Replicating measles-shiv vaccine induces long term preservation of central memory CD4 cells in the gut of macaques challenged with SHIV89.
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1 Replicating measles-shiv vaccine induces long term preservation of central memory CD4 cells in the gut of macaques challenged with SHIV89.6P Frédéric Tangy Viral Genomics and Vaccination Laboratory
2 Measles virus (Morbillivirus, Paramyxoviridae) nm N P C V M F H non-segmented RNA, negative polarity (16 kb) L
3 Measles live attenuated vector Edmonston HK-24 HA-28 Edmonston-Enders CE-6 CEF-13 Edmonston A CEF/85 32 C Schwarz One injection : TCID 5 (9-16 months) Long-lived protective immunity (NAb + CD8 + CD4) High genetic stability Safety and efficacy track record in billions of children Established logistics for production and distribution of MV vaccine worldwide, low cost of production Mass vaccination needed for foreseeable future N P M F H L T7 hh C V (cloned unpassaged Schwarz strain) T7t - Tangy et al. EP , filed May 22 - Tangy et al. EP , filed June 23 - Tangy et al. EP , filed December 26 - Combredet et al. 23, JVI, 77,
4 Recombinant MV vaccine targets To immunize children simultaneously against measles and other diseases HIV Arboviruses (DV, WN, YF, CHIKV ) Acute respiratory diseases (RSV, PIV-3, SARS, H5N1 ) Malaria MV-Schwarz MV-CS-falciparum MV-DV-EDIII em MV-DV-EDIII em MV-S MV-Ssol MV-prME-YFV Permeabilized MV-N1 MV-S-SARS sol MV-sE-WNV Not Permeabilized MV-H5 MV-S-SARS
5 Measles vector expresses HIV proteins and is immunogenic MVSchw-Gag HIV MVSchw-Gag (P1) MVSchw-Gag (P3) MVSchw Vero cells Gag p42 MVSchw-Env HIV HIV Mv2-gp16 V3 MV2-gp14 MV2-gp16 MV3-gp14 V3 MV2-gp14 MV2-gp16 MV3-gp14 V3 Mv2-gp16 V MV 6 P2 P5 MV-HIV vectors induce HIVspecific CD4/CD8 T cells and antibodies in mice and macaques, even in presence of preimmunity to MV Lorin et al. 24, JVI, 78, Lorin et al. 25, Vaccine, 23: e5 1e4 1e3 1e2 1e Months MV MV Ab Mice HIV Ab MV-HIV x2 1e4 1e3 1e2 1e5 Macaques 1e4 1e4 1e3 1e3 1e2 1e2 MV Ab 1 1 HIV Ab Months MV MV-HIV x 2
6 Macaques immunization and SHIV89.6P challenge - 12 macaques cynomolgus vaccinated with a mixture of recombinant MVSchw : MV-(Gag SIV -gp14 HIV ) + MV-( Tat HIV ) + MV-(Nef SIV ) (1 6 TCID5 each) - Intra-rectal challenge with 3 AID 5 SHIV 89.6p MV-SHIV MV-SHIV SHIV A MV MV SHIV B Months HIV γ-ifn Elispots/1 6 cells #5797 Env Env Gag Gag Env Tat d d15 M1 M #5986 Gag Gag Tat Gag Env d d15 M1 M #6165 Gag Env Tat d d15 M1 M2
7 Immunity is induced for long years MV 2 x MV-SHIV Sacrifice C Months 5.5 years SHIV response PBMC Spleen MV response Overnight stimulation with specific peptides γ-ifn Elispots > 2 x background ICS and FACS analysis confirmation
8 2-4 logs reduction of SHIV acute viral load 1,E+9 Viral load at challenge 12 Ab resp at challenge SHIV viral load 1,E+8 1,E+7 1,E+6 1,E+5 1,E+4 1,E+3 1,E+2 1,E+1 Vaccinated Anti-HIV gp12 Ab titer (%) Vaccinated 1,E Days post challenge Months post challenge 45 Cellular resp at challenge 1 Neut Ab resp at challenge Gag SFC/million cells Vaccinated Days post challenge 5% neutralization titer Months post vaccination Vaccinated Days post challenge
9 Reactivation of SHIV replication through BCG inoculation Lymphocytes counts / ml after challenge Vaccinees Days Months Years SHIV challenge BCG (1 7 pfu iv) Sacrifice
10 Vaccinated animals control SHIV viral load 1e8 1e7 SHIV viral load copies/ml 1e7 1e6 1e5 1e4 1e3 1e2 Vaccinated SHIV viral load copies/ml 1e6 1e5 1e4 1e3 1e Days post BCG inoculation Days post BCG inoculation
11 Reactivation of SHIV-specific T cells after BCG inoculation During BCG activation, a higher level of SHIV-specific T cells were raised in vaccinated animals as compared to controls HIV specific γ-ifn Elispots/ million cells Days post BCG inoculation Days post BCG inoculation Vaccinees BCG (PPD)-specific γ-ifn Elispots/million cells Days post BCG inoculation Days post BCG inoculation Vaccinees
12 Preservation of CD4 memory cells in the gut of vaccinees MV-SHIV SHIV BCG Sacrifice Years Five years after challenge, macaques were sacrificed and T lymphocytes populations were analyzed in blood, lymphoid organs and in the gut. Although no difference was observed in the level of circulating CD4 cells between groups, the CD4 central memory lymphocytes were clearly preserved in the gut of vaccinees. CD4 FL4-H: CD4 APC CD4 CD FL3-H: CD3 PerCP-Cy CD3 CD28 FL1-H: CD28 FITC Naïve Central memory Effector memory FL2-H: CD95 PE 1 4 CD95 Vaccinated Unchallenged % CD4 CM p =,43 IEL Duodenum p =.48 LPL Duodenum Same observation in intra-epithelial lymphocytes (IEL) from jejunum, ileon and colon
13 MV recombinant vector expressing HIV VLPs T7 hh l N C P V HIV-p55 Gag M F H HIV-gp16 V12 L t T7t A 2 µm B 2 nm C 1 µm MV-p55Gag MV-p55Gag MV-p55Gag D E 5 nm F 2 nm MV-p55Gag MV-p55Gag/Env MV-p55Gag/Env Guerbois et al. 29, Virology, 388,191 23
14 RMVHI Demonstrating safety and immunogenicity of recombinant MV-HIV vector in adult HIV-uninfected volunteers V Generation of a GMP lot of rmv-hiv to elicit CMI Identification of a suitable dose of recombinant vaccine Demonstration of an acceptable reactogenicity profile Characterization of potential virus shedding Evaluation of HIV-specific CD4/CD8/Ab responses raised in volunteers with MV preimmunity GSK-Bio (Rixensart), Institut Pasteur (Paris), St George s Hospital Medical School (London), Ghent University, CIC Cochin Pasteur (Paris), NIBSC (South Mims) Program started in 25
15 Acknowledgements Michèle Février Chantal Combredet Valérie Labrousse Mathilde Guerbois Mariana Mesel Claude Ruffié CPR, Bruno Hurtrel CEA, Roger Legrand Viral Genomics and Vaccination laboratory
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