Intra- and extra-cellular environments contribute to the fate of HIV-1 infection

Abstract

Highlights d HI.fate reports HIV-1 fate in target cells by using sensitive fluorescent proteins d Alternative fates of HIV-1 infection are linked to differential gene expression d Small molecules can modulate the fate of HIV-1 infection d Extra-cellular environment contributes to the fate of HIV-1 in vivo

T cell migration potentiates HIV infection by enhancing viral fusion and integration

“…The addition of chemokines CCL19 and CCL21, both expressed in the T cell zone of lymph nodes, did not further enhance T cell infection rates (Figure S2). Similarly, infection in collagen matrix did not promote latent HIV infections using the dual-fluorescent protein HIV reporter strain (HI.Fate; Ratnapriya et al, 2021;Figure S2). Together, these data collectively show that T cells embedded in collagen fibrillar networks were more susceptible to productive HIV infection.…”

“…Seemingly random CD4 T cell motility patterns in lymph nodes are in fact regulated by both cell-intrinsic and environment-specific cues that coordinate their search efforts to rapidly detect and eliminate pathogens (Mandl et al, 2012). A recent study illustrates this point, where extracellular cues in various organs differentially modulate HIV replication and latency generation in humanized BLT mice in vivo (Ratnapriya et al, 2021). Together, our reductionist approach to model initial HIV-T cell interactions within a dynamic, fibrillar network suggests that migrating T cells in tissues may be more susceptible to infection than those studied ex vivo under typical suspension cultures.…”

“…The R5-tropic HIV Gag-iGFP reporter was constructed as previously described (Koh et al, 2020a). The R5-tropic HIV-GFP and HIV-GFPDnef viruses were previously reported (Usmani et al, 2019), and the HI-Fate reporter was constructed and produced as described (Ratnapriya et al, 2021). All plasmids were isolated using GenElute HP Plasmid Maxiprep Kit (Sigma) and confirmed by Sanger sequencing.…”

“…Nef-deleted HIV-GFP was generated as previously reported (Usmani et al, 2019). The HI-Fate dual fluorescent reporter (Ratnapriya et al, 2021) contains an LTR promoter that drives expression of E2-Crimson (replacing the Nef locus) and a mammalian Elongation Factor 1a (EF1a)-HTLV chimeric promoter driving expression of ZsGreen1 (schematic diagram in Figure S2. HI-Fate was pseudotyped with VSVg and used for infection studies.…”

Single-Cell Profiling of Latently SIV-Infected CD4 + T Cells Directly Ex Vivo to Reveal Host Factors Supporting Reservoir Persistence

“…The methods used to date to profile infected cells vary and generally fall into three categories: (i) in vitro infection models to generate latently infected cells for characterization ( 33 – 37 ), (ii) ex vivo purification of cells expressing a specific marker hypothesized to enrich for infected cells (e.g., memory cells) followed by viral quantitation in the target population ( 20 , 24 – 26 , 38 , 39 ), and (iii) in vitro reactivation of latently infected cells, followed by detection of viral RNA or protein expression for detailed characterization by flow cytometry or gene expression ( 40 – 43 ). There are limitations to these approaches.…”