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研究目的探讨肿瘤相关抗原的表达及在肿瘤免疫中的作用。研究设计用人Hela细胞株提取含热休克蛋白的ATP结合蛋白,制取单抗,分析相应热休克蛋白在肿瘤细胞株及人体肿瘤中的表达。研究对象 Hela、W14、Brash-2等肿瘤细胞株,WFB、BALB_3T_3正常细胞株,人食管、胃等癌组织。处理方法按Garsia法将提取的ATP结合蛋白免疫BALB/c小鼠,制取杂交瘤,所得单抗对肿瘤细胞株及人体肿瘤作免疫组化及流式细胞分析。结果获取一株稳定分泌IgM的杂交瘤H3F。H3F抗原具有热及化学诱导性,是一种分子量为90kD的热休克蛋白,可在多种肿瘤细胞株及人体肿瘤细胞表面表达,并与浸润性γδT淋巴细胞呈正相关系。H3F在正常细胞株及人体细胞表面阴性。结论 H3F为一种瘤相关抗原,可介异γδT淋巴细胞对肿瘤的免疫监视作用。
Objective To investigate the expression of tumor-associated antigen and its role in tumor immunity. Study and design Human Hela cell line was used to extract heat shock protein-containing ATP binding protein to prepare monoclonal antibody and analyze the expression of the corresponding heat shock protein in tumor cell lines and human tumors. Research targets Hela, W14, Brash-2 tumor cell lines, WFB, BALB_3T_3 normal cell lines, human esophageal, gastric and other cancer tissues. Treatment methods BALB/c mice were immunized with the extracted ATP-binding protein according to the Garcia method to prepare hybridomas. The obtained monoclonal antibodies were used for immunohistochemistry and flow cytometric analysis of tumor cell lines and human tumors. Results A hybridoma H3F stably secreting IgM was obtained. H3F antigen has heat and chemical inducibility and is a heat shock protein with a molecular weight of 90 kD. It can be expressed on a variety of tumor cell lines and on the surface of human tumor cells, and has a positive relationship with invasive γδ T lymphocytes. H3F is negative on the surface of normal cell lines and human cells. Conclusion H3F is a tumor-associated antigen that can mediate the immune surveillance of tumors by γδT lymphocytes.