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葡萄球菌肠毒素M和N的表达及生物活性分析

Expression and bioactivity analysis of Staphylococcal enterotoxin M and N.

作者信息

Pan Ying-qiu, Ding Ding, Li Dan-xi, Chen Shu-qing

机构信息

Department of Biopharmaceutics, Institute of Pharmacology & Toxicology and Biochemical Pharmaceutics, Zhejiang University, Hangzhou 310058, PR China.

出版信息

Protein Expr Purif. 2007 Dec;56(2):286-92. doi: 10.1016/j.pep.2007.08.005. Epub 2007 Aug 24.

Abstract

Staphylococcal enterotoxins (SEs) are powerful superantigens that stimulate non-specific T-cell proliferation produced by Staphylococcus aureus and draw considerable attention as ideal drugs for cancer therapy. The filtrate of S. aureus culture has been used as ampul named Staphylococcal enterotoxin C injection in clinic for 10 years in China and proved to be effective. The superantigen SEC claimed to be the only active component without certifiable evidences. For further investigations of the active components of this injection and establishment of foundations for the development of novel anti-cancer drugs, in this research we extracted total DNA from S. aureus (FRI 1230), cloned, expressed and purified recombinant proteins of Staphylococcal enterotoxin M and N (rSEM and rSEN). The MTT assay of the purified rSEM and rSEN demonstrated that their abilities of stimulating T cells and inhibiting the proliferation of K562-ADM cells and B16 cells were equivalent to that of purified SEC2 in vitro. These findings suggested that SEC was not the only active component of Staphylococcal enterotoxin C injection and the effective procedure of expression and purification may be useful for mass productions of these therapeutically important proteins.

摘要

葡萄球菌肠毒素(SEs)是一类强大的超抗原,可刺激由金黄色葡萄球菌产生的非特异性T细胞增殖,作为癌症治疗的理想药物备受关注。在中国,金黄色葡萄球菌培养滤液作为注射剂名为葡萄球菌肠毒素C注射液已在临床上使用了10年,并被证明是有效的。超抗原SEC被认为是唯一的活性成分,但并无确凿证据。为了进一步研究该注射液的活性成分,并为新型抗癌药物的开发奠定基础,在本研究中,我们从金黄色葡萄球菌(FRI 1230)中提取了总DNA,克隆、表达并纯化了葡萄球菌肠毒素M和N(rSEM和rSEN)的重组蛋白。纯化后的rSEM和rSEN的MTT试验表明,它们在体外刺激T细胞以及抑制K562-ADM细胞和B16细胞增殖的能力与纯化后的SEC2相当。这些发现表明,SEC并非葡萄球菌肠毒素C注射液的唯一活性成分,有效的表达和纯化方法可能有助于大规模生产这些具有重要治疗意义的蛋白质。

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