Fig. 2.

Significant contribution of CCR6⁺ T_CM to the pool of HIV DNA reservoir.

ART, antiretroviral therapy; T_CM, central memory T cell; T_EM/TM, effector T cell. Peripheral blood mononuclear cell from HIV⁺ on ART individuals and uninfected controls (HIV⁻) were stained with a cocktail of CD3, CD4, CD45RA, CCR6, and CCR7 Abs. Shown are (a) the gating strategy for the identification of CCR6⁺ and CCR6⁻ T cells with T_CM (CD45RA⁻CCR7⁺) and T_EM/TM (CD45RA⁻CCR7⁻) cells and (b) the frequency of these subsets in HIV⁻ (n = 13) versus HIV⁺ on ART (n = 20) individuals. Further, CCR6⁺ and CCR6⁻ T cells with T_CM and T_EM/TM phenotypes were sorted by flow cytometry from HIV⁺ on ART individuals and integrated HIV DNA levels were quantified. Shown are (c) the cell purity after sort and (d and e) levels of integrated HIV DNA in subsets isolated from n = 5 HIV⁺ on ART individual (mean ± SEM). (f) The relative contribution of CCR6⁺/CCR6⁻ T_CM and T_EM/TM to the pool of integrated HIV DNA within total memory T cells was calculated considering the frequency of each subset as following: [relative pool of HIV-infected cells per subset (HIV DNA copies per 10e6 cells for each subset × subset frequency/100)/pool of HIV-infected cells per total memory T cells (sum of the relative pool of HIV-infected cells in all four subsets) × 100]. Indicated in the graphs are Mann–Whitney (b) and Friedman and Dunn's multiple comparison test P values (^∗, P < 0.05; ^∗∗, P < 0.01; ^∗∗∗, P < 0.001) (e and f).