Functional characteristics of polyclonal and virus-specific effector CD4+ T cell responses in HIV-infected LNs and blood.

(A) Flow cytometry plots (HIV ART+ subject) and plots for matched HIV-Gag or–Env-specific CD4+ T cell responses in HIV-infected LN and PB. (B) Flow cytometry plots (HIV-infected CP) showing the negative control (NC) and Gag-specific response of LN CD4+ T cell response. The high abundance of CD107a (red) that is not co-expressed with IFNγ or TNF is illustrated in this example. SPICE analysis of functional combination between LN (red) and PB (red-gray) Gag-specific CD4+ T cell responses for HIV-infected CPs and ART+ subjects. (C) Flow plots (HIV-infected CP) of Gag-specific CD4+ T cell response (red) from LN and PB in relation to CD27 and perforin expression. Graphs represent the frequency of (C) perforin+ and (D) T-bet^hi cells between LN and PB Gag-specific CD4+ T cells. (E) Flow plots (HIV-infected CP) of MIP-1α versus IFNγ and TNF production for LN and PB SEB stimulated CD4+ T cells. Corresponding plots showing the frequency of MIP-1α+ SEB stimulated CD4+ T cells (top) and MIP-1α+ of IFNγ/TNF/CD107a/MIP-1α+ SEB stimulated CD4+ T cells (bottom). (F) Flow plots (HIV-infected CP) of MIP-1α versus IFNγ and TNF production for LN and PB CMV-specific CD4+ T cells and corresponding graphs showing the frequency of MIP-1α+ CMV-specific CD4+ T cells (top) and MIP-1α+ of IFNγ/TNF/CD107a/MIP-1α+ CMV-specific CD4+ T cells (bottom). Median and IQR are shown for all bar plots. Permutation test was performed between the pie charts. Wilcoxon matched-pairs single rank tests were performed to compare differences between two matched groups; *P < 0.05, **P < 0.01 and ***P < 0.001. All data-points are derived from the Mexico cohort.