gp96多肽复合物对树突状细胞成熟的影响  

Effects of gp96-peptide complex on maturation of dendritic cells

作　　者：杨威[1] 曹春霞 吴胜利[1] 刘青光[1] 马清涌[1] 潘承恩[1] 

机构地区：[1]西安交通大学第一医院肝胆外科,西安市710061 [2]西安卫生学校

出　　处：《中华肝胆外科杂志》2005年第8期555-558,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然科学基金资助(No.30200369);陕西省科技攻关项目(2002K10-G8)

摘　　要：目的研究热休克蛋白gp96多肽复合物与小鼠骨髓来源的树突状细胞(DC)成熟的关系。方法采用GM-CSF、IL-4刺激培养小鼠骨髓细胞,增殖产生大量DC;从小鼠肝癌细胞株H22细胞中提取gp96肽复合物,体外修饰DC;通过流式细胞仪检测上清液IL-12、TNF-α细胞因子含量和DC表面主要组织相容性Ⅱ类抗原、CD40、CD80等分子表达;DC与小鼠脾淋巴细胞共同培育后,检测CD4、CD8和IFN-γ、IL-10双阳性细胞比例;以51Cr释放法测定细胞毒性T淋巴细胞(CTL)杀伤活性。结果gp96多肽复合物修饰的DC大量分泌IL-12、TNF-α等细胞因子,高表达MHCⅡ类抗原、CD40、CD80等分子;并且能激活小鼠脾淋巴细胞,CD8+-IFN-γ+和CD4+-IFN-γ+双阳性细胞比例显著升高;能够特异性的杀伤H22癌细胞。结论gp96多肽复合物能够诱导DC成熟,体外产生特异性CTL反应。Objective To investigate the relation of gp96-peptide complex to maturation of mouse marrow-derived dendritic cells （DCs）. Methods Mouse bone marrow cells were cultured and stimulated with GM-CSF and IL-4 to obtain plenty of DCs. gp96-peptide complex obtained from H22 liver cancer cells in mice were co-cultured with DCs in vitro. The contents of IL-12 and TNF-α in the supernatant and MHC-II antigen, CD40 and CD80 on DC membrane were determined by flow cytometry. Spleen lymphocytes of mice were activated by modified DCs and the cytotoxicity detected by 51Cr release method. Meanwhile, the alteration of proportions of CD8^＋-IFNγ^＋ and CD8^＋-IL-10^＋ cells, CD4^＋- IFN^γ＋ and CD4^＋-IL-10^＋ cells were analyzed. Results The DCs modified by gp96-peptide complex secreted large amount of IL-12, TNF-α and highly expressed MHC-II antigen, CD40, CD80 on the surface. Spleen lymphocytes of mice were activated and proportions of CD8^＋- IFNγ＋ and CD4^＋- IFNγ^＋ were elevated. The activated and proliferated lymphocytes could specifically kill H22 cells. Conchtsions gp96-peptide complex can induce DC maturation to result in specific CTL response in vitro.

关 键 词：树突状细胞 热休克蛋白gp96多肽复合物 细胞成熟 Balb/c小鼠 热休克蛋白GP96 多肽复合物 特异性CTL反应 小鼠骨髓细胞 流式细胞仪检测 组织相容性Ⅱ类抗原 

分 类 号：R735.7[医药卫生—肿瘤]