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目的 探讨转染原发性肝癌(HCC)mRNA的树突状细胞(DC)能否诱导抗肿瘤特异性细胞毒性T淋巴细胞(CTL)。方法 采用HCC患者外周血单核细胞(PBMC)体外刺激分化为DC细胞;从人肝癌HepG 2细胞和3例HCC患者的肝癌组织中体外扩增mRNA。以mRNA转染DC细胞,并与PBMC混合培养诱导扩增CTL。流式细胞计数仪检测培养细胞中CD3 +、CD4+、CD8+细胞的比例。51Cr释放法测定CTL的杀瘤活性。结果 经扩增人肝癌HepG 2mRNA和2例AFP(+ )患者的AFP(+ )HCCmRNA诱导3周后,CD3 +、CD8+细胞占淋巴细胞总数由诱导前的2 7.8%、2 6.5 %、2 9.6%升高至89.3 %、73 .6%、86.8% ;而经扩增AFP(-)HCCmRNA诱导3周后,CD3 +、CD8+细胞占淋巴细胞总数由诱导前的2 5 .4%升高至5 3 .6%。转染HepG 2细胞和AFP(+ )的患者HCCmRNA的DC诱导的CTL对HepG 2细胞杀瘤活性明显高于AFP(-)的患者,其杀瘤特性由MHC I限制的CD8+T细胞所介导。结论 HCCmRNA体外转染DC能诱导肿瘤特异性CTL ,可为肝癌的免疫治疗提供新的有效手段
Objective To investigate whether dendritic cells (DCs) transfected with primary hepatocellular carcinoma (HCC) mRNA can induce antitumor specific cytotoxic T lymphocytes (CTLs). Methods Peripheral blood mononuclear cells (PBMCs) from patients with HCC were stimulated to differentiate into DCs in vitro. MRNA was amplified from human hepatocellular carcinoma HepG 2 cells and HCCs from 3 patients with HCC in vitro. DCs were transfected with mRNA and mixed with PBMC to induce CTL amplification. The proportion of CD3 +, CD4 +, and CD8 + cells in cultured cells was detected by flow cytometry. 51Cr release assay CTL tumor killing activity. Results After 3 weeks of induction of AFP (+) HCC mRNA in HepG 2 and 2 AFP (+) patients, the total number of CD3 + and CD8 + cells in lymphocytes increased from 2 7.8%, 2 6.5% and 2 9.6 % Increased to 89.3%, 73.6% and 86.8% respectively. After 3 weeks of induction of AFP (-) HCC mRNA, the total number of CD3 + and CD8 + cells in lymphocytes increased from 25.4% before induction 5 3 .6%. DCs induced by DCs induced by HCC mRNA in HepG2 cells and AFP (+) -treated mice showed significantly higher cytotoxicity against HepG2 cells than AFP (-), and their cytotoxicity was mediated by MHC I-restricted CD8 + T cells guide. Conclusion Transfection of DC with HCC mRNA induces tumor-specific CTL in vitro, which may provide a new effective method for immunotherapy of HCC