转录因子T-bet表达增强人树突状细胞疫苗诱导的抗肝癌免疫

Transcription factor T-bet enhances dendritic cell-induced immunity against hepatocellular carcinoma

目的:探讨T-bet在肝癌患者外周血来源的树突状细胞(dendritic cells,DCs)中表达能否增强其诱导抗肿瘤免疫。方法:取肝癌患者外周血单核细胞,用5μg/L rhGM-CSF、5μg/L rhIL-4培养6d成不成熟DC(iDC),随后加10μg/L TNF-α诱导成熟DC。用冻融法制备肝癌细胞株HepG2肿瘤抗原,致敏DC,并分组如下:loaded DC/TNF-α(loaded mDC);loaded DC/TNF-α+IFN-γ(loaded DC/T+I);loaded DC/T-bet(loadedDC/T-bet);iDC。体外刺激淋巴细胞。观察T-bet外源表达对DC的表型、混合淋巴细胞反应、肿瘤特异性细胞杀伤效率影响。结果:外源表达T-bet促进DC/T-bet表型成熟,促进自体混合淋巴细胞反应,诱导分泌出更多的Th1型细胞因子,增强肝癌细胞特异性杀伤效应。结论:T-bet增强DC抗肿瘤免疫性能。

AIM ： To observe that the ectopic expression of transcription factor T - bet in dendritic cells （DC） enhances anti - tumor immunity. METHODS. Peripheral blood mononuclear ceils were isolated from patients with primary hepatocellular carcinoma （HCC）. The monocytes were stimulated with GM - CSF, IL - 4 and TNF - α for inducing the mature DC. Tumor associated antigens （TAA） of HepG2 was generated with freeze - thawing methods and pulsed DC. Groups was arranged as followed： loaded DC/TNF - α（ loaded mDC）, loaded DC/TNF -α ＋ INF -γ （ loaded DC/T ＋ I）, loaded DC/T - bet （ loaded DC/T - bet）, iDC. Lymphocytes were stimulated with DC described as above. The phenotype of DC, MLR （mixed lymphocyte reaction）, and tumor- specific cytotoxicity of CTL （cytotoxic T lymphocytes） were observed. RESULTS： Compared with control, elevated expression of HLA - DR, CD80, and CD86, enhanced second MLR （mixed lymphocyte reaction）, up -secretion of Thl -type cytokines, and more potent ability to induce CTL specific toxicity to human hepatic carcinoma HepG2 were observed in DCs with ectopic expression of T - bet. CONCLUSION： DCs with ectopic expression of T - bet enhance anti - tumor immunity.