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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 transmembrane-type superantigen fusion protein and study the antitumor effect of the tumor vaccine prepared by this protein. Methods The recombinant E. coli BL21 (DE3) pLysS was transformed into E. coli BL21 (DE3) pLysS by using the constructed transmembrane supervirus expression vector pET28aSEA. The target protein was induced by IPTG induction. The target protein with His Tag was purified by Ni NTAHisBindResin . Immunohistochemistry, immunofluorescence and flow cytometry were used to detect the anchoring effect on the cell membrane. To establish a melanoma model of C5 7BL 6 mice and observe the immunotherapeutic effects of the tumor vaccine prepared by transmembrane-type superantigen SEA fusion protein on tumor-bearing mice. Results Transmembrane-type superantigen fusion protein can be anchored on the tumor cell membrane. Tumor vaccine prepared by fusion protein can significantly inhibit tumor growth in tumor-bearing mice and prolong its survival. Conclusion Tumor vaccine prepared by transmembrane-type superantigen fusion protein has a strong anti-tumor effect and may be used as an effective new vaccine for the treatment and prevention of tumors.