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Fig. 6 | Retrovirology

Fig. 6

From: Opposing roles of CLK SR kinases in controlling HIV-1 gene expression and latency

Fig. 6

Compound 1H3 inhibits HIV-1 replication in PBMCs, CD4+ T cells, and macrophages. a HIV-1 BaL replication in PBMCs over a period of 8 days post infection as measured by p24 antigen ELISA. PBMCs from healthy human donor were infected with HIV-1 BaL (MOI < 0.01) and treated on day 0 and day 4 post infection with DMSO, AZT (3.74 µM), or 1H3 (0.25 µM). b The effect of increasing concentration of 1H3 on HIV-1 BaL virion production in PBMCs. Indicated doses of the compounds were added following infection, media harvested after 6 days, and virus replication measured by p24 ELISA. Values are expressed relative to p24 levels in DMSO-treated cultures at day 6. The effect of the compounds on cell viability was measured by trypan blue exclusion assays using Glasstic slides (Kova). n = 3. c, d CD4+ T cells from healthy human donors were infected with HIV-1 89.6 then treated immediately with DMSO, 10 µM TG003, or 300 nM 1H3 for 3 days. Cells were harvested and the effect of individual treatments on HIV-1 c protein or d RNA accumulation assessed by western blot and RT-qPCR, respectively. Shown are results from 6 independent experiments with 8–10 donors. e Effect of 1H3 on HIV-1 replication in human macrophages. Monocyte-derived macrophages from healthy donors were infected for 2 days with HIV-1 89.6, then treated with DMSO, a combined antiretroviral drug Lamivudine/Lopinavir + Ritonavir (1.5 µM/53 nM) or 1H3 (300 nM). After 10 days, cells were fixed, stained to detect intracellular levels of Gag and analyzed by flow cytometry to assess changes in %Gag positive cells and Gag mean fluorescence intensity (MFI). Shown on the left are representative flow cytometry plots of uninfected and infected macrophages and histograms showing shifts in the Gag Mean fluorescence Intensity (MFI) with treatment. On the right, summary data shown are expressed relative to DMSO treated samples (n = 8 from 5 independent experiments). Statistical analysis, one sample t test, ns = not significant, *p < 0.05, **p < 0.01, and ****p < 0.0001

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