Predicted target genes for all the differentially expressed miRNAs.

<p>Graphs represent the percentage of predicted target genes grouped by pathways for the differential miRNA expression pattern in comparisons with resting CD8+ T cell samples (A) and with stimulated CD8+ T cell samples (B). <i>VP</i>, <i>viremic progressors; EC</i>, <i>elite controllers; ART</i>, <i>patients on antiretroviral therapy; HIV-</i>, <i>uninfected donors; VC</i>, <i>viremic controllers</i>.</p

Validation analysis of the miRNA expression profiles in CD8+ T-cells through individual RT-qPCR assays, normalized to endogenous control miRNAs.

<p>Graphs represent expression levels of: A) hsa-miR-149-3p; B) hsa-miR-4484; C) hsa-miR-4485; and D) hsa-miR-155-5p. One-way analysis of variance (ANOVA) tests were performed for global comparisons and Turkey post-hoc tests for pair comparisons. Bars represent standard error means (SEM); *, p<0.05. <i>VP</i>, <i>viremic progressors; EC</i>, <i>elite controllers; ART</i>, <i>patients on antiretroviral therapy; HIV-</i>, <i>uninfected donors; VC</i>, <i>viremic controllers</i>.</p

Differential miRNA profile between stimulated and resting CD8+ T-cells.

<p>Graphs represent mean fold change of the differential miRNA expression between stimulated and resting CD8+ T-cells from: A) VP; B) EC; C) ART; D) HIV-; and E) VC. A fold change value of >±1.5 was considered. <i>VP</i>, <i>viremic progressors; EC</i>, <i>elite controllers; ART</i>, <i>patients on antiretroviral therapy; HIV-</i>, <i>uninfected donors; VC</i>, <i>viremic controllers</i>.</p

Differential miRNA profile between stimulated CD8+ T-cells.

<p>Graphs represent mean fold change of the differential miRNA expression between stimulated CD8+ T-cells from: A) EC vs VP; B) ART vs VP; and C) HIV- vs VP. A fold change value of >±1.5 was considered. <i>VP</i>, <i>viremic progressors; EC</i>, <i>elite controllers; ART</i>, <i>patients on antiretroviral therapy; HIV-</i>, <i>uninfected donors; VC</i>, <i>viremic controllers</i>.</p

Baseline characteristics of the study participants.

<p>Baseline characteristics of the study participants.</p

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