Ge H, Bao Z
Shanghai Institute of Immunology, Shanghai Second Medical University.
Zhonghua Yi Xue Za Zhi. 1996 Sep;76(9):658-61.
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.
从人黑色素瘤细胞系中提取与HLA - A2相关的肽段,用于研究HLA - A2限制性肿瘤浸润淋巴细胞(TIL)识别的人黑色素瘤特异性肽抗原。
TIL取自HLA - A2 + 人黑色素瘤患者的外周淋巴结,通过对去污剂溶解的细胞沉淀进行免疫亲和柱层析从黑色素瘤细胞系中纯化HLA - A2分子。与HLA - A2分子结合的肽段经酸洗脱后,通过反相RP - HPLC进行分离。通过将各个级分添加到HLA - A2 + 抗原加工突变细胞T2中,评估其重建黑色素瘤特异性表位的能力。
这些TIL可裂解HLA - A2 + 自体和同种异体黑色素瘤,但不能裂解HLA - A2 - 黑色素瘤。它们也不能裂解HLA - A2 + 非黑色素瘤细胞系。重建级分的RP - HPLC分离显示出三个重建峰(级分)。
这些结果表明,来自三个活性级分的肽段与人黑色素瘤特异性肽抗原相关,这些抗原可被HLA - A2限制性TIL识别。