登革病毒prM抗体的中和作用及ADE作用研究  

Neutralizing Effect and Antibody Dependent Enhancement Activity of Anti-DENV prM Polyclonal Antibody

作　　者：司露露 郭晓兰[1] 郭鹏娟 王颖[1] 罗雅艳[1] 崔国辉[1] 方丹云[1] 周俊梅[1] 晏辉钧[1] 江丽芳[1] 

机构地区：[1]中山大学热带病防治研究教育部重点实验室//中山医学院微生物学教研室,广东广州510080 [2]海珠区疾病预防控制中心,广东广州510288

出　　处：《中山大学学报（医学科学版）》2016年第3期321-325,共5页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：NFSC-广东省联合基金(U1132002);国家自然科学基金(31270974);广东省科技计划项目(2013A020229004);广州市健康医疗协同创新重大专项(201508020263)

摘　　要：【目的】研究登革病毒(DENV)前膜蛋白(pr M)多克隆抗体对不同型别登革病毒的中和作用和抗体依赖性感染增强作用(ADE),为进一步阐明pr M蛋白在登革病毒致病与免疫过程中的重要作用提供依据。【方法】采用ELISA方法检测登革热患者血清中是否存在pr M抗体;用重组登革病毒2型(DENV-2)pr M蛋白免疫家兔,制备pr M蛋白多克隆抗体;采用空斑减数中和试验和流式细胞术检测pr M抗体对4种血清型登革病毒(DENV-1、DENV-2、DENV-3、DENV-4)的中和作用及ADE作用。【结果】登革热患者血清中存在特异性pr M抗体;pr M抗体对4种血清型登革病毒只有部分中和作用,但在一定浓度范围内对DENV1-4型均有明显的感染增强活性,且对异型DENV的感染增强作用强于同型,其中对登革病毒2型未成熟颗粒(im DENV-2)的感染增强作用最为明显。【结论】登革热患者血清存在pr M特异性抗体;pr M抗体对不同血清型DENV均只有较弱的中和活性,但却呈现出较强的ADE效应,证明登革病毒pr M抗体是一种感染增强性抗体。[Objective] To determine the neutralizing effect and antibody dependent enhancement （ADE） activity of anti-precursor-membrane protein （prM） antibody to all DENV serotypes, which will facilitate the understand of the important role of prM in the pathogenesis and immune course of DENV infection. [Methods]The level of anti-prM antibody in DENV-infected patients was detected by ELISA. The recombinant DENV-2 prM protein was used to immunize the rabbit to produce polyclonal antibody （pAb） against prM. The neutralizing effect of pAb was tested by plaque reduction neutralization test （PRNT）, and the ADE activity was tested by flow eytometry （FCM）. [ Results ] The prM-speeific antibody was found in sera of DENV- infected patients. The pAb against prM could not completely neutralize the infection of all four DENV serotypes, however, it was able to enhance the infectivity of DENVs. In addition, the enhancement to heterologous serotype was higher than homologous serotype, especially for immature particles. [ Conclusions ] Substantial level of prM antibody was found in sera of DENV- infected patients. The pAb against prM showed a weak neutralizing effect but a potent ADE activity to all four DENV serotypes. Together, these data suggest that anti-prM antibody had an infection-enhancing effect.

关 键 词：登革病毒 前膜蛋白(prM) 多克隆抗体 中和作用 抗体依赖性感染增强作用(ADE) 

分 类 号：R373.33[医药卫生—病原生物学]