[Human melanoma-specific peptide antigens recognized by HLA-A2 restricted tumor-infiltrating lymphocytes]

HLA-A2-associated peptides were extracted from human melanoma cell lines and used to study human melanoma-specific peptide antigens for HLA-A2-restricted tumor-infiltrating lymphocyte (TIL).

TILS were derived from the peripheral lymph nodes of HLA-A2+ human melanoma patients, HLA-A2 molecules were purified from the melanoma cell lines by immunoaffinity column chromatography of detergent-solubilized cell pellets. Peptides bound to the HLA-A2 molecules were acid eluted and fractionated by reversed phase RP-HPLC. Individual fractions were assessed for their ability to reconstitute melanoma-specific epitopes by adding to the HLA-A2+ Ag-processing mutant cell, T2.

These TIL lysed HLA-A2+ autologous and allogeneic melanomas, but not the HLA-A2- melanomas. They also did not lyse the HLA-A2+ non-melanoma cell lines. The RP-HPLC separations of reconstituting fractions revealed three peaks (fractions) of reconstitution.

These results showed that peptides derived from three active fractions were related to human melanoma-specific peptide antigens recognized by HLA-A2-restricted TIL.