登革病毒EDⅢ特异性单抗的体内外中和活性观察  

A murine monoclone antibody against envelope protein domain Ⅲ neutralizes dengue virus four serotypes in vitro and in vivo

作　　者：黄艳芬[1] 邓永强[2] 丘立文[1] 刘利东[1] 秦鄂德[2] 车小燕[1] 秦成峰[2] 

机构地区：[1]南方医科大学珠江医院检验医学部,广州510282 [2]军事医学科学院微生物流行病研究所,北京100071

出　　处：《军事医学》2013年第5期345-348,共4页Military Medical Sciences

基　　金：国家自然科学基金资助项目(U1132002;81101243;31270974)

摘　　要：目的鉴定1株能同时识别登革病毒Ⅰ～Ⅳ型(dengue virus serotypesⅠ-Ⅳ,DENV 1～4)的EDⅢ特异性单抗2D73的体内外中和活性。方法采用间接酶联免疫吸附实验(ELISA)和间接免疫荧光法(IFA)对单抗2D73的特性进行鉴定,并用基于酶联免疫斑点法的微中和实验(ELISPOT-MNT)和乳鼠保护实验观察单抗2D73对DENV 1～4的体内外中和活性。结果单抗2D73对4型DENV均具有较强的中和活性,其50%抑制浓度(IC50)分别为0.28、0.16、0.18和18.82μg/ml。乳鼠保护实验显示,该抗体对DENV 1～4同样具有较好的体内保护效果,与对照组相比,实验组小鼠发病时间明显延迟,生存率显著提高(P<0.05)。结论获得1株同时针对DENV1～4且具有体内外中和活性的单抗,该单抗可进一步用于DENV致病和免疫机制的研究以及抗病毒药物的研制。Objective To evaluate the neutralizing activities of a murine monoclonal antibody（mAb） 2D73 against dengue virus serotypesⅠ-Ⅳ（DENV1-4） in vitro and in vivo.Methods Indirect ELISA and immuno-fluorescence assay（IFA） were applied to identify specificity of mAb 2D73.Enzyme-linked immunospot micro-neutralizing test（ELISPOT-MNT） was performed to identify its neutralizing activities in vitro,and protective capacity was evaluated in a stringent intracranial challenge suckling mice model.Results All the four DENV serotypes were strongly neutralized by mAb 2D73 with 50% inhibitory concentrations（IC50） of 0.28,0.16,0.18 and 18.82 μg/ml to DENV1-4,respectively.The onset of pathogenesis was delayed and the survival rate was improved obviously（P0.05） in experimental group compared with control group.Conclusion MAb 2D73 against all the four DENV serotypes has strong neutralizing activities in vitro and protective efficacy in vivo,and might be used for the study of the pathogenic and immunologic mechanism of DENV and for the development of antiviral therapy.

关 键 词：登革病毒 包膜蛋白结构域Ⅲ 中和抗体 微中和实验 

分 类 号：R373.33[医药卫生—病原生物学]