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从黑色素瘤病人的瘤块和淋巴结中制备黑色素瘤细胞和肿瘤浸润性淋巴细胞（ＴＩＬ）系，通过流式细胞仪对黑色素瘤细胞表型分析。经去垢剂处理、免疫沉淀和亲和层析法从ＨＬＡＡ２阳性患者黑色素瘤细胞中分离纯化ＨＬＡＡ２蛋白，其纯度通过４％～２０％梯度ＳＤＳＰＡＧＥ和Ｗｅｓｔｅｍｂｌｏｔ鉴定。１００℃沸水浴和弱酸处理ＨＬＡＡ２蛋白经反相高压液相色谱层析（ＲＰＨＰＬＣ）得到不同组份多肽，将其与抗原加工缺陷的ＨＬＡＡ２阳性Ｔ２细胞反应，进行重建瘤细胞特异性表位测定。ＴＩＬ杀伤活性结果表明，来源于６２４黑色素瘤细胞的不同组份多肽中有三个活性峰值，而来源于Ｃｈａｐ黑色素瘤细胞的不同组份多肽中仅一个与上述细胞组份位置相同的单一活性峰值。提示具有活性组份的多肽与ＨＬＡＡ２限制性ＴＩＬ识别的人黑色素瘤细胞抗原肽有关。
Melanoma cells and tumor infiltrating lymphocyte (TIL) lines were prepared from tumor masses and lymph nodes of melanoma patients and melanoma cell phenotypes were analyzed by flow cytometry. The HLAA2 protein was isolated and purified from HLAA2 positive melanoma cells by detergent treatment, immunoprecipitation and affinity chromatography. The purity of the HLAA2 protein was determined by SDS PAGE and Westemblot gradients of 4% to 20%. 100 °C boiling water bath and weak acid treatment of HLA A2 protein by reversed phase high pressure liquid chromatography (RP-HPLC) to obtain different components of the polypeptide, and antigen processing defects in HLA-A2 positive T2 cells, reconstruction of tumor Cell-specific epitope assays. The results of TIL killing activity showed that there were three activity peaks in the polypeptides of different components derived from 624 melanoma cells, and only one of the different component polypeptides derived from Chap melanoma cells had the same position as that of the above-mentioned cell components. Peak. It is suggested that the polypeptide with active component is related to the human melanoma cell antigen peptide recognized by HLA-A2-restricted TIL.