非洲猪瘟病毒p30蛋白线性抗原表位的鉴定

doi:10.1016/j.jia.2023.04.012

Journal of Integrative Agriculture ›› 2023, Vol. 22 ›› Issue (6): 1945-1949.DOI: 10.1016/j.jia.2023.04.012

非洲猪瘟病毒p30蛋白线性抗原表位的鉴定

收稿日期:2023-01-06 修回日期:2023-04-15 接受日期:2023-03-15 出版日期:2023-06-20 发布日期:2023-03-15

Identification of two novel linear epitopes on the p30 protein of African swine fever virus

YU Si-hui^1*, SHAN Zhao-meng^1*, YANG Jing-jing¹, LIU Yi-ning¹, WU Chang-de¹, ZHANG Zhen-jiang², ZHU Yuan-mao², MENG Bo¹, ZHAN Jia-xing¹, WEN Xue-xia^1#, ZHANG Ying^1#

¹Key Laboratory of Livestock Infectious Diseases of Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, P.R.China
²State Key Laboratory of Veterinary Biotechnology, National High Containment Facilities for Animal Diseases Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.China

Received:2023-01-06 Revised:2023-04-15 Accepted:2023-03-15 Online:2023-06-20 Published:2023-03-15
About author:#Correspondence WEN Xue-xia, Tel: +86-24-88487156, E-mail: wenxuexia123@syau.edu.cn; ZHANG Ying, Tel: +86-24-88487156, E-mail: yingzhang18@syau.edu.cn * These authors contributed equally to this study.
Supported by:
This study was supported by the National Natural Science Foundation of China (31902258), the Scientific Research Staring Foundation for the Doctors from the Science and Technology Department of Liaoning Province, China (2019-BS-204), and the Major Project of Science and Technology of Liaoning Province (2020JH1/10200003).

摘要/Abstract

摘要：

非洲猪瘟（ASF）的持续流行严重威胁全球养猪业的健康发展。该病目前缺乏可应用于临床的疫苗和药物，因此，早期的精准检测对于ASF的有效防控至关重要。p30蛋白在病毒感染早期表达，免疫原性较好且诱导产生抗体水平较高，这些特性使其成为较为理想的非洲猪瘟病毒（ASFV）检测靶点之一。本研究以原核表达的p30蛋白为免疫原，制备了两株可稳定分泌针对ASFV p30蛋白的单克隆抗体（mAb）细胞株，分别命名为4F2和7F3。利用Western blotting和间接免疫荧光实验鉴定两株单克隆抗体均能识别病毒感染。亚型鉴定结果显示，4F2和7F3的重链分别为IgG1和IgG2b，轻链均为κ链。随后，通过原核表达一系列截短p30蛋白鉴定4F2和7F3识别的抗原表位。经5轮截短和Western blotting实验鉴定，最终确定4F2和7F3识别的最短抗原表位分别为¹¹⁸SFETL¹²²和⁷⁶EHQAQEEWNMILHV⁸⁹。在此基础上，进一步分析两个抗原表位在不同毒株p30蛋白序列中的保守性。序列比对结果表明，GenBank数据库中所有p30蛋白序列均包含¹¹⁸SFETL¹²² ，而另一抗原表位⁷⁶EHQAQEEWNMILHV⁸⁹则在非洲以外地区分离的毒株中保守。本研究制备的两株单克隆抗体及鉴定的抗原表位可为检测方法的开发提供材料和基础，同时也可为病毒致病机理和免疫机制等基础研究提供工具。

YU Si-hui, SHAN Zhao-meng, YANG Jing-jing, LIU Yi-ning, WU Chang-de, ZHANG Zhen-jiang, ZHU Yuan-mao, MENG Bo, ZHAN Jia-xing, WEN Xue-xia, ZHANG Ying. 非洲猪瘟病毒p30蛋白线性抗原表位的鉴定[J]. Journal of Integrative Agriculture, 2023, 22(6): 1945-1949.

YU Si-hui, SHAN Zhao-meng, YANG Jing-jing, LIU Yi-ning, WU Chang-de, ZHANG Zhen-jiang, ZHU Yuan-mao, MENG Bo, ZHAN Jia-xing, WEN Xue-xia, ZHANG Ying. Identification of two novel linear epitopes on the p30 protein of African swine fever virus[J]. Journal of Integrative Agriculture, 2023, 22(6): 1945-1949.

参考文献

Afonso C L, Alcaraz C, Brun A, Sussman M D, Onisk D V, Escribano J M, Rock D L. 1992. Characterization of p30, a highly antigenic membrane and secreted protein of African swine fever virus. Virology, 189, 368–373.

