Figure 5.
HLA Class II-Restricted CD8+ T Cells Lyse Autologous HIV-Infected Targets Ex Vivo and Exert Putative Immune Selection Pressure In Vivo
(A) Summary data assessing specific lysis of target cells by class II-restricted CD8+ T cell clones from subjects 474723 and 270245 in a standard 6 hr chromium release assay. Autologous targets included EBV-transformed BCL pulsed with peptide or no peptide, CD4+ T cells infected with the HIV strain NL4.3, and monocyte-derived macrophages infected with VSV-G pseudotyped HIV NL4.3 encoding GFP, all at an effector to target cell ratio (E:T) of 1:1. Statistical significance was determined using a paired t test.
(B) Summary data assessing specific lysis of target cells by ex vivo tetramer+ sorted HLA class I- and class II-restricted CD8+ T cells in a standard 6 hr chromium release assay at multiple E:T ratios. Ex vivo effector cells were derived from fresh blood and tetramer sorted within 12 hr of phlebotomy. As target cells, autologous EBV-transformed BCL from subjects 474723 and 270245 were pulsed with cognate peptide or no peptide. Statistical significance was determined using a paired t test.
(C) Summary data assessing specific lysis of autologous monocyte-derived macrophages pulsed with the cognate peptide Gag41 or infected with VSV-G pseudotyped HIV NL4.3 encoding GFP by ex vivo DR11-Gag41 tetramer+ sorted CD8+ T cells from subject 474723 in a standard 6 hr chromium release assay at an E:T ratio of 1:1.
(D) Deep sequencing of autologous virus in subject 474723. Mutation Q308H within the Gag41 epitope is boxed. Specific lysis of EBV-transformed BCL pulsed with wild-type Gag41 peptide or the Q308H mutant peptide was tested in a standard 6 hr chromium release assay with ex vivo tetramer-sorted DR11-Gag41 tetramer+ sorted CD8+ T cells or bulk CD8+ T cells at a 1:1 E:T ratio.
All data points for each graph represent biological replicates in a single experiment. Error bars represent SD. See also Figure S6.