Figure 3.

Responses of rectosigmoid CD8⁺ T_RM cells to antigenic stimulation. (a) Representative flow cytometry plot demonstrating degranulation (CD107a) and IFN-γ-production following stimulation with an HIV-1 Gag-peptide pool or medium containing DMSO (negative control) by rectosigmoid CD8⁺ T_RM (top) and rT_EFF (bottom) from an HIV-1⁺ controller. (b) Difference in the frequency of CD8⁺ T_RM and rT_EFF cells responding to Gag-peptide and SEB stimulation. (c) Total percentage of rectosigmoid CD8+ T_RM and rT_EFF responding in any way (TNF-α, IFN-γ, IL-2, CD107a, and/or MIP-1β) to HIV-1 Gag stimulation. For each subject, the total HIV-1 Gag-specific response shown in (c) was calculated by combining all non-overlapping Boolean categories that are displayed individually in panels (d), for T_RM, and (e), or rT_EFF. (d) Difference in percentages of poly- and monofunctional Gag-specific rectosigmoid CD8⁺ T_RM cells across HIV-1 disease status. (e) Difference in percentages of poly-and monofunctional Gag-specific rectosigmoid CD8⁺ rT_EFF cells across HIV disease status. Response data were analyzed using Boolean gating of responding populations within live, CD3⁺, CD8⁺, CD45RO⁺ or CD45RO^Neg, CD103⁺CD69⁺S1PR1⁻ gates. Co-expression analyses were generated using SPICE software. Poly- and monofunctional combinations with negligible responses were omitted. Horizontal bars represent medians, whiskers indicate interquartile ranges with the level of significance shown with asterisks as follows: * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001.