肿瘤浸润性淋巴细胞识别的人黑色素瘤特异性抗原肽的研究

Human melanoma-specific peptide antigens recognized by HLA-A2 restricted tumor-infiltrating lym-phocytes

从人黑色素瘤细胞中分离人白细胞抗原（ＨＬＡ－Ａ２）相关多肽，并对ＨＬＡ－Ａ２限制性肿瘤浸润性淋巴细胞（ＴＩＬ）识别的黑色素瘤特异性抗原肽进行研究。方法ＴＩＬ来源于ＨＬＡ－Ａ２＋黑色素瘤病人的周围淋巴结。通过去垢剂溶解细胞和亲和层析法从人黑色素瘤细胞中制备ＨＬＡ－Ａ２分子及相关多肽，并经酸处理和反相－高效液相色谱（ＲＰ－ＨＰＬＣ）分离得到不同多肽组份，将其与抗原加工缺陷的ＨＬＡ－Ａ２＋细胞Ｔ２反应，进行重建ＴＩＬ识别的人黑色素瘤特异表位测定。结果ＴＩＬ对自身或同种异体的ＨＬＡ－Ａ２＋的黑色素瘤细胞具有杀伤作用，而对ＨＬＡ－Ａ２－的黑色素瘤细胞和ＨＬＡ－Ａ２＋的非黑色素瘤细胞无作用。从ＲＰ－ＨＰＬＣ分离不同组份的重建试验中，发现有三个活性高峰值。结论具有活性的多肽分子与ＨＬＡ－Ａ２限制性ＴＩＬ识别的人黑色素瘤特异性抗原肽有关。

Objective: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 ) . Methods TILS were derived from the peripheral lympb nodes of HLA-A2+ human melanoma patients , HLA-A2 molecules were purified from the melanoma cell lines by immunoaffinity column chro- matography of detergent-solubilized cell pellets. Peptides bound to the HLA-A2 molecules were acid elut- ed 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 . Re- sults 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. Conclusion These results showed that peptides derived from three active fractions were related to human melanoma-specific pep- tide antigens recognized by HLA-A2-restricted TIL.