Fig. 5.

ISG expression analysis in a primary CD4 + T cell model of HIV suppression. a Genome organization of the NanoLuc HIV-1 reporter virus. b HIV infection and ART suppression protocol. Primary CD4 + T cells isolated from healthy human PBMC were cultured for 5 days in homeostatic cytokines (IL-2, IL-7, IL-15) then mock-infected (media) or infected with NanoLuc HIV at MOI 2.0 (Donors 1 & 2) or MOI 1.0 (Donor 3). 24 h after infection cells were treated with ART (10 μm raltegravir and 1 μm efavirenz) for 7 days, then stimulated with IFNβ (0, 20, or 100 IU/ml) for 8 h. c Luciferase expression analysis of supernatant from primary CD4 + T cells at indicated times. Three independent experiments were performed. Values represent 12 technical replicates for cells from one donor (#1) in one representative experiment (see Additional file 1: Figure S4B for additional donors). d–f qRT-PCR analyses of mock-infected or HIV-infected CD4 + T cells after IFNβ treatment. Bars represent mean FC ± SD of IFN-stimulated relative to untreated, mock-infected cells. Multiple independent experiments were performed, and data is shown from one representative experiment with three biological replicates per treatment condition. Statistical significance between mock-infected and HIV-infected cells was determined by two-tailed T test with multiple comparisons (Holm-Sidak). *p < 0.05, **p < 0.01, ***p < 0.001