四种热休克蛋白／肽混合物预防小鼠肉瘤发生、发展的作用研究  

Anti-tumor activity of the mixture of four heat shock proteins/peptides in mice model for sarcoma

作　　者：彭江[1] 郭全义[1] 袁玫[1] 崔雪梅[1] 汤宇[1] 眭翔[1] 陈继营[1] 许文静[1] 卢世璧[1] 

机构地区：[1]解放军总医院骨科研究所,北京100853

出　　处：《中华骨科杂志》2009年第10期974-979,共6页Chinese Journal of Orthopaedics

基　　金：国家高科技863计划（2007AA021806）

摘　　要：目的探讨四种热休克蛋白（heat shock protein，HSP）／肽混合物防治小鼠肉瘤的作用及其机制。方法小鼠S180肉瘤及MCA．207肉瘤组织，分别剪碎，裂解，离心，取上清上Sephacryl S-200HR层析柱，截取相对分子质量相当于50×10^3-200×10^3段，再经Con A、ADP亲和层析，SDS—PAGE电泳及Western—blot鉴定。分别用HSP-70、HSP-60及热休克蛋白／肽混合物（HSP-70、HSP-60、HSP-110、Gp-96）免疫小鼠，观察无瘤生存率、抑瘤生长率及存活期。用流氏细胞仪检测“T”细胞亚群，ELISPOT检测IFN-1，乳酸脱氢酶检测混合淋巴细胞杀伤作用等方法观察免疫功能的变化。结果HSP-70、HSP-60及热休克蛋白／肽混合物均有预防性免疫治疗效果，热休克蛋白／肽混合物防治S180肉瘤达到41.2％的无瘤长期生存，抑瘤生长率为83．3％，优于单一的HSP亚型。热休克蛋白／肽混合物防治MCA肉瘤达到50．0％的无瘤长期生存，抑瘤生长率为79．0％；多种方法检测表明热休克蛋白／肽混合物疫苗可诱发小鼠的细胞免疫功能。结论热休克蛋白／肽混合物可诱发机体细胞的免疫功能，其抑制肉瘤生长效果优于单一的HSP-70或HSP-60，具有临床应用前景。Objective To investigate the anti-tumor effects of the mixture of four heat shock protein/ peptides in mouse model for sarcoma. Methods The mixture of four subtype heat shock protein/peptides were extracted from mouse sarcoma cell line S180 and MCA-207 by molecular chromatography Sephaeryl S- 200HR and identified by SDA-PAGE and Western blot. Balb/c mice were immunized by the mixed heat shock protein/peptides with different dose, and then S180 and MCA-207 sarcoma cell line was grafted after vaccine in Balb/c or C57 mice. The disease-free survival and tumor growth inhibition rates were examined. Immune response eliciat by heat shock protein/peptides were detected. Sub-group of T lymphocyte was assessed by FACS; CTL by lactate dehydrogenase release assay and IFN-γ by ELISPOT assay. Results The mixture of heat shock protein/peptides and HSP-60, HSP-70, Gp-96 were isolated by chromatography and affinity chromatography separately. The mice vaccinated by the mixture of heat shock protein/peptides has the most protective results. In S180 model, the disease-free survival for tumor was 41.1%, the tumor growth inhibition rates was 83.3%. In MCA-207 model, the disease-free survival was 50.0%, the tumor growth inhibition rates was 79.0%. After immunization, CD8^＋, Nk cell, and IFN-γ were all increased. CTL was 42.4% （B/T=40）. Conclusion These animal vaccination/immunization strategies show that the mixture of heat shock protein/peptides as vaccine can induce immuno-activity against sarcoma and reduce tumor burden prophylactically, it may be possible to develop a specialized sarcoma vaccine for clinical treatments.

关 键 词：肉瘤 免疫疗法 疫苗 热休克蛋白质类 

分 类 号：R686[医药卫生—骨科学]