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

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Noël TORDO CANINE ADENOVIRUS- BASED VACCINES AGAINST RABIES 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 UMR Virologie Marion SZELECHOWSKI Annie FOURNIER Bernard KLONJKOWSKI Marc ELOIT

previous papers…

G PV E1 0 E3 + E1 + left ITR right ITR E3 0 E1 0 E1 + E3 + GFP Replicative Cav2 Cav-G R + Non Replic. Cav2 Cav-G R 0 Non Replic. Human Ad5-G R 0 Replicative Cav2 Cav-GFP R + Wild-type Cav2 Recombinant canine adenovirus (Cav2) E3 + E1 + G PV E3 + G PV

Ad5-G R 0 Cav2-G R + Cav2-G R 0 Cav2 Ad5 G R 0 Cav2 G R + Cav2 G R 0 Cav2Mpurif.GNI % % % % Ad5-G R 0 Cav-G R + Cav-G R 0 Cav2 Expression of recombinant Gpv in CHO-CAR cells (24h; 10 TCID 50 ) Western blot Polyclonal G 4846 IF MAb G D % Flow cytometry MAb G D1

Total Infectious Units/ml Hours DK DK-E1 293 CRFK Multiplication of CaV2-derived viruses in various cell lignes (MOI = 1) DK Hours Total Infectious Units/ml Cav2-GFP R + Cav2-G R + Cav2 Hours DK-E1 Total Infectious Units/ml Cav2-GFP R + Cav2-G R + Cav2 dog cells other cells

l I.M. immunization (tibialis muscle) 10 7 TCID 50 Cav2 in 50 l / mouse l 27 day (VNAbs) l 48 days (VNAbs + T cell response) Immunogenicity of recombinant Cav2 in C57BL/6 mice by IM route ( 10 7 TCID 50 )

Ad5-G R 0 Virus Neutralizing Antibodies (VNA)Proliferative Index (G protein) Cav-G R 0 Cav-G R + Cav2Ad5-G R 0 Cav-G R 0 Cav-G R + Cav2 Immunogenicity of recombinant Cav2 by IM route ( 10 7 TCID 50 ) individual results

D27D48 VNA GMTRange VNA GMT Range Ad5-GR 0 316, , Cav-G R 0 58,2347,6-72,346,1823,1-72,4 Cav-G R + 56,3344,2-68,968, Cav200 D48 Proliferative index Range Ad5-G 0 2,231,5-3 Cav-G R 0 1,941,1-2,7 Cav-G R + 1,981,7-4,6 Cav21 Immunogenicity of recombinant Cav2 by IM route ( 10 7 TCID 50 ) Virus Neutralizing Antibodies (VNA)Proliferative Index (G protein)

l 21 day (VNAbs) l 25 day (challenge) Protection of recombinant Cav2 IM ( TCID 50 ) or oral ( TCID 50 ) routes TCID TCID 50

VaccinationRoute VNA GMT (IU) RangeSurvivors Cav-G R+IM45, /9 Cav-G R+oral1,640,38-7,57/9 Cav-GFP R+ IM00/5 Cav-GFP R+ oral00/5 Control00/3 Protection of recombinant Cav2 IM ( TCID 50 ) or oral ( TCID 50 ) 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)

Experiments in dogs in progress… Collaboration with the AFSSA Nancy Florence CLIQUET Jacques BARRAT

CONCLUSIONS. ã Cav2 grow well in canine cell lines and produce significant quantities of recombinant rabies G protein. ã Canine adenovirus (Cav2) -derived viruses (replicative and non replicative) expressing the rabies (PV strain) G protein have been produced. ã Cav2 are immunogenic (humoral and cellular responses) by both IM and oral routes. ã Dog experiments are in progress…. ã 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