Trial Objectives immunogenicity safety
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1 Optimal priming of poxvirus vector (NYVAC)-based HIV vaccine regimens requires 3 DNA injections. Long term Results of the EV03/ANRS Vac20 Phase I/II Trial. Yves Levy, Kim Ellefsen, Wolfgang Stöehr, Pierre-Alexandre Bart, Jean-Daniel Lelièvre, Odile Launay, Hans Wolf, Jonathan Weber, Genevieve Chêne, Giuseppe Pantaleo
2 Trial Objectives A Phase I/II trial to compare the immunogenicity and safety of 3 DNA-C prime followed by 1 NYVAC-C boost to 2 DNA-C prime followed by 2 NYVAC-C boosts in healthy volunteers at low risk of HIV infection
3 Clinical Trial Design Weeks Group 1 (n=74) Group 2 (n=73) DNA-C (4 mg) priming at week 0, 4 and 8 for group 1 at week 0 and 4 for group 2 NYVAC-C ( PFUs) boosting at week 24 for group 1 and at week 20 and 24 for group 2 Randomized trial with a parallel group design No significant differences between groups in demographic characteristics at entry Open to the participants and investigators but blind to laboratory personnel Attendance to clinics at least 14 occasions over 72 weeks
4 Endpoints Immunogenicity The presence of CD8/CD4 T-cell responses to Env plus at least one of the Gag, Pol, Nef peptide pools at weeks 26 or 28 (primary end point) Long term immunogenicity at weeks 48 and 72 Safety Grade 3 or above Local & Systemic adverse event Grade 3 or above other clinical or laboratory adverse event confirmed at examination or on repeat testing respectively Any event attributable to vaccine leading to discontinuation of the immunisation regimen
5 Solicited and Non-Solicited Local and Systemic Adverse Events
6 Peptides Pools for IFN-γ ELISpot Peptide pools: Overlapping peptides (15-mers with 11 aa overlap, n=474) spanning the entire Gag/Pol/Nef polygene, and the Env clade C of HIV-1 97CN54. Grouped in 8 pools: Gag1/Gag2/Gag/Pol Pol1/Pol2 Nef Env1/Env2 Responders were defined as: > 4xfold the negative control and > 55 SFU/10 6 cells
7 Proportion of Responders at Primary Endpoints (Week 26/28) ITT Analysis 3 x DNA n = 70 2 x DNA n = 70 Total n = 140 Response 64 (91%) 56 (80%) 120 (86%) Chi 2 Test: p = 0.053; Risk difference: 11.4% (95% CI %) PP Analysis 3 x DNA n = 67 2 x DNA n = 68 Total n = 135 Response 63 (94%) 55 (81%) 118 (87%) Chi 2 Test: p = 0.021; Risk difference: 13.1% (95% CI %)
8 Proportion of Responders at Week 26/28 per Peptide Group ITT Analysis 3 x DNA n = 70 2 x DNA n = 70 Total n = 140 Env 63/70 (90%) 54/69 (78%) 117/139 (84%) Gag/Pol/Nef 27/70 (39%) 17/70 (24%) 44/140 (31%) PP Analysis 3 x DNA n = 67 2 x DNA n = 68 Total n = 135 Env 62/67 (93%) 53/67 (79%) 115/134 (86%) Gag/Pol/Nef 26/67 (39%) 17/68 (25%) 43/135 (32%)
9 Primary Immunogenicity Endpoints (Env at least one Gag, Pol or Nef pools) ITT Analysis 3 x DNA n = 70 2 x DNA n = 69 Total n = 139 Response 26 (37%) 15 (22%) 41 (30%) Chi 2 Test: p = 0.047; Risk difference: 15.4% (95% CI %) Risk ratio: 1.7 (95% CI ) PP Analysis 3 x DNA n = 67 2 x DNA n = 67 Total n = 134 Response 25 (37%) 15 (22%) 40 (30%) Chi 2 Test: p = 0.059; Risk difference: 14.9% (95% CI %) Risk ratio: 1.7 (95% CI )
10 Magnitude of IFN-γ ELISpot Responses at Week 26/28 Overall (SFUs/10 6 cells) Week 26 Week 28 ITT Analysis 3 x DNA n=58 2 x DNA n=50 3 x DNA n=61 2 x DNA n=54 Mean (SD) 774 (622) 398 (318) 597 (519) 357 (319) Median (IQR; range) ( ; ) ( ; ) ( ; ) ( ; ) Mann- Whitney test p<0.001 p<0.001 Note: Sum of SFU/Mio cells from all peptide pools with a positive response per participant
11 Proportion of Env Responders at W48 and W72 P = Percentage of responders P = W48 W72 W48 W72 3xDNA 1x NYVAC 2xDNA 2x NYVAC
12 Proportion of Gag/Pol/Nef Responders at W48 and W72 30 Percentage of responders W48 W72 W48 W72 3xDNA 1x NYVAC 2xDNA 2x NYVAC
