Fig. 7. TLR8 increases HIV replication and reverses latency in CD4+ T cells.

a Viral outgrowth assay: CD4+ T cells isolated from aviremic patients were activated with γ-irradiated PBMCs and PHA (nine donors) or left untreated (five donors) and stimulated with 5 μg/ml of CL264 (TLR7), CL75, or pU/pLA (TLR8) for the time points indicated. Cells from four treated and all five untreated donors were stimulated with SAHA (335 nM), Bryostatin (10 nM), or PMA/ionomycin (500 nM/1.3 μM) for 16 h. HIV RNA was quantified in the supernatant by qPCR. b, c CD4+ T cells isolated from healthy blood donors were TCR-activated in the presence or absence of 5 μg/ml CL264, R848 (TLR7/8), CL75, pU/pLA, or CpG (TLR9, 5 μM) for 48 h prior to infection with cell-free NLENG1-IRES (X4 HIV-1). The frequency of eGFP+ T cells over time was quantified using flow cytometry; representative dot plot examples of untreated and CL75-treated CD4+ T cells 1–5 days post HIV infection are shown in b. Quantification in c is presented as fold change relative to HIV-1-infected cells in the absence of TLR ligands. The mean value of five independent experiment is indicated. Statistical significance was determined using repeated measures mixed effects model with Dunnettʼs post-test on log-transformed data; significance levels: *p < 0.05; **p < 0.01; ***p < 0.001. Source data are provided as a Source Data File.