DC负载肝癌相关抗原后诱导CTL的效应与机理研究

Effect and mechanism of CTL primed by hepatocellular carcinoma associated antigens pulsed dendritic cell

目的 :探讨DC负载肝癌相关抗原后诱导CTL的效应与机理。方法 :用GM -CSF和IL - 4从健康人外周血诱导DC ,使之负载肝癌相关抗原 ,用流式细胞术检测负载肝癌抗原后DC对自身淋巴细胞的活化 ;分别用MTT法和ELISA检测负载肝癌相关抗原后DC活化的淋巴细胞培养上清中TNF和IL - 1 2水平 ;用原位杂交检测淋巴细胞内IFN -γmRNA表达 ;用流式细胞术检测负载肝癌相关抗原DC活化后淋巴细胞表面Fas-L表达 ;最后检测负载肝癌相关抗原DC活化的淋巴细胞对肝癌细胞的特异性杀伤及淋巴细胞培养上清对肝癌细胞的非特异性抑制。结果 :负载肝癌相关抗原DC在体外诱导活化的淋巴细胞在体外对肝癌细胞的杀伤以特异性CTL为主 ,同时分泌较高水平Th1型细胞因子IL - 1 2和TNF ,负载抗原DC活化的淋巴细胞培养上清对肝癌细胞也具非特异性抑制作用。结论

Objective:Get high purity DCs and study the immune responses mechanism of lymphocytes primed by Hepatocellular carcinoma (HCC) associated antigens pulsed DC.Methods:The expression of IFN-γ mRNA in lymphocytes after actived by antigens pulsed DC and the expression of Fas-L on lymphocytes were detected by in situ hybridization and flow cytometry respectively. Specific cytotoxity lysis of T lymphocytes and nonspecific inhibition of liquids in culture medium against HCC lines were also tested. Results:After being stimulated by antigens pulsed DC, the CD56 molecule on lymphocytes decreased . The numbers of CD3 + T / CD4 + T lymphocytes increased more than CD3 + T/ CD8 + T lymphocytes . The ratio of CD4/CD8 changed from 0.84 to 1.04 after activation , but the numbers of γδ T lymphocytes did not change. Primed lymphocytes not only had specific lysis on SMMC-7721 cell line but also had the cytotoxicity against other 3 HCC lines.Conclusion :The results of this research will supply new approach for specific immunotherapy of HCC .