Memory HIV-specific T cell immune responses elicited by <i>A46R</i> deletion mutant in the spleen of BALB/c mice after prime/boost immunization.

<p>(A) Magnitude of the vaccine-specific CD4 or CD8 T cell responses. The HIV-specific CD4 or CD8 T cells were measured 53 days after the last immunization by ICS assay following stimulation of splenocytes derived from immunized animals (n=4) with the different HIV peptide pools. The total value in each group represents the sum of the percentages of CD4⁺ or CD8⁺ T cells secreting IFN-γ and/or IL-2 and/or TNF-α (CD4) or CD107a and/or IFN-γ and/or IL-2 and/or TNF-α (CD8) against all HIV peptide pools. All data are background-subtracted. *** <i>p</i><0.001. <i>p</i> value indicates significantly higher responses compared to parental group or between DNA-C/NYVAC-C and DNA-C/NYVAC-C-ΔA46R immunization groups. (B) Functional profile of the memory HIV-specific CD8 T cell response in the different immunization groups. The possible combinations of the responses are shown on the <i>x</i> axis, whereas the percentages of the functionally distinct cell populations within the total CD8 T cell population are shown on the <i>y</i> axis. Combinations that did not contribute significantly to the functional profile are not shown. Responses are grouped and colour-coded on the basis of the number of functions. *** <i>p</i><0.001. <i>p</i> values indicate significantly higher responses compared to DNA-C/NYVAC-C immunization group. (C) Phenotypic profile of memory HIV-specific CD8 T cells. Representative FACS plots showing the percentage of Env-specific CD8 T cells with central memory (TCM; CD127⁺CD62L⁺), effector memory (TEM; CD127⁺CD62L^-) or effector (TE; CD127^-CD62L^-) phenotype.</p

Deletion of <i>A46R</i> gene from NYVAC-C enhances innate immune responses.

<p>Human macrophages were mock-infected (0) or infected with NYVAC-WT, NYVAC-C or NYVAC-C-Δ46R (1 or 5 PFU/cell). 24 hours later, cell-free supernatants were collected to quantify the concentrations of TNF and IL-6 by bioassay and of IL-8 by ELISA. Data are means ± SD of duplicates and are representative of three independent experiments. * <i>p</i><0.05, ** p<0.005.</p

Adaptive HIV-specific T cell immune responses elicited by <i>A46R</i> deletion mutant in the spleen of BALB/c mice in heterologous prime/boost immunization protocol.

<p>(A) Magnitude of the vaccine-specific CD4 or CD8 T cell response. The HIV-specific CD4 or CD8 T cells were measured 10 days after the last immunization by ICS assay following stimulation of splenocytes derived from immunized animals (n=4) with the different HIV peptide pools. The total value in each group represents the sum of the percentages of CD4⁺ or CD8⁺ T cells secreting IFN-γ and/or IL-2 and/or TNF-α (CD4) or CD107a and/or IFN-γ and/or IL-2 and/or TNF-α (CD8) against all HIV peptide pools. All data are background-subtracted. *** <i>p</i><0.001. <i>p</i> value indicates significantly higher responses compared to parental group or between DNA-C/NYVAC-C-ΔA46R and DNA-C/NYVAC-C immunization groups. (B) Flow cytometry profiles of vaccine-induced CD4 or CD8 T cell responses against Env pool. (C) Functional profile of the adaptive HIV-specific CD4 or CD8 T cell response in the different immunization groups. The possible combinations of the responses are shown on the <i>x</i> axis, whereas the percentages of the functionally distinct cell populations within the total CD4 or CD8 T cell population are shown on the <i>y</i> axis. Combinations that did not contribute significantly to the functional profile are not shown. Responses are grouped and colour-coded on the basis of the number of functions. The non-specific responses obtained in the control group DNA-ϕ/NYVAC-WT were subtracted in all populations. ** p<0.005, *** <i>p</i><0.001. <i>p</i> values indicate significantly higher responses compared to DNA-C/NYVAC-C immunization group.</p