体外培养肿瘤细胞系MHC分型及定量检测

为了对肿瘤细胞进行HLA分型,首先我们应用经肿瘤细胞吸收后的抗血清进行间接淋巴细胞毒试验,对五种人肝癌细胞进行了HLA-A位点常见等位基因A2和A11的检测.结果表明,该方法具有一定的敏感性,对高表达的HLA-A2具有较好的可靠性,但对A11位点检测不明确.使用可获得的抗血清,采用流式细胞免疫荧光法(FACS)进行分析,结果不仅证实了上述结果,而且发现A2和A11分子的表达量普遍低于正常人外周血淋巴细胞.在此基础上,进一步应用该法分析了人肝癌细胞SMMC7721经IFN-γ基因修饰后的HLA-A2表达.

Typing and Quantitative Analysis of MHC on Tumor Cell Lines

In order to type some HLA class I antigens in HCC cell lines, indirect microlymphocytotoxic assay using antiserum absorbed by tumor cells was first carried out. HLA-A2 expression was determined in some HCC cell lines, while HLA-All expression was uncertain. By FACS analysis using the same antisera, we not only confirmed the expression of HLA-A2 and All in these HCC cell lines, but also found that the expression levels of these antigens were much lower than that of peripheral lymphocytes from normal controls. Furthermore, increased HLA-A2 expression was observed on SMMC7721 cells following transduction of IFN-γ gene. The above described techniques may be helpful to establish allogeneic tumor vaccine bank.