论文部分内容阅读
目的 :研究 HSP70复合物抗人肝癌细胞的免疫效应。方法 :用经 42℃热休克处理的 SMMC- 772 1细胞制备裂解液 ,免疫共沉淀法提取 HPS70复合物 ,采用 SDS- PAGE和 Western- blot鉴定蛋白性质及分子量 ;蛋白测定仪进行复合物定量。 MTT法检测 HSP70复合物体外致敏人脾淋巴细胞对 SMMC- 772 1的细胞毒作用。结果 :SDS- PAGE和 Western- blot证实获得相对分子量约为 7× 10 4的 HSP70蛋白 ;体外细胞毒实施显示在不同效靶比时 ,致敏的淋巴细胞对 SMMC- 772 1均具有一定的杀伤效应 ,且这种效应可被 CD4、CD8的单抗部分阻断。结论 :HSP70复合物能在体外诱导针对人肝癌细胞的淋巴细胞毒效应 ,CD4、CD8介导了部分抗瘤作用 ,提示 HSP70可能通过多种免疫途径参与抗瘤效应。HSP70复合物抗人肝癌免疫的初步研究为今后临床肝癌瘤苗的研制和生物治疗奠定了良好的基础
Objective: To study the immune effect of HSP70 complex against human hepatoma cells. METHODS: The lysates were prepared from SMMC-772 1 cells treated with heat shock at 42°C. HPS70 complexes were extracted by co-immunoprecipitation. Protein properties and molecular weights were identified by SDS-PAGE and Western-blot, and the complexes were quantified by protein analyzer. MTT assay was used to detect the cytotoxicity of externally sensitized human splenic lymphocytes against SMMC-772 1 in the HSP70 complex. RESULTS: SDS-PAGE and Western-blot confirmed that the HSP70 protein with a relative molecular weight of about 7×10 4 was obtained. In vitro cytotoxicity showed that the sensitized lymphocytes had a certain killing effect on SMMC-772 1 at different effect ratios. Effect, and this effect can be partially blocked by the monoclonal antibodies of CD4 and CD8. Conclusion : HSP70 complex can induce cytotoxicity against human hepatoma cells in vitro. CD4 and CD8 mediate part of the antitumor effect, suggesting that HSP70 may participate in the antitumor effect through multiple immune pathways. The preliminary study on the anti-human hepatocellular carcinoma immunity of HSP70 complex laid a good foundation for the future development and biological treatment of clinical hepatocellular carcinoma vaccine.