Avian influenza heterologous prime-boost vaccination strategies. Michel BUBLOT Boehringer Ingelheim AH R&D (France)
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3 Avian influenza heterologous prime-boost vaccination strategies Michel BUBLOT Boehringer Ingelheim AH R&D (France)
4 Avian influenza vaccines - History 1970s Inactivated Fowlpox Rev. Gen. NDV HVT Baculovirus vaccine vector Inactivated vector vector (BEST) vaccine subunit H5, H7, H9 H5 H5 R&D R&D H5
5 Weaknesses of current AI vaccines Broadness of protection Vacc. 21DO EGYcls/09 EGYvar/08 H5N2 (A) 100% 40% H5N2 (B) 100% 60% Re5 H5N1 100% 100% - 0% 0% Grund et al. (2011) Vaccine 29, 5567 Maternally-derived antibody (MDA) interference Vacc. 3DO MDA HI titer EGYcls/09 H5N2 (A) % H5N2 (A) % - - 0% - + 0% Abdelwhab et al. (2011) Vet Microbiol 155, 13
6 How to improve protection with existing vaccines? Use two different vaccines in a heterologous prime-boost vaccination scheme
7 Heterologous Prime-Boost D0 1. Hatchery vaccination with Fowlpox-AI vaccine D Field vaccination with inactivated vaccine Why «heterologous»? 2 vaccine types 2 different HAs
8 Inactivated vaccine 3 important factors: Quality and Quantity of antigen Emulsion Vaccine strain All contribute to broadness of protection But: Better immunogenicity if admin. at 10 days of age MDA interference (higher if same vaccine used in breeders) Vaccine strain may sometimes need to be updated
9 Fowlpox-AI vector vaccine TROVAC -AIV H5 vaccine: vfp89: HA gene from A/turkey/Ireland/1378/1983 Licensed in 1998 in the U.S.A. > 9 B doses used Hatchery vaccination Broad protection : H5N2 Mexican LPAI & HPAI + old HP H5 HPAI H5N1 clades 0, 1, 2.2, 2.5 ( ) Immunogenic in ducks But: Poor protection against variant H5 Anti-AI MDA interference Ck/Hong Kong/156/97 * CK/Hong Kong/915/97 Hong Kong/483/97 CK/Hong Kong/220/97 CK/Hong Kong/728/97 CK/Italy/1485/97 Peking Duck/Singapore/645/97 TK/England/50-92/91 * DK/Hong Kong/342/78 DK/Ireland/113/83 TK/Ireland/1378/83 * CK/Scotland/59 * Tern/South Africa/61 * CK/Pennsylvania/1/83 * TK/WI/68 TK/Ontario/7732/66 * Emu/Tx/39442/93 * CK/Pennsylvania/13609/93 Ruddy Turnstone/DE/244/91 CK/Mexico/ /94 CK/Queretaro/ /95 * H5N1 Eurasian North American Swayne et al. (2000) Vaccine 18:1088
10 Prime-boost in Mexican field situation (LPAI H5N2) Seroconversión a IA* Farm 1 Farm 2 Título geométrico promedio sem 7 sem 6 sem 7 sem 6 sem 7 sem sem 7 sem sem 7 sem 6 sem 7 sem D0 + RecombD0 Recomb+Emulsión EmulsiónD15 D15 *Granja 1 vs granja 2; Recombinante 1día de edad 1/1-sc; Emulsión 15 días edad (Arriola et al., 1999) 0.5ml/sc ; título geométrico promedio HI; serología a 6 y 7 semanas de edad. Higher and more homogenous HI titers
