旨在探究非洲猪瘟病毒(African swine fever virus,ASFV)D1133L单克隆抗体对病毒复制的调控效应.在制备了 ASFV D1133L单克隆抗体的基础上,将ASFV和不同浓度的单克隆抗体同时接种于PAMs,通过红细胞吸附试验(HAD50)、实时荧光定量PCR(RT-qPCR)、蛋白质印迹(Western blot)和荧光观察分析D1133L单克隆抗体对病毒复制的影响.结果显示:不同浓度的单克隆抗体在不同的感染时间对ASFV的病毒效价(P<0.01),蛋白表达水平,基因转录水平(P<0.01),ASFV-GFP绿色荧光蛋白的表达水平(P<0.05)均有显著抑制,且这种抑制作用具有剂量依赖性.综上所述,D1133L单克隆抗体可显著抑制ASFV的复制,试验结果在ASFV药物靶点的选择上具有一定的参考意义.
The aim was to investigate the regulatory effect of a monoclonal antibody against D1133L protein of African swine fever virus(ASFV)during virus replication.Based on the preparation of monoclonal antibody against D1133L protein,ASFV and different concentrations of monoclonal antibodies were added simultaneously to PAMs,and the effect of monoclonal antibody against D1133L on virus replication was analyzed by 50%Hemadsorbing dose(HAD5o),real-time quantitative PCR(RT-qPCR),Western blot,and observation of fluorescence.Monoclonal antibodies at different concentrations significantly inhibited the viral titer(P<0.01),the expression level of proteins,the gene transcription level(P<0.01),and the expression level of ASFV-GFP green fluorescent protein(P<0.05)at different infection times in a dose-dependent manner.In summary,monoclonal antibody against D1133L significantly inhibited ASFV replication,the results of the trial are informative in the selection of drug targets for ASFV.