Fig. 8.

Antigen presentation in ascites tumor cells. a HLA-A2 expression by tumor cells. Tumor cell line cells were stained with FITC anti-HLA-A2 or isotype control and analyzed by flow cytometry. b RT-PCR of antigen presentation pathway components. cDNA (RT+) or mock cDNA made in the absence of RT (RT−) from normal donor PBMC, the tumor cell line, or TAP deficient T2 cells was analyzed for expression of TAP1, TAP2, LMP2, LMP7, and LMP10. 1 normal donor RT+, 2 normal donor RT−; 3 tumor cell line RT+, 4 tumor cell line RT−; 5 T2 cells RT+, 6 T2 cells RT−. c Purified CD8⁺ T cells from a normal donor were cultured together with tumor cell line cells that were either not loaded with peptide or pulsed with EBV peptide. The culture was performed in normal medium or in the presence of 50% ascites fluid. The number of IFN-γ spots/10⁵ CD8⁺ T cells plated is shown. d T cell lines specific for Melan-A and gp100 peptides were stimulated with the patient’s tumor cell line and ELISpot analysis was performed. Stimulation with K562-A2 cells is shown as a control