中文摘要：

目的构建转染人口腔鳞状细胞癌细胞株4-1-BBL基因，探讨其体外诱导抗肿瘤免疫的能力。方法通过脂质体法将重组载体pEGFP-4-1-BBL转染人口腔鳞状细胞癌细胞Tca8113，经G-418筛选及有限稀释后获得稳定高表达克隆，分别用反转录聚合酶链反应（RT-PCR）和蛋白质印迹法检测转染细胞中人4-1-BBL mRNA和蛋白的表达。制备肿瘤细胞疫苗。用淋巴细胞分离液分离纯化人外周血T淋巴细胞，用抗CD-3单抗预处理T细胞后，与转染及未转染人4-1-BBL的Tca8113疫苗混合培养。用CCK-8比色法检测细胞毒性T淋巴细胞（cytotoxic T lymphocytes，CTL）的杀伤活性；锥虫蓝计数法检测T细胞增殖，酶联免疫吸附实验检测培养上清液中白细胞介素（IL）-2、干扰素γ分泌水平。结果转基因Tca8113细胞能够稳定高表达人4-1-BBL。与野生型的Tca8113细胞相比较，转染人4-1-BBL的Tca8113细胞PCR扩增产物的大小约271bp，能够显著增强T细胞增殖，促进IL-2、干扰素γ分泌，并能有效地诱导CTL产生对Tca8113细胞的特异性杀伤作用。结论转染4-1-BBL基因既能增强野生型Tca8113细胞的免疫原性，也能诱导T细胞产生有效的抗肿瘤免疫应答。

英文摘要：

Objective To examine the activation and cytotoxicity of human peripheral blood T lymphocyte induced in vitro by human 4-1-BB ligand（4-1-BBL） gene transfected into tumor Tca8113 cells. Methods The eukaryotic expression vector pEGFP-h4-1-BBL was transfeeted into human oral carcinoma cell line Tea8113 by LipofectamineTM2000. The transfeeted cells were then selected in medium containing G- 418, cloned by limited dilution and named as Tca8113-4-1-BBL. Human 4-1-BBL mRNA and protein expression of transfected cells was detected by reverse transcriptase-polymerase chain reaction （RT-PCR） and western blotting respectively. The tumor cell vaccines （TCV） were obtained by treatment with mitomycin （MMC）. Human peripheral blood mononuclear cells （PllMC） were prepared from lymphoprep, and then stimulated with anti-CD-3 mAb and incubated with non-transfeeted or transfected TCV-Tca8113 cells, respectively. The proliferation of T cells was evaluated by trypan blue exclusion; the CCK-8 was used to detect the eytotoxie effect of T lymphocytes. Meanwhile, the secretion of interferom-~/ （IFN-γ） and interleukin（IL）-2 in culture supernatant was detected by enzyme-linked immunosorbnent assay （ELISA）. Results The Tca8113 cells transfected by pEGFP-h4-1-BBL could express human 4-1-BBL efficiently. As compared with wild type Tca8113 cells, the transfccted TcaS113 cells could markedly promote proliferation, IL-2 and IFN-γ production and cytotoxic activity of lymphocytes. Conclusions The transfeetion of human 4-1-BBL gene in Tca8113 cells is effetive in enhancing its immunogenicity and inducing antitumor immune response in vitro.