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目的探讨化学交联法构建的Hca-F抗原-卡介苗热休克蛋白70(HSP70BCG)复合物对Hca-F肝癌的主动免疫保护效应及其机制。方法采用ADP亲和层析方法提取HSP70BCG,超速离心法提取粗制肿瘤抗原,采用化学连接法或加入ADP构建Hca-F抗原-HSP70BCG复合物,免疫正常小鼠,3次后检测对Hca-F攻击的主动免疫保化效应,用四甲基偶氮唑蓝(MTT)法检测Hca-F刺激下的脾细胞增殖反应,用流式细胞仪检测脾细胞表型的变化。结果化学交联法和加入ADP构建的Hca-F抗原-HSP70BCG复合物免疫可延长小鼠的存活时间,增强脾细胞对抗原刺激的应答能力和细胞介导的细胞毒活性、提高γδT细胞的百分率。结论 Hca-F抗原-HSP70BCG复合物免疫可诱导抗瘤免疫保护效应。
Objective To investigate the protective effect of Hca-F antigen-BCG HSP70BCG complex on Hca-F hepatocarcinoma induced by chemical cross-linking and its mechanism. Methods HSP70BCG was extracted by ADP affinity chromatography and crude tumor antigens were extracted by ultracentrifugation. Hca-F antigen-HSP70BCG complex was constructed by chemical ligation method or ADP, and normal mice were immunized. After 3 times, Hca-F The active immunopotentiation effect was detected by MTT assay. The proliferative response of spleen cells stimulated by Hca-F was detected by MTT assay. The phenotype of splenocytes was detected by flow cytometry. Results The chemical cross-linking method and Hca-F antigen-HSP70BCG complex constructed by adding ADP could prolong the survival time of mice, enhance the ability of splenocytes to repress antigen stimulation and cell-mediated cytotoxicity, and increase the percentage of γδT cells . Conclusion Immunization with Hca-F antigen-HSP70BCG complex can induce anti-tumor immune protective effect.