抗金黄色葡萄球菌肠毒素B保护性中和单克隆抗体的筛选及鉴定  

Sereening and characterization of neutralizing monoclonal antibodies against Staphylococcal enterotoxin B

作　　者：康延申[1,2] 王华菁[2] 杨海鸥[3] 谈文龙[3] 张波[2] 魏于全[1] 戴建新[2] 

机构地区：[1]四川大学国家生物治疗重点实验室,成都610065 [2]第二军医大学肿瘤研究所,抗体药物国家工程研究中心,上海200433 [3]上海交通大学医学院,上海200025

出　　处：《现代免疫学》2010年第4期298-302,共5页Current Immunology

摘　　要：金黄色葡萄球菌肠毒素B(SEB)属于热毒素超抗原家族。它通过激活T细胞,使T细胞大量增殖并释放大量炎症细胞因子,从而导致中毒性休克综合征及死亡。为了筛选获得抗SEB的中和保护性单克隆抗体,我们首先克隆了SEB基因,表达纯化获得GST-SEB融合蛋白;将之免疫BALB/c小鼠后,利用杂交瘤技术结合ELISA检测,筛选获得10株抗SEB单克隆抗体杂交瘤细胞株;ELISA和Western blot鉴定其分泌的抗体能与SEB特异结合;抗体亚型检测均为IgG1/k;体外实验证实,10株抗体中有6株(1A5、4A3、3E2、3C1、1D1、2G9)能够有效阻断SEB诱导的人外周血单核细胞(PBMC)的激活,具有中和保护作用。本研究为进一步获得抗SEB的人源化单克隆中和性抗体奠定了基础。Staphylococcal enterotoxin B （SEB） is a member of a family of toxins known as pyrogenic toxin superantigens （PTSAgs） that activate a large number of T-cells by cross-linking MHC class Ⅱ molecules with T-cell receptors in a V beta-restricted fashion,and cause massive production of inflammatory cytokines,leading to toxic shock syndrome and death. In order to identify and characterize monoclonal antibodies （mAbs） with the neutralize against SEB,SEB gene was cloned,and the fusion protein GST-SEB. After immunization of BALB/c mice with SEB,10 hybridoma cell lines secreting mAbs against SEB were obtained by hybridoma technique combined with ELISA. Their ability to secrete specific antibody and the specific binding power with SEB was confirmed by ELISA and Western blot. The subtypes of the mAbs were all proved to be IgG1/k. Through in vitro experimentation,6 of 10 mAbs （1A5,4A3,3E2,3C1,1D1,2G9）were confirmed to have the ability to neutralize SEB and to block the activation of human PBMC induced by SEB. This study provides the evidence that these mAbs have the potential to be developed to humanized neutralization mAbs against SEB.

关 键 词：单克隆抗体 金黄色葡萄球菌肠毒素B 中和保护 

分 类 号：R392.12[医药卫生—免疫学]