高表达HSP70鼠脑胶质瘤细胞瘤苗的体外抗瘤作用  

The anti-tumor effect of HSP70 enriched C6tumor cell vaccine in vitro

作　　者：康德智[1] 林元相[1] 苏东辉[2] 何理盛[1] 连葆强[1] 

机构地区：[1]福建医科大学附属第一医院神经外科,福建福州350005 [2]福建医科大学免疫学教研室,福建福州350004

出　　处：《中华神经医学杂志》2003年第1期52-56,共5页Chinese Journal of Neuromedicine

基　　金：福建省教育厅科研基金(01A044)

摘　　要：目的观察高表达热休克蛋白70(HSP70)鼠脑胶质瘤细胞(C6细胞)瘤苗在体外的主动免疫抗瘤效应,并探讨其可能的抗瘤机理。方法用灭活经诱导高表达HSP70的C6细胞作瘤苗,体外刺激SD大鼠脾细胞,进行肿瘤杀伤试验。用MTT法检测其杀伤活性,FCM及电子显微镜观察杀伤瘤细胞的变化。结果1.经C6细胞瘤苗刺激的大鼠脾细胞较直接用灭活C6细胞刺激的大鼠脾细胞或新鲜大鼠脾细胞对C6细胞的杀伤率显著增高,其中经C6瘤苗刺激6 d后的大鼠脾细胞的杀伤率最高,它对热休克型和野生型C6细胞的杀伤率分别达99.25 %和63.10 %(E ∶T=20∶1)。这种杀伤活性可被鼠HSP70单抗所阻断。2.杀伤试验时,FCM检测到亚二倍体峰,电镜发现靶细胞出现凋亡小体。结论高表达HSP70的C6细胞瘤苗,在体外有显著的免疫原性,可激活大鼠脾细胞对同源C6细胞产生有效的杀伤。通过使靶细胞凋亡可能是这种瘤苗主动免疫抗瘤效应的一个重要的杀伤机制。Objective To study the anti-tumor effect and mechanisms of the SD rat splenic cells vaccinated with HSP70 enriched C6 tumor cell vaccine (TCV) in vitro. Methods Mitomycin C-inactivated C6 cells with enriched membrane HSP70 obtained under optimum heat shock conditions were used as stimulants for their co-incubation shock treated C6 cells or their wild-type originals. The cytotoxicity was evaluated by MTT assay. Apoptosis of the target cells was analyzed by flow cytometry and electronic microscopy. Results The specific cytotoxicity of effective cells elicited by HSP70-rich C6 cells vaccine was shown to be significantly higher than that of fresh splenic cells or that of the splenic cells elicited by inactivated wild-type C6 cells in vitro. On the other hand, heat-treated C6 cells, as compared with their wild-type originals, were more sensitive to the effective cell killing. The peak level of cytotoxicity of splenocytes stimulated by these TCV for 6 days against the same heat-treated C6 cells reached 99.25 %, against the wild type cells, 63.10% (E ∶T=20∶1). The cytotoxicity was blocked with anti-HSP70 McAb. Further analysis using FCM and electronic microscopy revealed that apoptosis of target cells was the key form of the cell killing resulting from the HSP70-rich TCV stimulation. Conclusion HSP70-rich C6 TCV can improve its immunogenicity. The anti-tumor effect of the splenic cells stimulated with HSP70-rich C6 TCV can be greatly enhanced, possibly by the apoptosis-mediated mechanism.

关 键 词：脑肿瘤 疫苗 C6细胞 HSP70 主动特异性免疫 

分 类 号：R739.41[医药卫生—肿瘤]