非洲猪瘟病毒D1133 L蛋白单克隆抗体抑制其复制

doi:10.11843/j.issn.0366-6964.2024.02.041

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (2): 854-859.doi: 10.11843/j.issn.0366-6964.2024.02.041

非洲猪瘟病毒D1133 L蛋白单克隆抗体抑制其复制

闫文倩^1,2, 侯景^1,2, 杨金柯^1,2, 郝雨^1,2, 杨行^1,2, 史喜绢^1,2, 张大俊^1,2, 别鑫恬^1,2, 陈国辉^1,2, 陈玲玲^1,2, 何路^1,2, 赵美玉^1,2, 赵思越^1,2, 郑海学^1,2*, 张克山^1,2*

1. 中国农业科学院兰州兽医研究所兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室, 兰州 730000;
2. 甘肃省病原生物学基础学科研究中心, 兰州 730046

收稿日期:2023-05-04 出版日期:2024-02-23 发布日期:2024-02-27
通讯作者: 郑海学,主要从事动物传染病学与流行病学研究,E-mail:zhenghaixue@caas.cn;张克山,主要从事兽医微生物及其分子生物学研究,E-mail:zks009@126.com
作者简介:闫文倩(1998-),女,山东滨州人,硕士生,主要从事兽医微生物及其分子生物学研究,E-mail:YWQ2373564886@163.com
基金资助:
国家重点研发计划(2021YFD1801300);2022甘肃省创新联合体-家畜重要疫病诊断研制与产业化(22ZD6NA012);中央高校基本科研业务费专项资金资助;"十四五"广东省农业科技创新十大主攻方向"揭榜挂帅"项目(2023SDZG02);中国农业科学院重大任务(CAAS-ZDRW202006-03);甘肃省科技重大专项(20ZD7NA006-2)

Monoclonal Antibody against D1133 L Protein of African Swine Fever Virus Inhibits Its Replication

YAN Wenqian^1,2, HOU Jing^1,2, YANG Jinke^1,2, HAO Yu^1,2, YANG Xing^1,2, SHI Xijuan^1,2, ZHANG Dajun^1,2, BIE Xintian^1,2, CHEN Guohui^1,2, CHEN Lingling^1,2, HE Lu^1,2, ZHAO Meiyu^1,2, ZHAO Siyue^1,2, ZHENG Haixue^1,2*, ZHANG Keshan^1,2*

1. State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730000, China;
2. Gansu Province Research Center for Basic Disciplines of Pathogen Biology, Lanzhou 730046, China

Received:2023-05-04 Online:2024-02-23 Published:2024-02-27

摘要/Abstract

摘要： 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。

关键词: 非洲猪瘟病毒, D1133 L蛋白, 单克隆抗体

Abstract: 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 (HAD₅₀), 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.

Key words: African swine fever virus, D1133 L protein, monoclonal antibody

中图分类号:

S852.659.1

闫文倩, 侯景, 杨金柯, 郝雨, 杨行, 史喜绢, 张大俊, 别鑫恬, 陈国辉, 陈玲玲, 何路, 赵美玉, 赵思越, 郑海学, 张克山. 非洲猪瘟病毒D1133 L蛋白单克隆抗体抑制其复制[J]. 畜牧兽医学报, 2024, 55(2): 854-859.

YAN Wenqian, HOU Jing, YANG Jinke, HAO Yu, YANG Xing, SHI Xijuan, ZHANG Dajun, BIE Xintian, CHEN Guohui, CHEN Lingling, HE Lu, ZHAO Meiyu, ZHAO Siyue, ZHENG Haixue, ZHANG Keshan. Monoclonal Antibody against D1133 L Protein of African Swine Fever Virus Inhibits Its Replication[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 854-859.

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非洲猪瘟病毒D1133 L蛋白单克隆抗体抑制其复制

Monoclonal Antibody against D1133 L Protein of African Swine Fever Virus Inhibits Its Replication

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