11 g10) - H5N9 antigen Prime-Boost immunogenicity D28 2,1 1,8 1,5 1,2 0,9 0,6 0,3 0, HI t it ers ( log10 ) - H5N8 ant igen H5N8 antigen HI titers (log10) - H5N1 antigen H5N1 antigen 2,1 2,1 V1 1,8 1,8 V2 1,5 V1 1,5 V2 I n ac D0 +D2 1 Inac D0+D21 V1 1,2 1,2 vfp89+i nac D21 0,9 0,9 vfp8 9 +in ac D2V2 1 0,6 0,6 0,3 0,3 0,0 0,0 D0 D21 D28 D42 D0 D0 D21 D21 D28 D28 D42 D0 D42 D21 D28 D42 D42 Inac D0+D21 HI titers (log10) vfp89+inac - H5N9 antigen D21 vfp89+inac D21 EOPS H5N9 antigen Breeder D0 D21 FP/H5N9 H5N9 It H5N9 It FP/H5N9 vfp89 H5N9 It D0 D21 D28 D42 Prime-boost in birds with FP & AI MDA: Higher and broader HI titers than 2x inactivated Richard-Mazet et al. (2014) Vet. Res. 45:107 Inac D0+D21 vfp89+i nac D21
12 MDA interf. on inact. vaccine Breeder D0 D14 SPF - H5N9 It AI (3x H5N9) - H5N9 It Richard-Mazet et al. (2014) Vet. Res. 45:107 2,7 9 2,4 8 2,1 7 1,8 6 1,5 5 1,2 4 0,9 3 0,6 2 0, MDA interference on Inact. (H5N9) 2,7 2,4 2,1 V 1,8 1,5 1,2 0,9 0,6 0,3 0 D0 D14 D28 D42 MDA interference on Inact. (H5N8) MDA interference on Inact. (H5N1) H5N9 H5N8 H5N1 G0c-H5N9 V G1c-H5N9 2,7 2,4 2,1 1,8 1,5 1,2 0,9 0,6 0,3 0 D0 D14 D28 D42 G0c-H5N8 V G1c-H5N8 D0 D14 D28 D42 Clear interference of AI MDA on inact. at D14 G0c-H5N1 G1c-H5N1
13 ) Prime-Boost immunog. in MDA+ D42 vfp89 overcomes G4a-H5N1 interference MDA on interference inact. (H5N9) on prime-boost MDA interference (H5N8) on prime-boost (H5N1) 2,7 9 2,4 8 2,1 7 1,8 6 1,55 1,2 4 0,9 3 0,6 2 0,3 1 0,00 V1 V2 G0a-H5N1 G1a-H5N1 G3a-H5N1 H5N9 G5a-H5N1 2,7 2,4 2,1 1,8 1,5 1,2 0,9 0,6 0,3 0,0 D0 D14 D28 D42 Breeder D0 D14 SPF - H5N9 It AI 3x - H5N9 It AI 3x vfp89 H5N9 It H5N8 2,7 2,4 2,1 V1 1,8 V2 Inact (SPF) 1,5 1,2 Inact (M DA H5N9) 0,9 0,6 vfp89+inact (M DA H5N9) 0,3 0,0 D0 D14 D28 D42 H5N1 Inact (SPF) V2 Inact (MDA H5N9) vfp89+inact (MDA H5N9) D0 D14 D28 D42 TROVAC priming overcomes MDA interference on inactivated vaccine V1 Richard-Mazet et al. (2014) Vet. Res. 45:107 Inact Inact H5N9) vfp89 (MDA
14 Prime-Boost (It-H5N9) protection D0 / D14: Vaccination of SPF & FP MDA+ chicks D28: chall. with A/ck/West Java (Spg)/29/2007 H5N1 Richard-Mazet et al. (2014) Vet. Res. 45:107 MDA V1 - D0 Vaccination V2 - D14 H5N8 HI titers D28 Protection (chall. at D28) D. Swayne /10 (0%) - vfp (100%) 1/10 (10%) FP - H5N9 inact. 5.1 (100%) 5/9 (56%) FP vfp89 H5N9 inact. 8.1 (100%) 9/9 (100%) Synergistic effect of prime-boost on HI titers and protection in birds with anti-vector (FP) MDAs
15 Prime-Boost (It-H5N9) protection D0 / D14: Vaccination of AI MDA+ chicks D28: chall. with A/ck/West Java (Spg)/29/2007 H5N1 Richard-Mazet et al. (2014) Vet. Res. 45:107 MDA V1 D0 V2 D14 H5N8 HI titer D28 Protection (chall. at D28) D. Swayne H5N (0%) 0/9 (0%) H5N9 vfp (56%) 0/9 (0%) H5N9 - H5N9 inact. 3.2 (44%) 4/9 (44%) H5N9 vfp89 H5N9 inact. 5.8 (100%) 7/9 (78%) Synergistic effect of prime-boost on HI titers and protection in birds with anti-ai MDAs
