Hui Jing, Cao Yan, Xiao Fang, Zhang Jin, Li Hui, Hu Fengqing
School of Life Science, Liaoning University, Shenyang, 110036, China.
J Ind Microbiol Biotechnol. 2008 Sep;35(9):975-80. doi: 10.1007/s10295-008-0372-3. Epub 2008 May 28.
Staphylococcal enterotoxin C2 (SEC2) is one member of bacterial superantigens produced by Staphylococcus aureus. It can be attributed to its superantigenic activity to cross-link major histocompatibility complex class II molecules with T-cell receptors and activate a large number of resting T cells resulting in release of massive cytokines, which will produce significant tumor inhibition in vivo and in vitro. However, it could be not broadly applied to cure malignant tumors in clinic because of emetic activity of SEC2. The aim of this study was to inactivate emetic activity of SEC2 through site-directed mutagenesis. Cys93, Cys110 and His118 were selected as substitutional sites based on the functional sites responsible for emesis. The mutated proteins were used to determine Peripheral blood mononuclear cell proliferation activity and anti-tumor activity in vitro. Results showed that these mutated proteins efficiently stimulated T cell and exhibited the same tumor-inhibition effect as SEC2. It is possible to inactivate emetic activity of SEC2 through site-directed mutagenesis and provide satisfying agents for tumor treatment in clinic.
葡萄球菌肠毒素C2(SEC2)是金黄色葡萄球菌产生的细菌超抗原之一。它可归因于其超抗原活性,即能使主要组织相容性复合体II类分子与T细胞受体交联,并激活大量静息T细胞,导致大量细胞因子释放,从而在体内和体外产生显著的肿瘤抑制作用。然而,由于SEC2的催吐活性,它在临床上不能广泛应用于治疗恶性肿瘤。本研究的目的是通过定点诱变使SEC2的催吐活性失活。基于负责催吐的功能位点,选择Cys93、Cys110和His118作为替代位点。用突变蛋白测定外周血单个核细胞增殖活性和体外抗肿瘤活性。结果表明,这些突变蛋白能有效刺激T细胞,并表现出与SEC2相同的肿瘤抑制作用。通过定点诱变使SEC2的催吐活性失活并为临床肿瘤治疗提供满意的药物是可能的。