R+

Non Replic. Cav2Cav-G R0

Non Replic. HumanAd5-G R0

Replicative Cav2Cav-GFP R+

Wild-type Cav2

Recombinant canine adenovirus (Cav2)

E3+E1+

G PV

E3+

G PV

Page 5: Noël TORDO

Ad5-G R0Cav2-G R+Cav2-G R0 Cav2

Ad5G R0

Cav2G R+

Cav2G R0

Cav2Mpurif.G NI

Data.004

100 101 102 103 104

FL1-Height

M1M2

Data.002

100 101 102 103 104

FL1-Height

M1M2

Data.006

100 101 102 103 104

FL1-Height

M1

M2

Data.008

100 101 102 103 104

FL1-Height

M1

M295.68 %531.80

92.44 %396

2.08 %21.26

99.51 %1162.35

Ad5-G R0Cav-G R+Cav-G R0 Cav2

Expression of recombinant Gpv in CHO-CAR cells (24h; 10 TCID50)

Western blotPolyclonal G4846

IF MAb G D1

99.51 %1162.35Flow

cytometry MAb G D1

Page 6: Noël TORDO

0 24 48 72 9610 0

10 1

10 2

10 3

10 4

10 5

10 6

10 7

10 8

10 9

10 10

To

tal I

nfe

ctio

us

Un

its/

ml

Hours

DK

DK-E1

293

CRFK

Multiplication of CaV2-derived viruses in various cell lignes (MOI = 1)

DK

0 24 48 72 9610 0

10 1

10 2

10 3

10 4

10 5

10 6

10 7

10 8

10 9

10 10

Hours

To

tal I

nfe

ctio

us

Un

its/

ml

Cav2-GFP R+

Cav2-G R+

Cav2

Hours

DK-E1

To

tal I

nfe

ctio

us

Un

its/

ml

0 24 48 72 9610 0

10 1

10 2

10 3

10 4

10 5

10 6

10 7

10 8

10 9

10 10

Cav2-GFP R+

Cav2-G R+

Cav2

Cav2

dog cells

other cells

Page 7: Noël TORDO

I.M. immunization (tibialis muscle) 107 TCID50 Cav2 in 50 l / mouse 27 day (VNAbs) 48 days (VNAbs + T cell response)

Immunogenicity of recombinant Cav2 in C57BL/6 mice by IM route (107 TCID50 )

Page 8: Noël TORDO

0

1

2

3

4

5

Pro

lifer

ativ

e In

dex

1

10

100

1000

UI/m

l

Ad5-G R0

Virus Neutralizing Antibodies (VNA) Proliferative Index (G protein)

Cav-G R0 Cav-G R+ Cav2 Ad5-G R0 Cav-G R0 Cav-G R+ Cav2

Immunogenicity of recombinant Cav2 by IM route (107 TCID50 )

individual results

Page 9: Noël TORDO

D27 D48

VNA GMT RangeVNA GMT

Range

Ad5-GR0 316,77 101-546 194,35 76-504

Cav-G R0 58,23 47,6-72,3 46,18 23,1-72,4

Cav-G R+ 56,33 44,2-68,9 68,27 52-78

Cav2 0 0

D48

Proliferative index

Range

Ad5-G0 2,23 1,5-3

Cav-G R0 1,94 1,1-2,7

Cav-G R+ 1,98 1,7-4,6

Cav2 1

Immunogenicity of recombinant Cav2 by IM route (107 TCID50 )

Virus Neutralizing Antibodies (VNA) Proliferative Index (G protein)

Page 10: Noël TORDO

21 day (VNAbs) 25 day (challenge)

Protection of recombinant Cav2IM (7.106 TCID50 ) or oral (7.107 TCID50 ) routes

7.106 TCID50

7.107 TCID50

Page 11: Noël TORDO

Vaccination RouteVNA GMT

(IU)Range Survivors

Cav-G R+ IM 45,28 25-83 9/9

Cav-G R+ oral 1,64 0,38-7,5 7/9

Cav-GFP R+

IM 0 0/5

Cav-GFP R+

oral 0 0/5

Control 0 0/3

0

20

40

60

80

100

3 4 5 6 7 8 9 10 11 12 13 14 15 16 17

days post challenge

% s

urv

ival

Cav-G R+ (I.M)

Cav-G R+ (oral)

Cav-GFP R+ (I.M)

Cav-GFP R+ (oral)

Control

Protection of recombinant Cav2IM (7.106 TCID50 ) or oral (7.107 TCID50 ) routes

IM Cav-G R+

OR Cav-G R+

IM Cav-GFP R+

OR Cav-GFP R+

Control

Virus Neutralizing Antibodies (day 21) Challenge (day 25)

Page 12: Noël TORDO

Experiments in dogs in progress…

Collaboration with the AFSSA NancyFlorence CLIQUETJacques BARRAT

Page 13: Noël TORDO

CONCLUSIONS

•.

Cav2 grow well in canine cell lines and produce significant quantities of recombinant rabies G protein

•.

Canine adenovirus (Cav2) -derived viruses (replicative and non replicative) expressing the rabies (PV strain) G protein have been produced

•.

Cav2 are immunogenic (humoral and cellular responses) by both IM and oral routes

•. Dog experiments are in progress…

•.

IM and oral vaccination of mice with replicative Cav2 G viruses protect them against a peripheral challenge

Cav2 G viruses are promising tools for oral vaccination of dogs

Page 14: Noël TORDO