摘要
目的观察联合激动Toll样受体(TLR)能否使树突细胞(DC)达到更理想的成熟状态,并诱导出抗结肠癌免疫反应。方法在体外培养DC过程中加入TLR3和TLR7/8配体,流式细胞仪检测DC重要表型表达,酶联免疫吸附实验(ELISA)法检测白细胞介素(IL)-12分泌量,3H—thymidine掺人法检测DC促淋巴细胞增殖能力,使用CEA转基因鼠和转染CEA的MC38小鼠结肠癌细胞株(MC38-CEA)构建动物模型,乳酸脱氢酶(LDH)释放法检测DC诱导细胞毒性T淋巴细胞(CTL)对MC38-CEA的体外杀伤作用。结果联合激动TLR的DC与常规组比较,能高水平表达DC重要表型,分泌更多IL-12(P〈0.01),更有效刺激淋巴细胞增殖(P〈0.01),荷载抗原后诱导的效应细胞在效靶比为20:1和40:1时,对MC38-CEA的杀伤率为(31.9±3.3)%和(39.1±4.2)%,而其他各组无明显杀伤作用(P〈0.05)。结论联合激动TLR的DC具有更理想的成熟度,能诱导更有效的抗肠癌免疫反应,提示联合激动TLR可能是增强DC肿瘤疫苗疗效的有效途径。
Objective To investigate the immune functions of DCs with synergistic activation of TLR ligands and the antitumor effects against colon cancer induced by DCs in vitro. Methods We involved combination of TLR ligands in culture protocol of DCs. The DCs' important surface molecules expression was detected by flow cytometry. DCs'IL-12 secretion was evaluated by ELISA and the proliferating capacity of lymphocytes was tested by 3 H-thymidine assay. The CEA transgenic mice and MC38-CEA mouse colon cancer cell line were employed as animal model and the CTL activity against MC38-CEA induced by pulsed DCs was tested by LDH cytotoxicity assay. Results Compared with conventional generated DCs, DCs combined with activation of TLR showed higher surface molecules expression ,significantly higher secretion of IL-12 and more potent proliferating capacity of lymphocytes as well. The cytolytic activity against MC38-CEA cells shown by effector cells induced by pulsed DCs with combined activation of TLR was (31.9 ±3.3 )% and (39.1 ±4. 2)% when the E: T ratio was 20:1 and 40:1 respectively, but no obvious cytolytic activity was found showed in other groups. Conclusion DCs combined with activation of TLR showed better maturation status and can also induce more effective antitumor immune effects against colon cancer, suggesting combined activation of TLR may be the effective strategy for developing more powerful DCs cancer vaccines.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2010年第3期353-355,共3页
Chinese Journal of Experimental Surgery
关键词
结肠肿瘤
树突细胞
TOLL样受体
癌胚抗原
Colonic neoplasms
Dendritic cells
Toll-Like receptors
Carcinoembryonic antigen