Barderas M G, Rodriguez F, Gomez-Puertas P, Aviles M, Beitia F, Alonso C, Escribano J M. 2001. Antigenic and immunogenic properties of a chimera of two immunodominant African swine fever virus proteins. Archives of Virology, 146, 1681–1691.

Bishop R P, Fleischauer C, De Villiers E P, Okoth E A, Arias M, Gallardo C, Upton C. 2015. Comparative analysis of the complete genome sequences of Kenyan African swine fever virus isolates within p72 genotypes IX and X. Virus Genes, 50, 303–309.

Bisimwa P N, Ongus J R, Steinaa L, Bisimwa E B, Bochere E, Machuka E M, Entfellner J D, Okoth E, Pelle R. 2021. The first complete genome sequence of the African swine fever virus genotype X and serogroup 7 isolated in domestic pigs from the Democratic Republic of Congo. Virology Journal, 18, 23.

Chapman D A, Darby A C, Da Silva M, Upton C, Radford A D, Dixon L K. 2011. Genomic analysis of highly virulent Georgia 2007/1 isolate of African swine fever virus. Emerging Infectious Diseases, 17, 599–605.

Chapman D A G, Tcherepanov V, Upton C, Dixon L K. 2008. Comparison of the genome sequences of non-pathogenic and pathogenic African swine fever virus isolates. Journal of General Virology, 89, 397–408.

Costard S, Wieland B, De Glanville W, Jori F, Rowlands R, Vosloo W, Roger F, Pfeiffer D U, Dixon L K. 2009. African swine fever: How can global spread be prevented? Philosophical Transactions of the Royal Society of London (B: Biological Sciences), 364, 2683–2696.

Gallardo C, Okoth E, Pelayo V, Anchuelo R, Martin E, Simon A, Llorente A, Nieto R, Soler A, Martin R, Arias M, Bishop R P. 2011. African swine fever viruses with two different genotypes, both of which occur in domestic pigs, are associated with ticks and adult warthogs, respectively, at a single geographical site. Journal of General Virology, 92, 432–444.

Garigliany M, Desmecht D, Tignon M, Cassart D, Lesenfant C, Paternostre J, Volpe R, Cay A B, Van Den Berg T, Linden A. 2019. Phylogeographic analysis of African swine fever virus, Western Europe, 2018. Emerging Infectious Diseases, 25, 184–186.

Gimenez-Lirola L G, Mur L, Rivera B, Mogler M, Sun Y, Lizano S, Goodell C, Harris D L, Rowland R R, Gallardo C, Sanchez-Vizcaino J M, Zimmerman J. 2016. Detection of African swine fever virus antibodies in serum and oral fluid specimens using a recombinant protein 30 (p30) dual matrix indirect ELISA. PLoS ONE, 11, e0161230.

Gomez-Puertas P, Rodriguez F, Oviedo J M, Brun A, Alonso C, Escribano J M. 1998. The African swine fever virus proteins p54 and p30 are involved in two distinct steps of virus attachment and both contribute to the antibody-mediated protective immune response. Virology, 243, 461–471.

Hakizimana J N, Ntirandekura J B, Yona C, Nyabongo L, Kamwendo G, Chulu J L C, Ntakirutimana D, Kamana O, Nauwynck H, Misinzo G. 2021. Complete genome analysis of African swine fever virus responsible for outbreaks in domestic pigs in 2018 in Burundi and 2019 in Malawi. Tropical Animal Health and Production, 53, 438.

Ivanov V, Efremov E E, Novikov B V, Balyshev V M, Tsibanov S, Kalinovsky T, Kolbasov D V, Niedzwiecki A, Rath M. 2011. Vaccination with viral protein-mimicking peptides postpones mortality in domestic pigs infected by African swine fever virus. Molecular Medicine Reports, 4, 395–401.

Jiang C G, Sun Y, Fan Z, Xin A, Feng X N, Wei H, Zhang X F, Zhao D M, Bu Z G, He X J. 2021. Viricidal activity of several disinfectants against African swine fever virus. Journal of Integrative Agriculture, 20, 3084–3088.

Kolbasov D, Titov I, Tsybanov S, Gogin A, Malogolovkin A. 2018. African swine fever virus, Siberia, Russia, 2017. Emerging Infectious Diseases, 24, 796–798.

Linden A, Licoppe A, Volpe R, Paternostre J, Lesenfants C, Cassart D, Garigliany M, Tignon M, Van Den Berg T, Desmecht D, Cay A B. 2019. Summer 2018: African swine fever virus hits North-Western Europe. Transboundary and Emerging Diseases, 66, 54–55.

