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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 To prepare human glioma-specific cytotoxic T lymphocytes and to study its proliferation, anti-tumor activity and mechanism of action. Methods (1) The active components of peripheral blood lymphocytes (PBMCs) that can stimulate glioma patients were successfully extracted from SHG 44 cells by protein separation and purification technology. The antigen was used in combination with low dose of IL 2 and CD3 monoclonal antibody PBMCs were stimulated in glioma patients and the proliferative activity and cytotoxicity of HGS CTL, TIL and CD3 AK cells were observed by 3 H TdR incorporation assay. (2) The levels of cytokines secreted by these three effector cells were measured by ELISA. (3) The treatment of NHG 1 tumor-bearing mice. Results (1) The proliferative activity of HGS CTL was always higher than that of TIL and CD3 AK cells, and had the characteristics of rapid proliferation and long maintenance time. HGS CTL specific cytotoxicity was significantly higher than TIL and CD3 AK cells (P <0 05). The phenotypes determined by FACS were heterogeneous cell populations mainly including CD + 3 and CD + 8CTL. (2) The levels of TNFα and IL-2 secreted by HGS CTL were significantly higher in TIL and CD3-AK cells (P <0.05), while there was no significant difference in IL8 secretion (P> 0.05). (3) The treatment of NHG-1-bearing mice showed that the tumor weight of the local drug-treated group was 0 32 ± 0 11g, the tumor weight of the intravenous drug-treated group was 0 58 ± 0 18g, compared with that of the control group 0 97 ± 0 45 g and 0 98 ± 0 17g compared to the significant difference (P <0 0 5). Conclusion The above experimental results show that HGS CTL has significant anti-tumor activity both in vivo and in vitro, has the characteristics of rapid proliferation, easy preparation and so on, and will therefore be an effective adjunct to glioma patients.