Antigen-specific retention of CD20-specific cTCR⁺ T cells depends on CD3ζ signaling and LFA-1. (A) Murine CD8⁺ T cells expressing the MB20-18⁺ cTCR exhibit antigen-specific cytolytic activity toward mCD20 expressing cell lines (A20, EL4-CD20), but not control cells (EL4-hCD20, BM185). Standard 5-hour ⁵¹Cr release assay analysis of MB20-18⁺cTCR⁺CD8⁺ T-cell lytic activity. Data are representative of 2 experiments. (B) Calculation of lung to non-lung bioluminescent signal ratio from mice injected intravenously with 10⁷ CBR⁺cTCR⁺Thy1.1⁺CD8⁺ T cells. Data from 2 experiments were overlaid comprising 7 to 8 mice per time point. (C) Lytic activity of cTCR⁻ or cTCR⁺ T cells toward WT B cells and BM185-mCD20. T cells were incubated with targets for 3 hours, and target lysis was determined by a 7AAD assay as described in “In vitro T-cell analysis.” Data are representative of 2 experiments. (D) cTCR⁻ or cTCR⁺ T cells were conjugated with freshly isolated WT B cells, and the percentage of T cells in conjugates was determined by flow cytometry as described in “In vitro T-cell analysis.” Data are representative of 3 experiments. (E) Bioluminescent image of mice 24 hours after receiving 10⁷ CBR⁺MB20-18⁺CD8⁺ T cells that had been treated with anti–LFA-1 or PBS before injection. (F) Serial quantitation of T-cell bioluminescent signal from mice injected with CBR⁺cTCR⁺Thy1.1⁺CD8⁺ T cells. Data are representative of 2 experiments with 3 to 4 mice per group. Bar graphs and data points represent mean ± SE. Bioluminescent images were assembled with the use of Illustrator (Adobe Systems).