食管癌可溶性抗原和超抗原构建肿瘤疫苗的免疫活性成分分析

目的对食管癌可溶性抗原和超抗原SEC构建的肿瘤疫苗中的免疫活性成分进行分析,从而为探讨其抗癌机制打下理论基础。方法提取食管癌抗原,和超抗原SEC构建成肿瘤疫苗;分离人外周血单个核细胞（peripheral blood mononuclear cells,PBMC）,经肿瘤疫苗联合作用,进行体外培养,观察其增殖活性;流式细胞术FCM和细胞毒试验测定效应细胞表型和杀伤活性;提取食管癌细胞中的细胞膜抗原（mAg）、热休克蛋白70（HSP70）、DNA、RNA和细胞内多肽（Peptides）,用于诱导PBMC增殖,培养6d,用3H-Tdr掺入法测定细胞增值,确定其有效成分。结果经肿瘤疫苗刺激的PBMC组增殖活性最强,于72h达到高峰,并且特异性刺激CD8＋T细胞群增值。肿瘤疫苗刺激的PBMC组诱导的CTLs对靶细胞杀伤活性显著高于单纯PBMC组（P〈0.01）。食管癌细胞中的膜抗原和细胞内多肽能显著刺激PBMC增殖。结论食管癌抗原与超抗原构建的肿瘤疫苗能诱导效应细胞明显增殖、活化、并产生高效、特异的抗肿瘤效果,具有免疫活性的成分是细胞中的mAg和细胞内多肽。

Analysis for the immune active component of tumor vaccines established by esophagus cancer soluble antigen and superantigen

Objective To analyze the immune active component of tumor vaccines established by esophagus cancer soluble antigen and superantigen so as to provide theoretic basis for the researches about its anticancer mechanism. Methods Tumor soluble antigens were extracted from esophagus cancer cells and constructed with superantigen SEC into tumor vaccine. Human peripheral blood mononuclear cells （PBMC） were isolated, then stimulated with tumor vaccine and cultured in vitro. The proliferation activity was observed. The phenotypes of effector cells were detected by flow cytometry （FCM） and killing activity was tested by cytotoxic assay. The mAg, heat shock protein70 （HSP70）, DNA, RNA, and intra-cellular polypeptides were extracted from esophagus cancer cells to induce the proliferation of PBMC, and the cells were cultured for 6 days, at last, the cell proliferation activity was detected by3H-Tdr incorporation method to identify its active component. Results The proliferation activity of PBMC stimulated by tumor vaccine was the highest, which reached peak at 72h, furthermore,which specially stimulated the proliferation of CD8 ＋ T cells population. The killing activity of CTLs induced by PBMC stimulated by tumor vaccine was significantly higher than that of simple PBMC group （ P 〈 0.01 ）. Conclusion The tumor antigen constructed by esophagus cancer antigen and superantigen SEC can induce PBMC to activate and proliferate into CD8 ＋ CTL with specific cytotoxicity against the tumor in which mAg and intra-cellular polypeptides possess immune active component.