Directly Infected Resting CD4+T Cells Can Produce HIV Gag without Spreading Infection in a Model of HIV Latency
Figure 2
Infected resting CD4+T cells can produce HIV Gag.
Bulk unstimulated CD4+T cells were spinoculated and cultured in the presence of the protease inhibitor, saquinavir. Gag protein was measured in the endogenously activated (HLA-DR+, CD25+, or CD69+) and resting (HLA-DR−,CD25−,and CD69−) cells based on activation marker expression at 0, 24, 48, 72, and 96 hours post infection. Control cells were treated with the reverse transcriptase (RT) inhibitor, efavirenz, to establish background protein levels. An average of 3 experiments in 3 different donors is shown (A). In B, purified resting or CCL19 treated cells were infected with HIV with or without spinoculation at 1200×g. Controls and gates were made as in A. Cells were gated on the activation marker negative (HLA-DR−,CD25−,CD69−) population. Approximately 10,000 events were collected for spinoculated samples while approximately 100,000 events were collected for cells that were not spinoculated. Data is a representative of 2 experiments in 2 different donors.