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作 者:陈雷[1] 李晓强[1] 张庆波[1] 袁强[1] 尹淑巧[1] 李涛[1] 左岭[1] 程爱国[1]
机构地区:[1]华北煤炭医学院附属医院
出 处:《中国煤炭工业医学杂志》2000年第5期514-515,共2页Chinese Journal of Coal Industry Medicine
摘 要:目的 观察膀胱癌可溶性抗原诱导细胞毒 T细胞产生 ,为构建膀胱癌疫苗及过继性免疫治疗提供实验基础。方法 提取膀胱癌可溶性抗原 (TSA) ,用 TSA、抗 CD-3单抗、IL -2联合诱导外周血单个核细胞 (PBMC) ,动态观察细胞增殖 ,进行细胞表型分析和细胞因子测定 ;并与 IL-2诱导的 L AK细胞 ,抗 CD-3单抗、IL -2诱导的 CD3-AK细胞进行比较。结果 实验组细胞第 8d增殖7.5 2倍 ,第 12 d增殖 2 8.92倍 ,第 16 d增殖 32 .8倍 ,第 2 0 d增殖 36 .96倍 ;L AK细胞第 12 d增殖 5 .72倍 ,第 2 0 d增殖 0 .82倍 ;CD3-AK细胞第 12 d增殖 2 4.46倍 ,第 2 0 d增殖 2 7.72倍。实验组与 L AK组、CD3-AK组比较相差有显著性 (P<0 .0 5 )。第 8d实验组细胞表型分析 :CD4+ 细胞为 16 % ,CD8+ 细胞为 89%。第 8d培养上清肿瘤坏死因子 (TNF)活性达到高峰 ,对 L 92 9细胞的细胞毒性为85 .0 8%。结论 TSA、抗 CD-3单抗、IL-2等诱导 PBMC,产生了以 CD8+ 为主的细胞毒 T细胞 (CTL ) ,这种细胞体外增殖速度快、增殖倍数高、存活时间长 ,并能分泌肿瘤坏死因子。Objective To observe cytotoxic T lymphocytes induced by soluble antigen of bladder carcinoma for the development of tumor vaccine and the application of adoptive immunotherapy for bladder tumor. Methods Tumor soluble antigen (TSA) was extracted from bladder carcinoma. Peripheral blood mononuclear cells (PBMC) were co stimulated by TSA,IL 2 and anti CD3 monoclonal antibody to develop into cytotoxic T lymphocytes. The proliferation, phenotype and cytokine of the CTL were dynamically analysed and compared with LAK and CD3 AK. Results The proliferation folds of CTL were 7.52, 28.92, 32.8 and 36.96 on the days of 8th,12th,16th and 20th respectively; those of LAK were 5.72 and 0.82 on 12th and 20th, and those of CD3 AK were 24.46 and 27.72 on 12th and 20th respectively. The phenotype of CTL showed 16% CD4 +, 89% CD8 + and the cytotoxicity of the supernatant of CTL against L929 cell was 85.08% on the 8th day. Conclusion Soluble antigen of bladder tumor can induce PBMC into CTL which proliferate more rapidly and produce more TNF.
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