Fig. 4: Analysis of specific CD4⁺ and CD8⁺ T cell responses against S^WT, S^D, S^O and S^B in V_NLRs, V_NHRs and V_R subjects.

a, Representative flow cytometry dot plots of CD25 versus (vs) CD69 cells on gated CD4⁺ (left) and CD8⁺ (right) T cells from V_NLRs, V_NHRs and V_R subjects at month 6, after 24h incubation with medium (Unstimulated) or S^WT peptide megapools. b,c, Flow cytometry analysis showing the percentage of CD25⁺CD69⁺ cells in CD4⁺ T cells (b) and CD8⁺ T cells (c) from V_NLRs, V_NHRs and V_R subjects at T0 and 6M, 24h after incubation with S^WT, S^D, S^O or S^B. Below: percentages of subjects with active responses >0.01. d, Flow cytometry analysis showing the percentage of IFN-γ⁺ cells in CD4⁺ T cells and CD8⁺ T cells from V_NLRs, V_NHRs and V_R subjects at month 6, after 14h incubation with S^WT, S^D, S^O or S^B. Below: percentages of subjects with active responses >0.01. Data are represented with background subtraction from paired unstimulated controls. The same subjects were analyzed longitudinally at T0 and 6M: V_NLRs (n = 25), V_NHRs (n = 24) and V_R subjects (n = 12). The box plots indicate median, IQR and minimum/maximum. Each dot of the box plots represents the average across three technical replicates of the same subject; the data are pooled from multiple experiments. Statistics were calculated using two-sided Wilcoxon’s signed-rank and two-sided Wilcoxon’s rank-sum test. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001; ^****P < 0.0001.