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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 the production of cytotoxic T cells by soluble antigen of bladder cancer and provide experimental basis for constructing bladder cancer vaccine and adoptive immunotherapy. Methods TSA, TSA, anti-CD-3 monoclonal antibody and IL-2 were used to induce peripheral blood mononuclear cells (PBMCs). Cell proliferation was measured dynamically. Cell phenotypes and cytokines were measured. Compared with IL-2 induced L AK cells, anti-CD-3 mAb, IL-2 induced CD3-AK cells. Results The cells in the experimental group proliferated 7.52 times on the 8th day, 2992nd on the 12th day, 32.8 times on the 16th day, 36.96 times on the 20th day, The proliferation of CD3-AK cells was 2.46 times on day 12 and 2 7.72 times on day 20. There was a significant difference between experimental group and L AK group and CD3-AK group (P <0.05). On the 8th day, the cell phenotypes of the experiment group were 16% for CD4 + cells and 89% for CD8 + cells. The tumor necrosis factor (TNF) activity peaked on the 8th day, and the cytotoxicity on L 92 9 cells was 85.08%. Conclusion TSA, anti-CD-3 mAb and IL-2 induced PBMCs to produce cytotoxic T lymphocytes (CTLs) that are predominantly CD8 +. These cells proliferated rapidly in vitro with high multiplication rate and long survival time. Secretion of tumor necrosis factor.