Citation
- Authors: Lopez P. et al.
- Year: 2022
- Journal: Cell Rep 38 110406
- Applications: in vitro / DNA / jetOPTIMUS
- Cell type: HEK-293T
Description: Human embryonic kidney Fibroblast
Known as: HEK293T, 293T
Method
All HIV-1 stocks were generated by transient transfection of HEK 293T cells using JetOptimus reagent. To generate HIV-Gag-iGFP virus, cells were co-transfected with the pGagPol plasmid (NIH HIV Reagents Program). Viral supernatants were collected 48 h after transfection and layered over a 20% sucrose solution before ultra-centrifugation at 35,000 rpm for 90 min using an SW70Ti rotor (Beckman Coulter). Virus pellets were resuspended in PBS, aliquoted and stored at −80°C until use. Each HIV stock was titrated using MAGI.CCR5 cells and expressed as infectious blue focus units (bfu) per ml.
Abstract
T cells actively migrate along reticular networks within lymphoid organs in search for cognate antigen, but how these behaviors impact HIV entry and infection is unclear. Here, we show that migratory T cells in 3D collagen matrix display significantly enhanced infection and integration by cell-free R5-tropic lab adapted and transmitted/founder molecular HIV clones in the absence of exogenous cytokines or cationic polymers. Using two different collagen matrices that either support or restrict T cell migration, we observe high levels of HIV fusion in migratory T cells, whereas non-motile T cells display low viral entry and integration. Motile T cells were less sensitive to combination antiretroviral drugs and were able to freely migrate into regions with high HIV densities, resulting in high infection rates. Together, our studies indicate that the environmental context in which initial HIV-T cell encounters occur modulates HIV-1 entry and integration efficiencies.