负载gp96－Ki67 280-288复合物的树突状细胞疫苗对骨肉瘤的抗肿瘤效应体内实验研究  

Effects of dendritic cell vaccine loaded with gp96-Ki67280-288 complexes on mice with osteosarcoma

作　　者：严专[1] 李慧忠 许铁[3] 刘林[3] 

机构地区：[1]徐州医学院研究生学院2012级,江苏 徐州221004 [2]江苏省肿瘤生物治疗研究所,江苏 徐州221002 [3]徐州医学院急救与救援系,江苏 徐州221004

出　　处：《徐州医学院学报》2015年第6期402-406,共5页Acta Academiae Medicinae Xuzhou

基　　金：国家青年科学基金（81202015）;江苏省高校自然科学基金面上项目（12KJB320014）;徐州市科技课题（XM128025）

摘　　要：目的：研究HSPgp96与Ki67280－288复合物在体内的特异性抗肿瘤作用机制。方法设计分组对照实验，分别给予gp96－Ki67280－288复合物、Ki67280－288多肽、HSPgp96、PBS免疫HLA－A倡0201转基因小鼠。 MTT法检测小鼠淋巴细胞增殖情况，ELISA法检测其诱导的细胞毒性T细胞（ CTL）分泌干扰素（ IFN）－γ水平，乳酸脱氢酶（LDH）释放试验检测所诱导的CTL对U2OS骨肉瘤细胞的杀伤活性，同时在U2OS骨肉瘤动物模型上观察gp96－Ki67280－288复合物修饰后的树突状细胞（DC）所诱导产生的CTL对肿瘤生长及动物生存期的影响。结果gp96－Ki67280－288复合物免疫的小鼠的淋巴细胞表现出最强的增殖能力，淋巴细胞上清液中的IFN－γ水平较其他组升高（P＜0．05）；LDH释放实验表明，随着效靶比的提高，各组淋巴细胞对U2OS细胞的杀伤率相应增高（P＜0．05），在效靶比为40∶1的时候最高，gp96－Ki67280－288组的杀伤率高于HSPgp96组、Ki67组和PBS组（P＜0．05）。荷瘤裸鼠实验结果显示，从第12天以后，gp96－Ki67280－288组小鼠肿瘤体积明显小于其他组（P＜0．05）；gp96－Ki67280－288组的抑瘤率高于HSPgp96组和Ki67组（P＜0．05），较其他3组表现出更长的生存期（P＜0．05）。结论 gp96－Ki67280－288复合物能很好地修饰树突状细胞，使树突状细胞有效地提呈抗原，并诱导抗原特异性抗肿瘤免疫反应，具有很强的体内抗肿瘤作用。Objective To investigate the anti-tumor activity of the complex of HSPgp96 and HLA-A2-restricted epitope Ki67280-288 in vivo.Methods HLA-A＊0201 transgenic mice were divided into four groups according to their treatment of gp96-Ki67280-288 complex, Ki67280-288 peptide, HSPgp96 or PBS.Then, the proliferation of lymphocytes in the mice was determined by MTT.The level of IFN-γsecreted by cytotoxic T lymphocytes （CTLs） induced by the com-plex was measured by ELISA.The cytotoxicity of CTL on U2OS cells were detected by LDH release assay.Meanwhile, the effects of CTL induced by DCs loaded with gp96-Ki67280-288 complex on tumor growth and survival were examined u-sing nude mice bearing U2OS osteosarcoma.Results Lymphocytes from the mice loaded with gp96-Ki67280-288 com-plex grew at the fastest rate growth, and produced the highest levels of serum IFN-γ（P〈0.05）.According to LDH re-lease assay, the cytotoxicity of CTL on U2OS cells were enhanced as the effector-to-target ratio increased （P〈0.05）, and reached the top at an effector-to-target ratio of 40∶1, when stronger cytotoxicities were measured in Group gp96-Ki67280-288 complex than the other three group （P〈0.05）.Meanwhile, injection of gp96 -Ki67280-288 complex in to nude mice bearing U2OS osteosarcoma could remarkably inhibit the growth of the tumor and longer survival when com-pared with the other three treatments from Day 12 （P〈0.05）.Conclusion The gp96-Ki67280-288 complex can modify DCs and make them present antigens to T cells in an effective manner.It can also induce antigen-specific anti-tumor immunity, producing potent activities against tumor in vivo.

关 键 词：树突状细胞 热休克蛋白 KI67 骨肉瘤 抗肿瘤免疫 

分 类 号：R738.7[医药卫生—肿瘤]