摘要
目的 探讨肝癌DC疫苗诱导肝癌特异性淋巴细胞的效应与机理。方法 用流式细胞术检测肝癌细胞表面HLA-DR的表达;用CM-CSP和IL-4从健康人外周血诱导DC,使之负载肝癌相关抗原,并在BCG HSP-70作用下活化,制成肝癌DC疫苗;用流式细胞术检测BCG HSP 70活化的负载肝癌抗原DC对自身淋巴细胞的活化;分别用MIT法和ELISA检测肝癌DC疫苗活化的淋巴细胞培养上清中TNF和IL-12水平;用原位杂交检测淋巴细胞内IFN-γmRNA表达;用流式细胞术检测经DC疫苗活化后淋巴细胞表面Fas-L表达;最后检测肝癌DC疫苗活化的淋巴细胞对肝癌细胞的特异性杀伤。结果 肝癌DC疫苗在体外诱导活化的淋巴细胞对肝癌细胞的杀伤以CD4^+ T CTL为主,不但可以杀伤SMMC-7721细胞,同时对其它3种肝癌细胞也有不同程度的杀伤。CTL培养上清液和细胞中可分别检测到T_(HI)型细胞因子TNF和IFN-γ mRNA。Fas和Fas配体途径发挥了重要的作用。结论 为肝癌DC疫苗的进一步深入研究及今后的临床应用奠定了良好的基础。
Objective To investigate the effect and mechanism of CTL primed by dendritic cells based Hepatocellular carcinoma vaccine. Methods Eexpression of HLA-DR molecule on surface of HCC lines was analysed. Get high purity DCs and study the immune responses of lymphocytes primed by BCG HSP70 activated DCs which loaded with hepatocellular carcinoma (HC) associated antigens which named DC based hepatocellular carcinoma vaccine. The IFN-γ mRNA in lymphocytes after activated by DC vaccine and the Fas-L expression on DC vaccine primed lymphocytes were detected by in situ hybridization and flow cytometry respectively. Specific cytotoxity lysis of T lymphocytes against HCC lines were also tested. Results After being stimulated by DC based HC vaccine, 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 1.1 after activation. High level TNF and IFN-γ mRNA were also detected in the culture medium of CTL and primed lymphocytes respectively. Primed lymphocytes not only specific lysis SMMC-7721 cell line but also had cytotoxicity against other 3 HCC lines. FAS/FAS-L path way plays an important role in the cytotoxicity of CTL. Conclusion The results of this research may afford new approach for specific immunotherapy of HC.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2003年第5期368-371,374,共5页
Immunological Journal
