人脑胶质瘤特异性细胞毒淋巴细胞的诱导及抗瘤活性的实验研究  

Preperation of tumor specific cytotoxic T lymphocytes in glioma and observation of its anti-tumor effect

作　　者：马文雄[1] 陈桂林[1] 姜文学[1] 吴士良[2] 谢学顺[1] 徐庚达[1] 吴杰[1] 韦永新[1] 惠国桢[1] 

机构地区：[1]苏州大学附属第一医院,215006 [2]苏州大学附属第一医院生化教研室,215006

出　　处：《江苏医药》2001年第11期820-822,826,I002,共5页Jiangsu Medical Journal

摘　　要：目的　制备人脑胶质瘤特异性细胞毒T淋巴细胞 ,并对其增殖能力 ,抗瘤活性及作用机制方面等进行实验研究。方法　 (1)运用蛋白质分离、纯化技术 ,成功地从SHG 44细胞中提取出能刺激胶质瘤患者外周血淋巴细胞 (PBMC)的有效成份 ,利用该抗原与低剂量IL 2、CD3 单抗共同刺激胶质瘤患者PBMC ;并采用3 H TdR掺入试验 ,对HGS CTL、TIL和CD3 AK细胞的增殖活性和细胞毒性进行动态观察。 (2 )通过ELISA方法检测上述三种效应细胞分泌细胞因子水平。 (3)对NHG 1荷瘤鼠的治疗观察。结果　 (1)HGS CTL的增殖活性始终明显高于TIL和CD3 AK细胞 ,且具有增殖快 ,维持时间长等特点。HGS CTL特异性杀伤活性明显高于TIL和CD3 AK细胞 (P <0 0 5 )。FACS测定表型为以CD+ 3 、CD+ 8CTL为主的异质细胞群。 (2 )HGS CTL分泌TNF α ,IL 2水平明显高TIL和CD3 AK细胞 (P <0 0 5 ) ,IL 8的分泌水平无明显差异 (P >0 0 5 )。 (3)NHG 1荷瘤鼠的治疗观察 ,局部用药组瘤重 0 32± 0 11g ,静脉用药组瘤重 0 5 8± 0 18g ,与其对照组瘤重 0 97± 0 45 g和 0 98± 0 17g相比 ,差异显著 (P <0 0 5 )。 结论　以上实验结果表明 ,HGS CTL在体内外均具有显著抗瘤活性 ,具有增殖快、制备简便等特点 ,因此将成为胶质瘤患者较有效的辅助?Objective This article describes the induction of human glioma specific cytotoxic T lymphocytes(HGS-CTL) in vitro.Experiments were done for the evaluation of its proliferetive ability and the mechanism of anti-tumor effect both in vitro and in vivo.Methods (1)An effective component was successfully obtained from human glioma cell line SHG-44 by protein separation and purification techniques,which can activate the peripheral blood lymphocytes(PBMC) of the patients with glioma.Combined infusion of glioma-associated antigens,low dose of IL-2 and anti-CD 3 MCAb can activate the PBMC of the patients with glioma.The propagation capacity and cytotoxic effects of the HGS-CTL,TIL and CD 3AK cells were continuously observed by means of the standard 3H-TdR technique.(2)The levels of cytokines secreted by the above three effector cells were measured by ELISA.(3)The curative effect of HGS-CTL was observed on NHG-1.Results (1)The propagation capacity of HGS-CTL was obviously higher than any others and characterized by rapid progagation and long duration.So was the specific killing activity of HGS-CTL(P<0.05).The HGS-CTL thus obtained was mainly composed of heterogeneous cell groups with CD + 3,CD + 8CTL predominated by FACS.(2)The TNF-a and IL-2 secreted by HGS-CTL were significantly higher than that of TIL and CD 3AK cells(P<0.05).There was no obvious difference in IL-8(P>0.05).(3)From the treatment of NHG-1 tumor leading mice,it was found that the group of local use(0.32±0.11g) and the group of intravenous use(0.58±0.18g) demonstrated marked changes(P<0.05),as compared with the tumor weights(0.97±0.45g)and (0.98±0.17g)in the controls.Conclusion The results mentioned above show that HGS-CTL has high specific anti-tumor activity,either in vitro or in vivo.It has characteristi

关 键 词：脑胶质瘤 细胞毒T淋巴细胞 肿瘤抗原 免疫治疗 实验研究 

分 类 号：R739.41[医药卫生—肿瘤]