Tca8113-4-1BBL细胞联合CD28单抗在口腔鳞癌免疫治疗中的作用

Antitumor immunotherapy induced by Tca8113-4-1BBL cell combined with anti-CD28 mAb:An in vitro study

目的:探讨转4-1BBL肿瘤细胞联合抗CD28单抗体外诱导抗肿瘤活性的能力。方法:通过脂质体法将重组载体pEGFP/neo-h4-1BBL转染人舌鳞癌细胞Tca8113,经G418(400μg/mL)筛选及有限稀释后,获得稳定高表达克隆,分别用RT-PCR和Western印迹检测转染细胞中h4-1BBLmRNA和蛋白的表达。将转染与未转染4-1BBL基因的Tca8113细胞用丝裂霉素C(MMC)处理后,制成肿瘤细胞瘤苗。联合抗CD28单克隆抗体与经体外抗CD3mAb诱导的人外周血T淋巴细胞共同培养,测定T细胞增殖、CTL杀伤活性及产生细胞因子(IL-2和IFN-γ)的能力。实验数据以SPSS12.0软件包进行方差分析。结果:h4-1BBL基因真核表达载体在Tca8113细胞中获得稳定表达。转染h4-1BBL基因的Tca8113细胞联合抗CD28单抗能显著刺激T细胞活化、增殖(P<0.01),促进IL-2、IFN-γ分泌(P<0.01),并能有效地诱导CTL的特异性杀伤活性(P<0.01)。结论:转4-1BBL肿瘤细胞联合抗CD28抗体能显著增强肿瘤细胞的免疫原性,诱导T细胞产生有效的抗肿瘤免疫应答。

PURPOSE： To explore the effect on activation and cytotoxicity of human peripheral blood T lymphocyte induced by anti-CD28 mAb with h4-1BBL gene transfected into Tca8113 cells in vitro. METHODS： The eukaryotic expression vector pEGFP/neo-h4-1BBL was transfected into human tongue squamous cell carcinoma cells line Tca8113 by lipofectamine2000. Then the transfected cells were selected in medium containing G418 （400μg/mL）, cloned by limited dilution and named as Tca8113-4-1BBL. Human 4-1BBL mRNA and protein expression of transfected cells was detected by RT-PCR and Western blot, respectively. The tumor cell vaccines （TCV）were obtained by treatment with mitomycin （MMC）. The TCV were co-cultivated with anti-CD3 activated T lymphocytes with or without anti-CD28.Then the cytotoxic activity of CTL and the cytokines production （IFN-γ and IL-2） were tested. SPSS12.0 software package was used for statistical analysis. RESULTS： The Tca8113 cells transfected by pEGFP-h4-1BBL could express human 4-1BBL efficiently. As compared with wild typical Tca8113 cells, the anti-CD28 mAb with transfected Tca8113 cells had more effect for proliferation, IL-2, IFN-7 production and cytotoxie activity of lymphocytes. CONCLUSIONS： The anti-CD28 mAb with Tca8113 ceils transfected with 4-1BBL is effective in enhancing its immunogenicity and inducing antitumor immune response.