Mur L, Atzeni M, Martinez-Lopez B, Feliziani F, Rolesu S, Sanchez-Vizcaino J M. 2016. Thirty-five-year presence of African swine fever in Sardinia: History, evolution and risk factors for disease maintenance. Transboundary and Emerging Diseases, 63, e165–e177.

Murgia M V, Mogler M, Certoma A, Green D, Monaghan P, Williams D T, Rowland R R, Gaudreault N N. 2019. Evaluation of an African swine fever (ASF) vaccine strategy incorporating priming with an alphavirus-expressed antigen followed by boosting with attenuated ASF virus. Archives of Virology, 164, 359–370.

Olesen A S, Lohse L, Dalgaard M D, Wozniakowski G, Belsham G J, Botner A, Rasmussen T B. 2018. Complete genome sequence of an African swine fever virus (ASFV POL/2015/Podlaskie) determined directly from pig erythrocyte-associated nucleic acid. Journal of Virological Methods, 261, 14–16.

Petrovan V, Yuan F, Li Y, Shang P, Murgia M V, Misra S, Rowland R R, Fang Y. 2019. Development and characterization of monoclonal antibodies against p30 protein of African swine fever virus. Virus Research, 269, 197632.

Rowlands R J, Michaud V, Heath L, Hutchings G, Oura C, Vosloo W, Dwarka R, Onashvili T, Albina E, Dixon L K. 2008. African swine fever virus isolate, Georgia, 2007. Emerging Infectious Diseases, 14, 1870–1874.

Simulundu E, Lubaba C H, Van Heerden J, Kajihara M, Mataa L, Chambaro H M, Sinkala Y, Munjita S M, Munang’andu H M, Nalubamba K S, Samui K, Pandey G S, Takada A, Mweene A S. 2017. The epidemiology of African swine fever in “nonendemic” regions of Zambia (1989–2015): implications for disease prevention and control. Viruses, 9, 236.

Smietanka K, Wozniakowski G, Kozak E, Niemczuk K, Fraczyk M, Bocian L, Kowalczyk A, Pejsak Z. 2016. African swine fever epidemic, Poland, 2014–2015. Emerging Infectious Diseases, 22, 1201–1207.

Vita R, Mahajan S, Overton J A, Dhanda S K, Martini S, Cantrell J R, Wheeler D K, Sette A, Peters B. 2019. The Immune Epitope Database (IEDB): 2018 update. Nucleic Acids Research, 47, D339–D343.

Wang J, Shi X J, Sun H W, Chen H J. 2020. Insights into African swine fever virus immunoevasion strategies. Journal of Integrative Agriculture, 19, 11–22.

Wang P F, Wang M, Shi Z B, Sun Z Z, Wei L L, Liu Z S, Wang S D, He X J, Wang J F. 2022. Development of a recombinant pB602L-based indirect ELISA assay for detecting antibodies against African swine fever virus in pigs. Journal of Integrative Agriculture, 21, 819–825.

Wen X, He X, Zhang X, Zhang X, Liu L, Guan Y, Zhang Y, Bu Z. 2019. Genome sequences derived from pig and dried blood pig feed samples provide important insights into the transmission of African swine fever virus in China in 2018. Emerging Microbes and Infections, 8, 303–306.

Wu P, Lowe A D, Rodriguez Y Y, Murgia M V, Dodd K A, Rowland R R, Jia W. 2020. Antigenic regions of African swine fever virus phosphoprotein p30. Transboundary and Emerging Diseases, 67, 1942–1953.

Zhang X, Liu X, Wu X, Ren W, Zou Y, Xia X, Sun H. 2021. A colloidal gold test strip assay for the detection of African swine fever virus based on two monoclonal antibodies against p30. Archives of Virology, 166, 871–879.

Zhao D, Liu R, Zhang X, Li F, Wang J, Zhang J, Liu X, Wang L, Zhang J, Wu X, Guan Y, Chen W, Wang X, He X, Bu Z. 2019. Replication and virulence in pigs of the first African swine fever virus isolated in China. Emerging Microbes and Infections, 8, 438–447.

Zhou G, Shi Z, Luo J, Cao L, Yang B, Wan Y, Wang L, Song R, Ma Y, Tian H, Zheng H. 2022. Preparation and epitope mapping of monoclonal antibodies against African swine fever virus p30 protein. Applied Microbiology and Biotechnology, 106, 1199–1210.

Zhou X, Li N, Luo Y, Liu Y, Miao F, Chen T, Zhang S, Cao P, Li X, Tian K, Qiu H J, Hu R. 2018. Emergence of African swine fever in China, 2018. Transboundary and Emerging Diseases, 65, 1482–1484.

非洲猪瘟病毒p30蛋白线性抗原表位的鉴定

Identification of two novel linear epitopes on the p30 protein of African swine fever virus

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