跨膜型超抗原SEA融合蛋白的表达及其对小鼠黑色素瘤的免疫治疗作用  

Expression of the fusion protein of transmembrane-superantigen SEA and its antitumor effects

作　　者：马文学[1] 余海[2] 王青青[2] 鲍建芳[3] 黄常新[2] 李经忠[2] 金洪传[1] 陈海祥[3] 邱翔[3] 

机构地区：[1]浙江大学肿瘤研究所,杭州310009 [2]浙江大学免疫学研究所,杭州310006 [3]浙江大学医学院细胞与分子生物学实验中心

出　　处：《中华微生物学和免疫学杂志》2003年第7期567-571,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目 ( 3 9770 83 7);浙江省自然科学基金资助项目 ( 3 9913 1;3 0 15 80 )

摘　　要：目的　制备跨膜型超抗原融合蛋白 ,研究该蛋白制备的肿瘤疫苗的抗肿瘤作用。方法用已构建的重组跨膜型超抗原的表达载体pET 2 8a TM SEA转化E .coliBL2 1(DE3)pLysS宿主菌 ,在IPTG诱导下产生目的蛋白 ;用Ni NTAHisBindResin纯化带有His Tag的目的蛋白。采用免疫组化法、免疫荧光法和流式细胞术检测其对细胞膜的锚定作用。建立C5 7BL 6小鼠的黑色素瘤模型 ,观察跨膜型超抗原SEA融合蛋白制备的肿瘤疫苗对荷瘤小鼠的免疫治疗作用。结果　跨膜型超抗原融合蛋白能够锚定在肿瘤细胞膜上。融合蛋白制备的肿瘤疫苗能够显著抑制荷瘤小鼠肿瘤的生长 ,并延长其生存期。结论　跨膜型超抗原融合蛋白制备的肿瘤疫苗具有较强的抗肿瘤作用 ,有可能作为一种有效的新型疫苗用于肿瘤的治疗和预防。Objective To prepare a purified fusion protein of transmembrane superantigen SEA and explore its antitumor effects. Methods The constructed recombinant expression vector pET-28a-TM-SEA was transformed into E.coli BL21(DE3)pLysS strain. The target protein was induced at 30℃ with 1mM of IPTG. The target protein with His-Tag was purified by Ni-NTA His Bind Resin purification system. The anchoring effect was identified by immunohistochemistry, immunofluorescence method and flow cytometric assay after co-incubation of this fusion protein with Colo-205 cells or B16 cells for 4 hrs. The melanoma model of the C57BL/6 mice was established, the antitumor effect of immunotherapy with the tumor vaccine prepared with TM-SEA fusion protein were investigated. Results The purified TM-SEA fusion protein anchored into the membrane of tumor cells. After being treated with the tumor vaccine based on TM-SEA fusion protein, tumor growth was significantly inhibited and the survival period of the tumor-bearing mice was longer as compared with the control groups. Conclusion The tumor vaccine made by TM-SEA fusion protein significantly inhibited the tumor growth in melanoma-bearing mice. This result is clinically potential for the treatment of tumor recurrence and micro-metastasis.

关 键 词：肿瘤疫苗 跨膜型超抗原融合蛋白 黑色素瘤 免疫治疗 金黄色葡萄球菌肠毒素 

分 类 号：R392[医药卫生—免疫学]