16 Prime-Boost (Re5) protection D0 / D21: vaccination of SPF chicks D42: Challenge with A/chicken/Egypt/1063/2010 (EGYvar) 6 log10 IN Vaccine Vaccine Clinical protection Shedding D0 D21 D42 2 dpch 4 dpch vfp89 Re5 25/25 (100%) 11/25 (44%) 12/25 (48%) - Re5 14/15 (93%) 14/15 (93%) 15/15 (100%) - - 0/25 (0%) 25/25 (100%) - Prime-Boost : full clinical protection & better reduction of shedding
17 Prime-Boost (Re5) serology D0 / D21: vaccination of SPF chicks D42: Challenge with A/chicken/Egypt/1063/2010 (EGYvar) 6 log10 IN Vaccine D0 Vaccine D21 Re5 Ag HI titer (log2) - D42 EGYvar Ag vfp89 Re (25/25) 3.7 (16/25, 64%) - Re5 9.3 (15/15) 2.2 (3/15, 20%) (0/25) - Prime-Boost : Broader HI titers
18 Prime-Boost (Re5) protection D0 / D10: vaccination of SPF chicks D24: Chall. with A/chicken/Egypt/1063/2010 (EGYvar) 6 log10 IN/oral Vaccine Vaccine Clinical protection Shedding D0 D10 D24 Ct<35 Ct<40 vfp89 Re5 15/15 (100%) 0/15 (0%) 1/15 (7%) - Re5 15/15 (100%) 2/15 (13%) 6/15 (40%) *Number of live birds/total having at least one positive swabs at 3, 5, 7 or 10 days Prime-Boost : Better reduction of shedding
19 Prime-Boost (Re5) serology D0 / D10: vaccination of SPF chicks D24: Chall. with A/chicken/Egypt/1063/2010 (EGYvar) 6 log10 IN/oral Vaccine D0 Vaccine D10 Re5 Ag HI titer (log2) D24 EGYvar Ag vfp89 Re5 5.4 (15/15) 0.7 (8/15, 53%) - Re5 5.5 (15/15) 0.1 (1/15, 7%) Prime-Boost : Broader HI titers
20 Prime-Boost (It-H5N9) in ducklings 17-day-old Pekin ducks D0 & D24: priming with It-H5N9 or vfp-h5 (H5N1 HA) D24: boost with Italy- H5N9 inact vaccine D0, D13, D24, HI titers D37: (log2) - prechall. HI titers - H5N9 (log2) antigen pre- & post-vaccination HI titers (log2) - prechall. - H5N1 antigen Inact 2x FP-Inact V1 H5N9 antigen V2 Pre-V 13d pv1 24d pv1 13d pv Inact 2x FP-Inact V1 H5N1 cl. 1 antigen V2 Pre-V 13d pv1 24d pv1 13d pv2 Steensels et al. (2009) Vaccine 27, Prime Boost Broader antibody response
21 Heterologous prime-boost principles 1. Different antigen presentation Fowlpox: Cell-Mediated Immunity (Hghihghi et al. (2010) Clin Vacc Immunol 17, ) Inactivated: Humoral Immunity 2. Boost directed to protective antigen (HA) HA NP M1, M2 HA NA 3. Boost with a different HA to increase response against conserved epitopes (broader response) A AB B AC BC ABC B + B B mainly A + B AB + ABC (2 shots of inact.) (prime-boost) C Bublot et al. (2008) Res. Sign Post 37/661(2),
22 Feb 26, 2010 April 2014
23 Conclusion on prime - boost Optimal immune response protection + shedding Broader antibody response against various Ags Overcomes AI MDA interference on inactivated vaccine Multi-species Compatible with HVT vector vaccines Perspectives: FP + BEST Prime-Boost DIVA (NP ELISA)
24 Acknowledgements SEPRL: J. Beck, D. Suarez, D. Swayne IZSVe: M.S. Beato, C. Terregino, I. Capua VAR: M. Steensels, S. Van Borm, B. Lambrecht, Th. van den Berg B.I. R&D: A. Richard-Mazet, S. Chanavat-Bizzini, F.-X. le Gros, S. Goutebroze, M. Duboeuf, S. Brunet, A. Dancer, V. Cozette, N. Pritchard
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