Fig. 1
From: Intron-containing RNA from the HIV-1 provirus activates type I interferon and inflammatory cytokines
HIV-1 transduction matures DCs. a Schematic of HIV-1-GFP, with frameshift in env (red line) and gfp in place of nef29, and of the minimal lentivector, with self-inactivating ΔU3 LTR42 and gfp driven by the SFFV promoter16. Unless indicated otherwise, vectors were pseudotyped with VSV G and cells were co-transduced with SIVMAC251 VLPs bearing Vpx. b Flow cytometry of DCs for GFP and CD86, after treatment as indicated. c Flow cytometry histograms for the indicated markers 72 h after DC transduction with HIV-1 (red) or mock (black). d Flow cytometry of DCs for GFP and CD86 after transduction with single-cycle clones, HIV-1NL4-3, HIV-1AD17, HIV-1Z331M-TF, or HIV-1ZM249M. e Transduction of DCs with HIV-2ROD-GFP, single-cycle vector. f Flow cytometry for CD86 and ISG15 of DCs treated for 24 h in the presence of nevirapine with a 1:1000 dilution of supernatant from autologous DCs transduced with the indicated vectors. g DC transduction with HIV-1-GFP in the absence of Vpx and the presence of 2 mM nucleosides. h 12-day spreading infection on DCs, with macrophage-tropic or T cell-tropic, replication-competent HIV-1, with or without SIV VLPs. i qRT-PCR quantitation of CXCL10 (black), IFNB1 (gray), or IL15 (white) mRNAs from DCs transduced with HIV-1-GFP. j qRT-PCR quantitation of CXCL10 mRNA in DCs transduced with either HIV-1-GFP or minimal lentivector, assessed at the indicated times post-transduction. k Cytokines in DC supernatant as assessed by luminex, 72 h after transduction with HIV-1-GFP (black) or minimal lentivector (gray). Shown are blood donor data representative of n = 12 (b), n = 4 (c–k), or n = 8 (h). To determine significance, the MFI of all live cells for each sample was calculated as fold-change versus control. The exception being h where the MFI of only GFP+ cells was compared. When data from each donor replicate within a experiment was combined, the difference in MFI for all experimental vs control conditions was significant in all cases, p < 0.01; one-way ANOVA, Dunnett’s post-test. qRT-PCR and Luminex data were mean ± SD, p < 0.0001; two-way ANOVA, Dunnett’s post-test
