金黄色葡萄球菌肠毒素A在免疫应答中的作用

The Role of Staphylococcal Enterotoxin A in the Immune Response

目的获得重组表达、纯化金黄色葡萄球菌肠毒素A(SEA),并研究其免疫增强作用。方法利用基因工程法从金黄色葡萄球菌中克隆肠毒素A基因(sea),构建重组表达载体pET32a-sea,转化大肠杆菌(Rosetta)进行表达,并对其进行纯化;纯化后的SEA在白细胞介素2(IL-2)参与下,与半胱氨酸蛋白酶抑制剂C(CysC)进行动物免疫实验,检测其免疫增强效果。结果纯化后的SEA产量大约为40mg/L菌液,SDS-PAGE分析在27kD处有单一的目的条带;与对照组相比,SEA在IL-2的参与下与抗原共同免疫,抗体效价提高2.5倍。结论 SEA能够明显增强抗原的免疫应答效果。

Objective To recombine and purify the Staphylococcal enterotoxin A（SEA）gene and detect the immune activity of the recombinant SEA.Methods The Staphylococcal enterotoxin A（SEA）gene was cloned into the prokaryotic plasmid pET32a-sea by means of the genetic engineering method.The recombinant protein was expressed in E.coli Rosetta strains and purified by nickle affinity chromatography column.Interleukin-2（IL-2）and Cystatin C（Cys C）were used to detect the immune activity of the purified SEA.Results The SEA was expressed in E.coli Rosetta successfully and purified with a production rate of 40 mg/L.SDS-PAGE analysis showed a molecular weight of approximately 27 kDa for a subunit of the protein.The immune experiments showed that the SEA enhanced the immune activity by nearly 2.5 times.Conclusion SEA can significantly enhance the immune effect of antigen.