Expression and bioactivity analysis of Staphylococcal enterotoxin M and N.
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.
Pan
Ying-qiu
YQ
Department of Biopharmaceutics, Institute of Pharmacology & Toxicology and Biochemical Pharmaceutics, Zhejiang University, Hangzhou 310058, PR China.
Ding
Ding
D
Li
Dan-xi
DX
Chen
Shu-qing
SQ
D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D004768:Enterotoxins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D006801:Humans; D051379:Mice; D008813:Mice, Inbred ICR; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D013211:Staphylococcus aureus / Q000235:genetics / Q000276:immunology*; D018089:Superantigens / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology; D014407:Tumor Cells, Cultured
D000942:Antigens, Bacterial; D004768:Enterotoxins; D011993:Recombinant Fusion Proteins; D018089:Superantigens