抗热休克蛋白抗体H3F识别分子的分析及意义

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。

ANALYSIS AND SIGNIFICANCE OF HEAT SHOCK PROTEIN RECOGNIZED BY CORRESPONDING MONOCLONAL ANTIBODY-H3F

Objective To search for the expression of tumor-associated antigen and its function in tumor immunity. Design Extracting ATP—binding protein which containing heat shock protein from Hela cell Line, we developed monoclonal antibody to analyse corresponding heat shock protein expression on the tumor cell lines and human tumors. Objects Hela, W14, Brash-2 etc tumor cell lines. WFB, BALB-3T3 normal cell lines.Human esophageal cancer, gastric cancer and others. Interventions By immunizing BALB/C mouse with ATP-binding protein based on Garsia's method, hybridoma was developed. Using obtained monoclonal antibody. immunohistochemical and flow cytometric analysis were performed on tumor cell lines and human tumors tissues. Results A hybridoma—H3F line which secretes steady IgM was obtained.Heat shock and chemical treatment can induce H3F antigen which is a 90kDa heat shock protein expressing on many tumor cell lines and human tumors, and positively associated with infiltration of gamma-delt T lymphocytes, and negative on normal cell lines and human cells. Conclusion H3F is a tumor-associated antigen which can induce immune surveillance if gamma—delt T lymphocytes to tumors.