Phenotypic and functional characterization of HIV-specific CD8⁺ T-cell lines and clones isolated and expanded for infusion. (A) Percentages of HIV-specific CD8⁺ T cells in each cell line (left column) from which each infused clone was ultimately derived. Specific pentamer binding (y-axis) and CD8 (x-axis) are shown. Expression of CD28 (right column) on gated HIV-specific pentamer⁺CD8⁺ T cells (bold line) compared with isotype control (gray line). Inset values represent percentages of CD28⁺CD8⁺ T cells. (B) Data for patient 7705 is shown (HLA B*0702/IPRRIRQGL-HIV-ENV) and is representative of HIV-specific CD8⁺ T-cell clones used for adoptive transfer. Lytic activity (top panel) of the HIV-specific CD8⁺ T-cell clone to autologous B-LCL pulsed with decreasing concentrations of peptide, IFNγ secretion (bottom left panel), and binding to the corresponding MHC-peptide pentamer (bottom right panel) is shown. (C) Expression of CD28 on clonal HIV-specific CD8⁺ T cells infused (bold line) for each patient compared with isotype control (gray line). Inset values represent percentages of CD28⁺CD8⁺ T cells. (D) Expression of CD45RA, CD45RO, CD62L, CCR7, and CD127 (bold line) compared with isotype control (gray line) on a representative CD8⁺ T-cell clone used for infusion (7705). (E) Expression of CD103, CCR9, CD11a, CD11b, CD11c, integrin α4 and integrin β7 (bold line) compared with isotype control (gray line). The bottom right panel shows coexpression of integrin α4 and integrin β7 on the clone (black dots) compared with PBMC (gray dots).