Figure 8.

iCD4+ T cells have increased resistance to HIV infection. (A) HIV infection challenge in iCD4+ T cells, experimental outline. (B) Rate of HIV-1 or HIV-2 infection, 48 h after infection of CD4⁺ T cells transduced with control (GFP), IRF3, and RELA K5R lentivectors. Cells were transduced, pretreated with 5AZA (2 µM) for 48 h, and subsequently infected with HIV-1 or HIV-2 single-round virus (n = 4 donors combined from two independent experiments). Each symbol represents one donor, bars represent mean ± SEM of four donors, paired one-way ANOVA with Tukey’s multiple comparison test on the highest dose of virus. (C) Pearson correlation of infection rates with IFNλ1 concentration of CD4⁺ T cells transduced and treated with 5AZA as indicated. (D) Western blot of IRF3, IRF7, RELA, MX1, and Actin in CD4⁺ T cells transduced with either GFP or RELA K5R and IRF3, and pretreated with 5AZA (2 µM), control or B18R supernatant for 3 d (representative of two independent experiments). (E) Rate of HIV-1 or HIV-2 infection, 48 h after infection of CD4⁺ T cells transduced with control (GFP), IRF3, and RELA K5R lentivectors. Cells were transduced, pretreated with 5AZA (2 µM), control, or B18R supernatants for 48 h and subsequently infected with HIV-1 or HIV-2 single-round virus (n = 4 donors combined from two independent experiments). Each symbol represents one donor, bars represent mean ± SEM of four donors, paired one-way ANOVA with Tukey’s multiple comparison test on the highest dose of virus. (F) Working model. RELA functions as a rheostat to control IFN-I/III expression levels in CD4⁺ T cells. IFN-I/III expression requires tonic cGAS activity or PRR stimulation and positive feedback from IRF7 signaling. *P ≤ 0.05, **P ≤0.01, ***P ≤ 0.001, ****P ≤ 0.0001. Figure was generated using Biorender. Source data are available for this figure: SourceData F8.