Figure 4.
Different recognition of transporter associated with antigen processing (TAP)-deficient CMT.64 cells by T cells generated by CMT.TAP1/B7.1 cells and by CMT.TAP1/B7.1 cell-lysate-loaded dendritic cells (DCs). Standard 51Cr-release assays were performed using major histocompatibility complex class I (MHC-I) mismatched Mid-T2 (negative control), CMT.64/pp and CMT.TAP1,2 cl.21 cells as targets. (a) Splenocyte-derived T cells were generated as follows. Bone marrow-derived DCs were loaded with lysates of γ-irradiated CMT.TAP1/B7.1 cells for 6 hr, followed by addition of lipopolysaccharide (LPS) for DC maturation. After washing, DCs were injected intraperitoneally (i.p.) into C57BL/6 mice (1 × 106 cells per mouse) for immunization. After a 7-day immunization, the splenocytes were collected and re-stimulated with CMT.TAP1/B7.1 cell-lysate-loaded DCs for 5 days. (b) Splenocyte-derived T cells were generated by immunization of mice with γ-irradiated CMT.TAP1/B7.1 cells (5 × 106 cells/mouse). After a 7-day immunization, the splenocytes were re-stimulated in vitro with γ-irradiated and mitomycin-c-treated CMT.TAP1/B7.1 cells for 4–5 days. One of three experiments is shown.