戊型肝炎病毒高通量中和效价评价模型的建立  

The Establishment of High-throughput Neutralization Titer Evaluation Model for Hepatitis E Virus(HEV)

作　　者：杨帆[1] 唐自闽 王思令 蔡薇[2] 温桂平[2] 纪文芳 余静菲 张可[1] 夏宁邵[1,2] 郑子峥[1] 

机构地区：[1]厦门大学分子疫苗学与分子诊断学国家重点实验室、国家传染病诊断试剂与疫苗工程技术研究中心、公共卫生学院,厦门361102 [2]厦门大学分子疫苗学与分子诊断学国家重点实验室、国家传染病诊断试剂与疫苗工程技术研究中心、生命科学院,厦门361102

出　　处：《病毒学报》2015年第1期1-6,共6页Chinese Journal of Virology

基　　金：国家基金(81373061);重大新药创制(2013ZX09101017);厦门市科技计划项目(3502Z20131001)

摘　　要：广泛有效的体外感染模型的缺乏限制了抗戊型肝炎病毒(Hepatitis E virus,HEV)抗体及血清定量化中和效价的评估,从而阻碍了对HEV相关抗体应答及免疫机制的深入研究。本研究首先通过在HEV低效复制细胞系HepG2肝癌细胞株上进行了HEV感染细胞后的连续监测,直到第13d到达病毒载量的检测下限,验证了该细胞系建立感染模型的可行性,进一步采用96孔多通道平行感染、核酸提取及实时荧光定量PCR对五株抗HEV的鼠源单克隆抗体及四位戊肝疫苗接种者的接种前、后血清样本进行了细胞中和试验。结果显示应用该模型能够实现对不同中和能力的单抗及接种疫苗前后的血清进行中和效价的定量化评估。表明本研究已成功建立了HEV体外高通量中和评价模型,同时也显示出该模型在HEV疫苗及抗体应答表位研究中所具有的潜在价值。The lack of effective in vitro infection model for hepatitis E virus （HEV） has greatly hindered the quantitative analysis of neutralizing titers of anti-HEV antibodies and human sera, thus impeding further studies of HEV-stimulated antibody responses and the immunological mechanisms. In order to improve this situation, the infection of HepG2 cells that are inefficient for HEV replication was continuously monitored until the viral load reached the limit of detection on day 13, the results of which confirmed the feasi- bility of using this cell line to establish the infection model. Then, neutralization assays of five anti-HEV murine monoclonal antibodies and serum samples collected from four HEV vaccine recipients （collected before and after vaccination） were performed by 96 multi-channel parallel infections, nucleic acid extraction, and qPCR. The results showed that the cell model can be applied for quantitative evaluation of the neutralizing capacity of different antibodies and antiserum samples from HEV vaccine recipients. In this study, we have successfully established a high-throughput in vitro HEV replication model, which will prove to be useful for the evaluation of HEV vaccines and studies of HEV epitopes.

关 键 词：阿霉素肾纤维化 三七注射液 转化生长因子-Β1 P38MAPK 

分 类 号：R373.9[医药卫生—病原生物学]