登革病毒单克隆抗体制备及其生物学性状分析  

Preparation and characterization of monoclonal antibody against dengue virus

作　　者：杨柳[1] 任瑞文[1] 张培[1] 管文升 陈颖龙[2] 朱明星[1] 陈月[1] 唐博恒[1] 

机构地区：[1]广州军区疾病预防控制中心,广东广州510507 [2]南方医科大学生物技术学院,广东广州510515

出　　处：《热带医学杂志》2015年第2期178-181,F0004,共5页Journal of Tropical Medicine

基　　金：全军医学科技"十二五"科研项目(CWS11C267);全军卫生和疾病防控应用性指令项目(13BJY20);广州市科技计划项目(2014J4100003)

摘　　要：目的筛选鉴定抗登革1~4型病毒单克隆抗体,并对其亚型、免疫学交叉反应及病毒中和能力进行初步分析。方法登革1~4型病毒乳鼠脑悬液混合后免疫BALB/c小鼠,经细胞融合后,免疫荧光法筛选抗登革病毒单克隆抗体,并在Hela细胞水平,病毒中和实验鉴定其对登革1型、2型病毒的中和能力,并详细分析所获中和抗体的亚型及免疫学交叉反应性。结果筛选获得抗登革病毒单克隆抗体41株,初步的中和实验结果表明,其中7株具登革1型病毒中和能力,13株具登革2型病毒中和能力,分别属于Ig G1、Ig G3、Ig G2a、Ig G2b、Ig M亚型,免疫荧光法交叉反应分析表明与登革1型病毒、登革2型病毒、登革3型病毒、登革4型病毒、乙型脑炎病毒、黄热病毒存在广泛免疫学交叉反应。结论筛选鉴定系列登革病毒中和抗体,为进一步探讨其中和作用的分子机制奠定了基础。Objective To prepare and characterize anti-dengue virus monoclonal antibody. Methods Female BALB /c mice of 6-8 weeks old were immunized with dengue virus type 1-4 infected neonatal mice brain suspensions. Hybridoma cells secreting monoclonal antibodies （McAbs） against dengue virus were screened using immunofluore scence method. Then their neutralizing capacity against dengue virus type 1 and 2 were further investigated. The isotyping and cross reaction capacity of the virus neutralizing antibodies were also investigated. Results 41 hybridoma cell lines named mAbl^41 were obtained, which had the property of secreting anti-dengue virus monoclonal antibody steadily.7 antibodies secreted by these hybridoma cell lines can neutralize dengue virus type 1 ,while 13 can neutralize dengue virus type 2. Isotyping analysis suggested these antibodies belong to IgG1, IgG3, IgG2a, IgG2b and IgM subtype, and show a widely immunology cross reaction capacity against dengue virus type 1-4,Japanese encephalitis virus and Yellow fever virus. Conclusion Series of neutralizing monoclonal antibodies against dengue virus has been developed successfully, which may lay a foundation for further study on the function of their antigens.

关 键 词：登革病毒 单克隆抗体 中和抗体 

分 类 号：R392.1[医药卫生—免疫学]