Figure 2

Elimination of non-productively infected CD4⁺cells. (A) Representative FACS plots demonstrating the gating scheme employed for the calculation of normalized percent elimination. Cells in culture were stained with anti-CD3 and anti-CD8 antibodies to distinguish targets (CD3⁺/CD8-) and effector (CD3⁺/CD8⁺) populations. Target cells were then gated to determine the percent of gag positive cells, as determined by intracellular staining with an anti-Gag antibody. Uninfected target cells were used as a negative control. (B) An elimination assay was performed to determine the ability of CD8 T cells from B*57/5801⁺ ES (n=10; blue squares), B*57/5801⁺CP (n=9; orange squares), B*57/5801- CP (n=8; red squares) and healthy donors (HD, n=6; purple squares) to reduce the frequency of Gag positive target cells. Unstimulated CD8⁺ T cells or Gag Stimulated CD8⁺ T cells were co-cultured with untreated or EFV treated, autologous CD4⁺ T cell targets at various effector to target ratios. Elimination was analyzed after 6, 18 and 72 hours post infection. Data points where the level of elimination mediated by the ES CD8 T cells was significantly higher than all other experimental groups are indicated (black asterisks, p<.05). (C) The normalized percent elimination for ES Gag-stimulated and unstimulated effectors, for both untreated and 10 μM EFV treated were analyzed at 72 hours post infection for a 1:1 and 1:8 effector to target ratio. For all treatment groups, no statistical difference in the levels of elimination was observed. Median elimination levels are indicated.