T-cell responses targeting HIV Nef uniquely correlate with infected cell frequencies after long-term antiretroviral therapy
Fig 2
Comparing recognition of latently-infected and partially reactivated HIV-infected cells by Nef- and Gag-specific CD8+ T-cells.
A. Timeline for generation of primary cell latency model. B. Characterization of latently-infected cells from two ART-treated participants showing a lack of Gag expressing cells post-sorting on Day 17, and reactivation of Gag expression from these populations following 48 hours of stimulation with anti-CD3/anti-CD28. D. Latently-infected cells from Day 17 (post-sort) were used for CD8+ T-cell recognition assays. Flow cytometry data from 10 hours of stimulation with bryostatin, or unstimulated controls, show a lack of detectable induction of Gag expression at this early time point. CD4 downregulation is observed as a result of bryostatin stimulation. Note that these are the same cells that show Gag expression following 48 hours of stimulation in B. D—F. Cells from this 10 hour stimulation time point were co-cultured with autologous HIV-Gag-specific (D), CMV-pp65-specific (neg control, E), or HIV-Nef-specific (F) CD8+ T-cell clones. Recognition of target cells by CD8+ T-cell clones was measured by degranulation (CD107a exposure). The results show recognition of latently-infected and partially reactivated cells by Nef-specific CD8+ T-cell clones, contemporaneous with a lack of detectable Gag expression and lack of recognition by Gag-specific CD8+ T-cells. Mean ± SD values are shown and p values were calculated by Student’s T test.