摘要
目的观察高免疫原性人多形性胶质母细胞瘤(GBM)U251细胞疫苗体外诱发产生效应细胞杀伤靶细胞的超微结构变化。方法以500U/mL干扰素-γ(IFN-γ)诱导48h+热43℃休克2h诱导膜型热休克蛋白70(HSP70)及主要组织相容性抗原复合体I类分子(MHC-I)双高表达,经丝裂霉素(MMC)灭活制成高免疫原性细胞疫苗。体外刺激健康捐献者外周血单个核细胞(PBMCs)作为效应细胞(MHC-HSP-CTL),进行肿瘤特异性杀伤试验;MTT比色法检测其杀伤活性;透射电镜观察靶细胞受攻击后情况。结果MHC-HSP-CTL对野生型U251细胞特异性的杀瘤活性明显高于HSP70分子单高表达的细胞疫苗刺激组(HSP-CTL)、MHC-I类分子单高表达的细胞疫苗刺激组(MHC-CTL)以及野生型对照组(W-CTL)(P<0.05),透射电镜显示靶细胞受到攻击后出现不同程度凋亡样坏死和胀亡样坏死,胀亡样坏死细胞在高免疫原性细胞疫苗刺激的效应细胞组较多。结论介导细胞胀亡是高免疫原性即膜型HSP70和MHC-I类分子双高表达疫苗的U251细胞疫苗一种重要主动特异性抗瘤机制。
Objective To examine the uhrastructural changes of target ceils killed by effector cells stimulated by GBM U251 cell vaccine with high immunogenicity in vitro. Methods U251 cells were treated with 500U/mL IFN-γ for 48 hours and heat shock at 43℃ for 2 hours,then inactivated by mitomycin C. PBMCs from healthy donator were incubated with GBM U251 cell vaccines with high immunogenicity, then collected as effector cell named MHC-HSP-CTL. The competence of the effector cell was determined by MTT assay; the uhrastructure of the attacked target cells was observed by Electron microscope. Results MHC-HSP CTL were more competent in specifically killing wild type U251 cells than those stimulated either by U251 cell vaccine with membrane-enriched HSP 70（HSP-CTL）or by U251 cell vaccine with membrane-enriched MHC class I （MHC-CTL）, or by untreated control group（W-CTL）（P〈0.05）; A distinct extent of both apoptotic necrosis and oncotic necrosis can be observed in the target cells, with more cells undergoing oncotic death when treated with effector cells stimulated by U251 cell vaccine with high immunogenicity than the other two groups. Conclusion The oncotic necrosis of the target cells induced by the effector cells which were stimulated by the GBM U251 cell vaccine with membrane-enriched HSP 70 and MHC class I molecule may be an important mechanism underlying its antitumor activity
出处
《福建医科大学学报》
2009年第2期110-113,共4页
Journal of Fujian Medical University
基金
福建省科技计划项目(K04059)
福建医科大学科学研究发展基金(FJGXY04028)
