自身肺癌抗原致敏DCs抗肺癌体外免疫效应研究.docVIP

自身肺癌抗原致敏DCs抗肺癌体外免疫效应研究 　　【摘要】 目的 探讨自身肺癌抗原致敏树突状细胞(Dendritic Cell,DC)的体外免疫效应。方法 体外原代培养肺癌细胞,并制备冻融抗原,冲击DCs,致敏肿瘤浸润淋巴细胞(TIL)和外周血T淋巴细胞,MTT法检测DCs瘤苗体外联合抗肺腺癌细胞效应。结果 采用效靶比为30:1和20:1的比例,30:1和20:1的CTL杀伤率为(18.21±2.89)和(24.69±3.51); 30:1和20:1的CTIL杀伤率为(25.46±3.02)和(27.52±2.68),CTIL比CTL杀伤率高(P0.05)。结论 负载自身肺癌抗原的DCs致敏的肿瘤浸润淋巴细胞TIL比外周血T淋巴细胞杀伤率高,为肺癌的生物治疗提供了新的思路。 　　【关键词】肺癌;树突状细胞;T淋巴细胞;杀伤率 　　 　　Astudy on the specific immunological effect induced by DCs loaded with autospecific lung cancer antigen 　　SHEN Xiao li,ZHAO Jun,WANG Zhi hua,et al.Changzhi 2nd people’s hospital,Shanxi,Changzhi 046000,China 　　【Abstract】 Objective To discuss the specific immunological effect induced by DCs loaded with autospecific lung cancer antigen.Methods Primary lung cancer cells were cultured in vitro and prepared to antigen through thaw and freeze.Antigenwas presented totumor infiltrating lymphocyte(TIL)and T lymphocyte derived from peripheral blood by dendritic cells(DCs).The specific anti tumor immunological effect of DCs vaccine was detected by MTT method.Results 30:1 and 20:1 ratio were employed as effector cell:target cell.The kill rates of CTL with 30:1 and 20:1 were 18.21±2.89 and 24.69±3.51,respectively;the kill rates of CTIL were 25.46±3.02 and 27.52±2.68.The kill rate of CTIL was higher than the CTL(P0.05).Conclusion The kill rate ofCTIL sensitized by DCs loaded autospecific lung cancer antigen is higher than CTL.It provides a new clue for biotherapy of lung cancer. 　　【Key words】 Lung cancer; Dendritic cell; T lymphocyte; Kill rate 　　 　　肺癌为当前世界各地最常见的恶性肿瘤之一,到2005年我国每年肺癌已超过100万,成为世界第一肺癌大国。传统的手术治疗、放疗、化疗虽在不断地改进,但肺癌患者的总体生存率并无明显改善,5年的生存率不足15%[1]。因此,人们在不断探索新的肺癌治疗手段。以树突状细胞(dendritic cell,DC)为基础的肿瘤治疗已成为当今肿瘤生物治疗的焦点之一。本研究以树突状细胞提呈肺癌患者自身肿瘤抗原致敏肿瘤浸润淋巴细胞(tumor infiltrating lymphocyte,TIL)和外周血T淋巴细胞,以评价其抗肺癌效应。 　　1 材料和方法 　　1.1 材料 主要试剂:rhGM CSF,rhIl 4,rhIL 2,Ficoll Hypaque淋巴细胞分离液(中国医学科学院血液学研究所),RPMI 1640(GIBCO),胰蛋白胨(Polypepton),胎牛血清、胰酶(Coring),MTT、胶原酶Ⅳ、dispase酶、DNA酶(Sigma),肺腺癌A549细胞株由郑州大学病理生理教研室惠赠。 　　肿瘤标本的获取:11例肺癌患者均为山西省