H. Kibuuka, N. M. Berkowitz, M. Millard, M. E. Enama, A. Tindikahwa et al., Safety and immunogenicity of Ebola virus and Marburg virus glycoprotein DNA vaccines assessed separately and concomitantly in healthy Ugandan adults: a phase 1b, randomised, double-blind, placebo-controlled clinical trial, The Lancet, vol.385, issue.9977, pp.1545-54, 2015.

A. Guardo, C. E. Gómez, V. Díaz-brito, J. Pich, J. A. Arnaiz et al., Safety and vaccine-induced HIV-1 immune responses in healthy volunteers following a late MVA-B boost 4 years after the last immunization, PLoS One, vol.12, issue.10, 2017.

Y. Lévy, R. Thiébaut, M. Montes, C. Lacabaratz, L. Sloan et al., Dendritic cellbased therapeutic vaccine elicits polyfunctional HIV-specific T-cell immunity associated with control of viral load, Eur J Immunol, vol.44, issue.9, pp.2802-2812, 2014.

A. Luabeya, B. Kagina, M. D. Tameris, H. Geldenhuys, S. T. Hoff et al., Firstin-human trial of the post-exposure tuberculosis vaccine H56:IC31 in Mycobacterium tuberculosis infected and non-infected healthy adults, Vaccine, vol.33, issue.33, pp.4130-4170, 2015.

H. Horton, E. Thomas, J. Stucky, I. Frank, Z. Moodie et al., Optimization and Validation of an 8-Color Intracellular Cytokine Staining (ICS) Assay to Quantify Antigen-Specific T Cells Induced by Vaccination, J Immunol Methods, vol.323, issue.1, pp.39-54, 2007.

D. Rosa and S. C. , Vaccine applications of flow cytometry, Methods, vol.57, issue.3, pp.383-91, 2012.

L. Lin, G. Finak, K. Ushey, C. Seshadri, T. R. Hawn et al., COMPASS identifies T-cell subsets correlated with clinical outcomes, Nat Biotechnol, vol.33, issue.6, pp.610-616, 2015.

G. J. Churchyard, C. Morgan, A. E. Hural, J. Graham, B. S. Moodie et al., A phase IIA randomized clinical trial of a multiclade HIV-1 DNA prime followed by a multiclade rAd5 HIV-1 vaccine boost in healthy adults (HVTN204), PLoS ONE, vol.6, issue.8, p.21225, 2011.

F. Y. Tung, J. K. Tung, S. Pallikkuth, S. Pahwa, and M. A. Fischl, A therapeutic HIV-1 vaccine enhances anti-HIV-1 immune responses in patients under highly active antiretroviral therapy, Vaccine, vol.34, pp.2225-2257, 2016.

H. E. Janes, K. W. Cohen, N. Frahm, D. Rosa, S. C. Sanchez et al., Higher T-Cell Responses Induced by DNA/rAd5 HIV-1 Preventive Vaccine Are Associated With Lower HIV-1 Infection Risk in an Efficacy Trial, J Infect Dis, vol.215, issue.9, pp.1376-85, 2017.

S. A. Kalams, S. D. Parker, M. Elizaga, B. Metch, S. Edupuganti et al., Safety and Comparative Immunogenicity of an HIV-1 DNA Vaccine in Combination with Plasmid Interleukin 12 and Impact of Intramuscular Electroporation for Delivery, J Infect Dis, vol.208, issue.5, pp.818-847, 2013.

L. Richert and R. Thiebaut, BiostatisticsBioinformatics Division of the ANRS Vaccine Research Institute. Is the current use of « positivity » thresholds meaningful for evaluating HIV-vaccine immunogenicity endpoints?, AIDS, vol.27, issue.8, pp.1362-1367, 2013.

A. Y. Liu, D. Rosa, S. C. Guthrie, B. L. Choi, R. Y. Kerubo-bosire et al., Lohman-Payne B High background in ELISpot assays is associated with elevated levels of immune activation in HIV-1-seronegative individuals in Nairobi, Immun Inflamm Dis, vol.6, issue.3, pp.392-401, 2018.

L. F. Su, B. A. Kidd, A. Han, J. J. Kotzin, and M. M. Davis, Virus-specific CD4+ memory phenotype T cells are abundant in unexposed adults, Immunity, vol.38, issue.2, pp.373-83, 2013.

P. B. Gilbert, A. Sato, X. Sun, and D. V. Mehrotra, Efficient and robust method for comparing the immunogenicity of candidate vaccines in randomized clinical trials, Vaccine, vol.27, issue.3, pp.396-401, 2009.

R. Thiébaut, H. Jacqmin-gadda, G. Chêne, C. Leport, and D. Commenges, Bivariate linear mixed models using SAS proc MIXED, Computer Methods and Programs in Biomedicine, vol.69, issue.3, pp.249-56, 2002.

Z. Moodie, A. J. Rossini, M. G. Hudgens, P. B. Gilbert, S. G. Self et al., Statistical evaluation of HIV vaccines in early clinical trials, Contemporary Clinical Trials, vol.27, issue.2, pp.147-60, 2006.

L. Richert, E. Lhomme, C. Fagard, Y. Lévy, G. Chêne et al., Recent developments in clinical trial designs for HIV vaccine research, Hum Vaccin Immunother. 9 mars, vol.11, issue.4, pp.1022-1031, 2015.
URL : https://hal.archives-ouvertes.fr/hal-01164644

, VICI: acurate estimation of Vaccine Induced Cellular Immunogenicity by bivariate modelling

J. P. Ioannidis, Why most discovered true associations are inflated, Epidemiology, vol.19, issue.5, pp.640-648, 2008.

H. Jacqmin-gadda, S. Sibillot, C. Proust, J. Molina, and R. Thiébaut, Robustness of the linear mixed model to misspecified error distribution, Computational Statistics & Data Analysis, vol.51, issue.10, pp.5142-54, 2007.

C. Appendix and . Analyzes,

, Creteil, France) on cryopreserved PBMC. PBMC were rested overnight and then stimulated (6h, 37°C, 5% CO2) with HIV peptide pools (1µg/ml) in the presence of co-stimulatoy molecules (anti-CD28 and anti-CD49d antibodies (1µg/ml)) and a protein transport inhibitor (Golgi Plug (1 µl/ml)) (BD Biosciences, To assess antigen-specific T-cell responses, ICS assay was performed in a centralized laboratory

, Saint Aubin, France) and with fluorochrome-conjugated monoclonal antibodies (anti-CD3 Alexa700, anti-CD4 PE, and anti-CD8 efluor 780; all from BD Biosciences) for 15 min at room temperature. After fixation and permeabilization using Cytofix/Cytoperm kit (BD Biosciences) for 20 min and staining with anti-IFN-g PerCpCy5.5, -TNF-? PE-Cy7 and -IL-2 APC (all BD Biosciences) for 20 minutes at room temperature, PBMCs were re-suspended in Paraformaldéhyde 1% (BD Biosciences) and stored at 4°C until analysis, ) served as positive control. After stimulation, cells were stained for dead cells with an amine-reactive dye (LIVE/DEAD Aqua, Invitrogen, Life Technologies, vol.640, p.561

, At least 250,000 events gated on CD3 + were collected and analyzed using Boolean gating, and 405 nm) cytometer (BD Biosciences), analyzed using FlowJo software version 9, vol.9