摘要
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 。
Objective To induce efficient expansion of cytotoxic T lymphocyte (CTL) from PBMC cultured on dendritic cells (DC) transfected with hepatocellular carcinoma (HCC) mRNA for adoptive immunotherapy of HCC.Methods DCs were generated from PBMCs.HCCmRNA was amplified from HepG-2 cells and tumor tissuses of 3 cases of HCC by using PCR.Expansion culture of CTL was achieved using PBMCs cultured on DCs transfected with HCCmRNA.Cytotoxicity was measured using a standard 51Cr release assay.The proportion of CD3 +,CD4 +,CD8 + cells was determined using flow cytometry.Results After PBMCs of HCC were cultured on DCs transfected with HepG-2 mRNA or HCCmRNA of 3 HCC petients for 3 weeks,CD3 +,CD8 + cells comprised viz 89.3%, 73.6%, 86.8%, and 53.6% of the CTL,respectively.The CTL from DCs transfected with HepG-2 cells and 2 AFP(+) patient mRNA of killed 91%,90%,and 93% of HepG-2 cells at an E/T ratio of 8 for 24h in vitro.However,the CTL from DCs transfected with HCC(-)patient mRNA killed 46% of HCC HepG-2 cells only at an E/T ratio of 8 for 24 h in vitro.The cytotoxic activity was inhibited by the treatment with anti-CD3,anti-CD8,and anti-MHC-class I monoclonal antibodies but not with anti-CD4 and anti-MHC-class Ⅱ antibodies.Conclusion HCCmRNA-transfected DCs may represent a broadly applicable vaccine strategy to induce potentially therapeutic CTL responses in HCC.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2005年第4期432-434,共3页
Chinese Journal of Experimental Surgery
基金
广东省自然科学基金资助项目 (0 2 1 889)
