ADAR1 deficiency restricts HIV-1 replication in activated human CD4+ T cells.

<p>(a-b) PBMCs from healthy donors (n = 4) and ADAR1-mutated patients (n = 3) were challenged with equivalent infectious units of HIV-1 GFP virus and analyzed on day 5 after infection. The reverse transcriptase inhibitor (RTI) cocktail (10μM AZT, 20μM DDI, and 10μM 3TC) was used as specificity control. (a) Representative dot plots of flow cytometry analysis from PBMCS of an AGS patient and a healthy control. The viable cells (gate, left graphs) were stained and selected for CD3+CD4+ expression (b). The percentages of infected (GFP+) CD4+ T cells (c) are shown in the upper right quadrant. (d) Bar graph representing the mean±s.e.m. of GFP+ infected cells (n = 3), ***P<0.001. (e) Bars represent luciferase activity in relative luciferase units (RLU) of cells infected with a VSV-G virus. The activity was measured in non-stimulated (NS), IL-2-stimulated or RTI-treated cells. Bars represent mean±s.e.m (n = 3). (f) Bar graphs represent mean±s.d (n = 3) of HIV-1 p24 protein amounts released into the cell culture supernatant on day 5 after infection, *P<0.05. Relative quantification of viral DNA pol (g) and viral pol mRNA (h) expressed as fold versus NS control cells. (i) Relative quantification of ISG15, IFIT27, RSAD2, IFIT44L, and OAS1 from HIV-1-infected PBMCs from AGS patients (n = 3) and healthy donors (n = 3) presented as fold over NS control expression.