13 Magnitude of Responses at W48 and W72 (mean of responding pools per subjects) SFU/10 6 Cells W48 W72 W48 W72 3xDNA 1x NYVAC 2xDNA 2x NYVAC
14 EV03/ANRS VAC20: Preliminary Flow Cytometry Analysis 27 participants analyzed 14 participants within Gr#1 (3xDNA1xNYVAC) 13 participants within Gr#2 (2xDNA2xNYVAC); only 11 participants were considered in the present analyses
15 Functional Profile of HIV-Specific T-Cell Responses (Env/Gag/pol/Nef pools) CD4 T-cell responses CD8 T-cell responses GR#1 GR#2 GR#1 GR#2 Frequency of CD4 T-cells IFNγ IL-2 TNFα Gr#1 (3xDNA) Gr#2 (2xDNA) Frequency of CD8 T-cells IFNγ IL-2 TNFα Gr#1 (3xDNA) Gr#2 (2xDNA)
16 Distribution of HIV Regions Targeted by CD4 T-Cell Responses P* = NS Gr#1 N=42 responses P* Fischer test Gr#2 N=22 responses GAG POL NEF ENV Number of responses ENV Gr#1 Gag, Pol or Nef Gr#2 ENV Gag, Pol or Nef
17 Distribution of HIV Regions Targeted by CD8 T-Cell Responses P* = 0.01 Gr#1 N=22 responses P* Fischer test Gr#2 N=8 responses GAG POL NEF ENV Number of responses ENV Gr#1 Gag, Pol or Nef Gr#2 ENV Gag, Pol or Nef
18 Breadth of CD4 and CD8 T-Cell Responses (ICS assay, Env/Gag/pol/Nef pools) CD4 T-cell responses CD8 T-cell responses P= Number of subjects Number of subjects Gr#1 (3xDNA) Gr#2 (2xDNA) Number of responses Number of responses Responders were defined as: > 0.03% CD4Cyt or CD8CytT cells and 2x fold background
19 Total Magnitude of T-Cell Responses (Sum of Responding Pools) 2.0 P=0.02 Percentage of T-cells Mean Gr1 (3xDNA N=14 Gr2 (2xDNA) N=11
20 Conclusions Both the 3xDNA1xNYVAC and 2xDNA 2xNYVAC vaccine regimens are safe and well tolerated The DNA/NYVAC prime/boost vaccine combination is highly immunogenic (94% responders in the PP analysis) Optimal priming of poxvirus-based vaccine regimens requires 3 DNA injections: superior for both proportion of polyepitopic responders and magnitude of the response elicits sustained responses Extends the breadth of CD8 T cell responses
21 Acknowledgements Vaccine Immunotherapy Centre Div. of Immunology and Allergy CHUV Lausanne, Switzerland Giuseppe Pantaleo Pierre-Alexandre Bart Erika Castro David Bonnet Kim Ellefsen-Lavoie Alexandre Harari St. Mary Hospital, Imperial College London, UK Jonathan Weber Rebecca Chandler Lucy Garvey Ken Legg Ngaire Latch University of Regensburg, Germany Bernd Salzberger Hans Wolf Birgit Fritsch Falitsa Mandraka Gabriele Birkenfeld Caspar Franzen Josef Köstler Ralf Wagner Cochin, Paris, France Odile Launay Pierre Loulergue Yvette Henin Henri Mondor, Paris, France Yves Lévy Jean-Daniel Lelièvre Christine Lacabaratz Tenon, France Gilles Pialoux Marseille, France Isabelle Poizot-Martin Catherine Farnarier Toulouse, France Lise Cuzin Florence Nicot INSERM CTU U897, France Genevieve Chene Philippe Reboud Inga Tschöpe Carine Grondin Valérie Boilet MRC CTU, London, UK Sheena McCormack Abdel Babiker Wolfgang Stöhr Liz Brodnicki Patrick Kelleher Mary Rauchenberger Shabana Khan ANRS Jean-Francois Delfrassy Yves Levy Anna-Laura Ross Véronique Rieux EuroVacc Foundation Song Ding Sanofi Pasteur, France Jim Tartaglia Claude Meric European Commission ANRS CAVD & All Study Volunteers Collaboration for AIDS Vaccine Discovery (CAVD) Nina Russell Jose Esparza
22 Participating Institutions Clinical Centers CHUV (Switzerland) Imperial College (UK) UREG (Germany) ANRS Centers (France) Mondor Cochin Tenon Toulouse Marseille Principal Investigators Giuseppe Pantaleo Yves Levy Sponsors EuroVacc Foundation ANRS Trial Coordination MRC CTU INSERM U593/CTU Immunology Laboratories CHUV (ELISpot assay) Equipe INSERM U955 (Flow Cytometry)
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