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. 2020 Feb 28;16(2):e1008151. doi: 10.1371/journal.ppat.1008151

Fig 4. IFNα induces expression of unspliced HIV RNA ex vivo.

Fig 4

A: Total CD4+ T cells were isolated from PBMC obtained via leukapheresis from people living with HIV (PLWH) on ART and stimulated in the presence of the HIV integrase inhibitor raltegravir (RAL). After 3 days the cells were harvested, RNA was isolated and reverse transcribed into cDNA, and cell-associated (CA) unspliced (US) and multiply spliced (MS) HIV RNA was measured by qPCR. Additionally, cell culture supernatants were collected for the analysis of supernatant (SN) HIV RNA by qPCR. CA-US (B,E,H,K), CA-MS (C,F,I,L) and SN (D,G,J) HIV RNA is shown following stimulation with DMSO, PMA+PHA or romidepsin (B,C,D) or treated with increasing concentrations of IFNα (E,F,G), anti-CD3/CD28+IL-7+IL-2 (αCD3/CD28) with or without 100 U/mL IFNα, (H,I,J) or treated with 100 U/mL IFNα, IFNβ or IFNω (K,L). Black lines indicate median values and dots represent individual donors (n = 6–7 donors). *p<0.05, **p<0.01, ***p<0.001, ns = not significant, as determined by Wilcoxon matched pairs signed rank test.