Increased CD95+ or CD160+ double-negative T-cell subsets are associated with incomplete immune reconstitution in ART-treated people living with HIV

Objective To investigate the expression patterns of CD95 and CD160 on double-negative T (DN T) cells and their association with incomplete immune reconstitution in people living with HIV. Methods In this cross-sectional study, people living with HIV (PLWH) receiving long-term antiretroviral therapy (ART) with sustained viral suppression were classified as immunological non-responders (INR) or immunological responders (IR). Flow cytometry was used to characterize double-negative T (DN T) cells and their CD95 + and CD160 + subsets. Associations between DN T-cell phenotypes and INR were evaluated using univariable and multivariable logistic regression analyses, with odds ratios (ORs) calculated per 10% increase in cell percentages. Receiver operating characteristic (ROC) curves were used to assess discriminatory ability. Results The frequency of DN T cells was significantly higher in the INR group than in the IR group (P = 0.022). The frequencies of CD95 + DN T cells and CD160 + DN T cells were increased in INR participants (P < 0.001 and P = 0.003, respectively) and were negatively correlated with CD4 + T-cell counts (CD95: r = −0.447, P < 0.001; CD160: r = −0.281, P = 0.008). After adjustment for age, ART duration, and baseline CD4 + T-cell counts, multivariable analysis showed that CD95 + DN T cells and CD160 + DN T cells were independently associated with INR. ROC analysis showed moderate discriminatory ability for CD95 + DN T cells (AUC = 0.788), whereas CD160 + DN T cells demonstrated lower discriminatory performance (AUC = 0.687). Adding DN T-cell phenotypic parameters to a baseline clinical model modestly improved discriminatory performance. Conclusion Incomplete immune reconstitution in PLWH is associated with phenotypic alterations of DN T cells characterized by increased CD95 and CD160 expression. These findings provide a phenotypic characterization of DN T-cell alterations associated with immune dysregulation in INR and may offer insights for further mechanistic investigation.