rhHSP70联合冻融抗原修饰树突状细胞诱导的抗乳腺癌作用  

Specific anti-tumor immune responses of dendritic cells pulsed with recombinant human rhHSP70 and freeze-thaw cellular lysates derived from breast cancer

作　　者：李斌[1] 陈鹏[2] 郑建云[3] 

机构地区：[1]西安医学院附属医院肿瘤外科,西安市710077 [2]西安医学院生化教研室 [3]西安医学院附属医院病理科

出　　处：《中国肿瘤临床》2013年第15期888-892,共5页Chinese Journal of Clinical Oncology

基　　金：陕西省教育厅课题(编号:2010JK806);西安医学院校级课题(编号:2007FZ11)资助~~

摘　　要：利用rhHSP70联合树突状细胞递呈肿瘤抗原的特性提高细胞毒T淋巴细胞(CTLs)对乳腺癌细胞的杀伤活性。方法:外周血单个核细胞体外经GM-CSF和IL-4诱导产生树突状细胞,负载冻融抗原肽的同时加入新型热休克蛋白(rhHSP70),不同分组分别诱导自体CTLs产生。ELISA测定CTLs杀伤活性和细胞因子的分泌。结果:冻融抗原肽致敏的DCs促进CTLs增殖,上调CTLs中CD3+和CD8+T细胞群及Th1型细胞因子的分泌;体外实验中具有对人乳腺癌细胞MCF-7的杀伤活性,在加入rhHSP70后效果更加明显,并能显著增强CTLs对肿瘤细胞的杀伤率。结论:rhHSP70联合肝癌冻融抗原修饰DCs,能够促进DCs的成熟,增强DCs刺激淋巴细胞增殖的能力,诱导的CTLs在体外对乳腺癌细胞能产生高效杀伤力。rhHSP70增强DCs抗肿瘤能力的机制可能与其促进DCs成熟有关。Objective： This work aims to use the characteristics of dendritic cells （DCs） pulsed with recombinant human HSP70, which can present and process tumor antigens, to enhance the killing activity of cytotoxic t lymphocytes （CTLs） against breast neoplasms. Methods： Autologous DCs were isolated from peripheral blood mononuclear cells and then stimulated in vitro with granulocyte macrophage-colony stimulating factor and interleukin-4. The DCs were loaded with A549 tumor cell freeze-thaw lysate, and rhHSP70 was added as an immune adjuvant. The specific groups were subjected to tumor-specific cytotoxic assay, enzyme-linked immunosorbent assay, and fluorescence-activated cell sorting. Results： DCs pulsed with A549 tumor cell lysate enhanced the growth expansion of CTLs, upregulated CD40 and CD80 populations in CTLs, and augmented Thl cytokines. In addition, the cytotoxicity of specific CTLs against A549 was highly enhanced. The above indications became more obvious after the addition of rhHSP70. Conclusion： DCs pulsed with freeze-thaw cell lysates derived from breast cancer can enhance growth expansion of lymphocytes. They may serve as an effective tumor antigen to stimulate the proliferation of specific CTLs, which are very effective in activating specific T-cell responses against breast cancer cells in vitro. The improved anti-tumor immunity response by DC-based vaccines may be related to the maturation of the DCs promoted by rhHSP70.

关 键 词：乳腺癌 T淋巴细胞 热休克蛋白 树突状细胞 肿瘤裂解物 

分 类 号：R392[医药卫生—免疫学]