Directly Infected Resting CD4+T Cells Can Produce HIV Gag without Spreading Infection in a Model of HIV Latency
Figure 5
Protein expression differences are reflected at the transcript level.
A schematic showing the different HIV RNA species and the primers used to detect them is shown in A. The upper portion depicts the viral genome while the lower portion depicts the various RNA transcripts. For the different RNA species, black boxes represent regions present in all forms of the indicated viral transcript while white boxes represent regions that may or may not be present. GF and GR were used to detect gag levels. RF and ER were used to detect env. RF and VR were used to detect vif, and RF and TR were used to detect tat/rev. Primers were confirmed to solely detect their respective products by gel electrophoresis. Cells were treated and infected as in Figure 4. Cells were cultured in the presence or absence of the integrase inhibitor, raltegravir. RNA was collected at 72 hours (48 hours in CD3/28 treated cells). RNA levels were calculated per cell and were normalized to integration levels. gag levels were obtained by subtracting the levels of gag/cell in the raltegravir treated fraction from the levels in the - raltegravir fraction. An average of 3 experiments in 3 different donors is shown (B). In C, resting cells were infected with or without spinoculation and env and vif levels were measured as in B. An average of 3 experiments in 3 different donors is shown. Error bars represent the standard error of the measurements. *Statistically different at the p<0.05 level.