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目的研究热休克蛋白抗原肽致敏的脐血树突状细胞体外对肺癌细胞的杀伤作用。方法用体外构建的热休克蛋白-抗原肽复合物刺激经组合细胞因子诱导的脐血树突状细胞,MTT法检测其对肿瘤细胞的杀伤作用。结果所得蛋白经电泳及Western blot进行蛋白分子量及性质鉴定为热休克蛋70。热休克蛋白-抗原肽负载可以促进脐血树突状细胞对T细胞的激发作用,使其对靶细胞有了更强的生长抑制作用。各组T细胞杀伤活性分别为:负载抗原组(85.77±1.03)%(正常T细胞)、(45.01±1.66)%(肺癌患者T细胞);未负载抗原组(41.92±1.38)%(正常T细胞)、(13.99±3.07)%(肺癌患者T细胞)。组间比较差异有统计学意义(P<0.01)。结论热休克蛋白抗原肽负载脐血树突状细胞能有效激发外周血T淋巴细胞,使其对靶细胞有了更强的生长抑制作用。本研究为解决树突状细胞的来源及开发特异性CTL的治疗开创了条件。
Objective To study the killing effects of heat shock protein antigen-pulsed umbilical cord blood dendritic cells on lung cancer cells in vitro. Methods Dendritic cells derived from combined umbilical cord blood were stimulated with heat shock protein-antigen peptide complexes constructed in vitro, and their cytotoxic effects on tumor cells were detected by MTT assay. Results The protein was analyzed by electrophoresis and Western blot for the protein molecular weight and properties of heat shocked egg 70. The heat shock protein-antigen peptide load can promote the excitotoxicity of umbilical cord blood dendritic cells to T cells, which makes them have a stronger growth inhibitory effect on the target cells. The killing activity of T cells in each group was (85.77±1.03)% (normal T cells), (45.01±1.66)% (T cells in lung cancer patients), and (41.92±1.38)% (41.92±1.38)% in antigen-free groups (normal T cells). Cell), (13.99±3.07)% (lung cancer patient T cells). There was a statistically significant difference between groups (P<0.01). Conclusion Heat shock protein antigen peptide loaded umbilical cord blood dendritic cells can effectively stimulate peripheral blood T lymphocytes, so that it has a stronger growth inhibitory effect on target cells. This study opened the way for the solution of dendritic cells and the development of specific CTL treatments.