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本研究采用酶切等方法将质粒pSVTP1中LMP2a (latentmembraneprotein 2a)的编码基因克隆至pGEM 3Zf+,再经体外转录系统转录获得EBV LMP2aRNA ,将以此RNA激发的DC所诱导生成的CTL作为效应细胞分别与含EBV基因之一EBNA1、EBNA2、EBNA3c、LMP2a的重组病毒感染的正常人成纤维细胞混合后 ,采用LDH释放法检测细胞毒活性。结果显示 ,经体外转录的LMP2aRNA激发的DC所诱导的淋巴细胞对表达LMP2a抗原的成纤维细胞产生特异性的细胞毒活性 ,证实了这种RNA激发的DC能有效地诱导EBV特异性CTL的生成 ,为EBV相关肿瘤的免疫治疗提供了新的实验依据。
In this study, we cloned the encoding gene of LMP2a (latentmembraneprotein 2a) in plasmid pSVTP1 into pGEM3Zf + by restriction enzyme digestion and then transcribed EBV LMP2aRNA by in vitro transcription system. CTLs induced by DC stimulated by this RNA were used as effector cells respectively After being mixed with normal human fibroblasts infected with recombinant virus containing one of the EBV genes, EBNA1, EBNA2, EBNA3c and LMP2a, the cytotoxic activity was examined by LDH release assay. The results showed that lymphocytes induced by DCs stimulated by LMP2aRNA stimulated in vitro produced specific cytotoxic activity on fibroblasts expressing LMP2a antigen and confirmed that this RNA-primed DC effectively induced EBV-specific CTL formation , Provides a new experimental basis for the immunotherapy of EBV-related tumors.