CAJ | 학술논문

目的观察联合激动Toll样受体(TLR)能否使树突细胞(OC)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掺入法检测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.
목적 관찰연합격동Toll양수체(TLR)능부사수돌세포(OC)체도경이상적성숙상태,병유도출항결장암면역반응.방법 재체외배양DC과정중가입TLR3화TLR7/8배체,류식세포의검측Dc중요표형표체,매련면역흡부실험(ELISA)법검측백세포개소(IL)-12분비량,~3H-thymidine참입법검측DC촉림파세포증식능력,사용CEA전기인서화전염CEA적MC38소서결장암세포주(MC38-CEA)구건동물모형,유산탈경매(LDH)석방법검측DC유도세포독성T림파세포(CTL)대MC38-CEA적체외살상작용.결과 연합격동TLR적DC여상규조비교,능고수평표체DC중요표형,분비경다IL-12(P＜0.01),경유효자격림파세포증식(P＜0.01),하재항원후유도적효응세포재효파비위20:1화40:1시,대MC38-CEA적살상솔위(31.9±3.3)%화(39.1±4.2)%,이기타각조무명현살상작용(P＜0.05).결론 연합격동TLR적DC구유경이상적성숙도,능유도경유효적항장암면역반응,제시연합격동TLR가능시증강DC종류역묘료효적유효도경.
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 ~3H-thymidine assay.The CEA tramgenic mice and MC38-CEA mmouse 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 aetivation of TLR may be the effective strategy for developing more powerful Dt2s cancer vaccines.