Adverse events related to vaccination description and proportion of volunteers suffering any side effects.

<p>Adverse events related to vaccination description and proportion of volunteers suffering any side effects.</p

Humoral responses.

<p>A: Total IgG binding antibody titers against HIV-1 gp120 (BX08). B: BX08 neutralization ID50 titers. C: Correlation between BX08 binding IgG and neutralizing ID50 titers. D: Total IgG binding titers against VACV proteins. E: VACV neutralization ID50 titers. The frequency of responders and the mean titers at the different time points are shown in each graph. Dashed line represents the threshold considered as positive response. Statistical differences were evaluated by one way ANOVA test (using the linear model log10(y) ~ x_patient + x_week + epsilon) followed by Tukey's honest significant difference criterion. *p<0.05, **p<0.01, *** p<0.005 (***4). Pearson’s correlation coefficient (r value) was calculated between BX08 binding IgG and neutralizing ID50 titers.</p

Serum neutralizing activity against HIV-1 BX08 virus at 2, 4 and 12 weeks after receiving the boost.

<p>Data are shown as the reciprocal dilution giving 50% and 80% neutralization (ID50 and ID80 titers). Reciprocal serum ID50 and ID80 values ≥1000 are highlighted in red, ≥400 and <1000 in orange, ≥200 and <400 in dark yellow, ≥90 and <200 in light yellow, and <90 in white. The given reciprocal titers correspond to 1/dilution of serum.</p

Frequency, function and phenotype of HIV-specific T cell responses.

<p>A: Percentage of responders with positive ICS responses against Env+Gag+GPN at the different time points (left panel) and distribution of the CD4+ and CD8+ T cells by antigen (right panel). B: Functional profile of vaccine-induced T cells. The quality of the HIV-specific CD4 or CD8 T cell response is characterized by the proportion of cells making every possible combination of the measured cytokines: IFN-γ (I); IL-2 (2); TNF-α (T) and CD107a (C). Responses are grouped and colour coded on the basis of the number of functions. The bar charts show the mean values and interquartile ranges (IQR) and the pie charts show the average proportion of the HIV-specific CD4 or CD8 T cell responses according to the functions at weeks 0, 2, 4 and 12. “+”distributions that are different from the earliest time point (W0) within each category at p<0.05 (Student's T test). C: Phenotype of vaccine-induced T cells. The graphic represents the distribution of the responding HIV-specific CD4 and CD8 T cells at any time point based on CCR7 expression in combination with CD45RA within the Naïve (CD45RA+ CCR7+), T central memory (TCM: CD45RA- CCR7+), T effector memory (TEM: CD45RA- CCR7-) or terminally differentiated T effector memory (TEMRA: CD45RA+ CCR7-) phenotypes. Statistical differences were determined using ANOVA test (using the linear model y ~ x_patient + x_cellType + epsilon) followed by Tukey's honest significant difference criterion. *p<0.05, ***p<0.005.</p

ELISPOT results.

<p>Magnitude of HIV-1-specific T cell responses measured by IFN-γ-based ELISPOT is shown. A) Total responses, represented as the sum of positive responses to Gag, GPN and Env peptide pools; B) T-cell responses to Gag peptide pools; C) Positive responses to GPN peptide pools, and D) T-cell responses to Env peptide pools. The graphs show the frequency of HIV-1-specific T cell responses by SFC/106 PBMC at different time-points (-3, w0, w2, w4 and w12). Week -3 corresponds to w48 of follow-up of RISVAC02 clinical trial. Median and IQR are represented in all the graphs for the different time-points evaluated.</p

Frequency, function and phenotype of VACV-specific CD8 T cell responses.

<p>A: Percentage of responders with positive ICS responses against MVA infected cells at the different time points. The equality of proportion between groups was determined using the function prop.test. *p = 0.017. B: Functional profile of VACV-specific CD8 T cells. The quality of the VACV-specific CD8 T cell response is characterized by the proportion of cells making every possible combination of the measured cytokines: IFN-γ (I); IL-2 (2); TNF-α (T) and CD107a (C). Responses are grouped and colour coded on the basis of the number of functions. The bar charts show the mean values and interquartile ranges (IQR) and the pie charts show the average proportion of the VACV-specific CD8 T cell responses according to the functions at weeks 0, 2, 4 and 12. “+”distributions that are different from the earliest time point (W0) within each category at p<0.05 using Student's T test or “#” Wilcoxon signed rank test. C: Phenotype of VACV-specific CD8 T cells. The graphic represents the distribution of the VACV-specific CD8 T cells at any time point based on CCR7 expression in combination with CD45RA within the Naïve (CD45RA+ CCR7+), T central memory (TCM: CD45RA- CCR7+), T effector memory (TEM: CD45RA- CCR7-) or terminally differentiated T effector memory (TEMRA: CD45RA+ CCR7-) phenotypes. Statistical differences were determined using ANOVA test (using the linear model y ~ x_patient + x_cellType + epsilon) followed by Tukey's honest significant difference criterion. *p<0.05.</p

Disposition of participants and study flow chart.

<p>A total of 24 HIV-negative volunteers were vaccinated with MVA-B in the RISVAC02 study. Four years later, 13 of these volunteers were recruited to receive a single intramuscular boost of MVA-B vaccination in the RISVAC02boost clinical trial. Adverse Events (AEs), laboratory analyses and immunogenicity endpoints were tested as scheduled in the flow chart.</p

Volunteer characteristics and adverse events reported per volunteers according to relationship between vaccination, distribution and gradation.

<p>Volunteer characteristics and adverse events reported per volunteers according to relationship between vaccination, distribution and gradation.</p