{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T22:43:59Z","timestamp":1791499439959,"version":"4.3.4"},"reference-count":62,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,2,22]],"date-time":"2021-02-22T00:00:00Z","timestamp":1613952000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:p>Porcine reproductive and respiratory syndrome (PRRS) is a devastating disease which impacts the pig industry worldwide. The disease is caused by PRRS viruses (PRRSV-1 and -2) which leads to abortions and other forms of reproductive failure in sows and severe respiratory disease in growing pigs. Current PRRSV vaccines provide limited protection; only providing complete protection against closely related strains. The development of improved PRRSV vaccines would benefit from an increased understanding of epitopes relevant to protection, including those recognized by antibodies which possess the ability to neutralize distantly related strains. In this work, a reverse vaccinology approach was taken; starting first with pigs known to have a broadly neutralizing antibody response and then investigating the responsible B cells\/antibodies through the isolation of PRRSV neutralizing monoclonal antibodies (mAbs). PBMCs were harvested from pigs sequentially exposed to a modified-live PRRSV-2 vaccine as well as divergent PRRSV-2 field isolates. Memory B cells were immortalized and a total of 5 PRRSV-specific B-cell populations were isolated. All identified PRRSV-specific antibodies were found to be broadly binding to all PRRSV-2 isolates tested, but not PRRSV-1 isolates. Antibodies against GP5 protein, commonly thought to possess a dominant PRRSV neutralizing epitope, were found to be highly abundant, as four out of five B cells populations were GP5 specific. One of the GP5-specific mAbs was shown to be neutralizing but this was only observed against homologous and not heterologous PRRSV strains. Further investigation of these antibodies, and others, may lead to the elucidation of conserved neutralizing epitopes that can be exploited for improved vaccine design and lays the groundwork for the study of broadly neutralizing antibodies against other porcine pathogens.<\/jats:p>","DOI":"10.3389\/fimmu.2021.638493","type":"journal-article","created":{"date-parts":[[2021,2,22]],"date-time":"2021-02-22T05:03:36Z","timestamp":1613970216000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":23,"title":["Isolation of Porcine Reproductive and Respiratory Syndrome Virus GP5-Specific, Neutralizing Monoclonal Antibodies From Hyperimmune Sows"],"prefix":"10.3389","volume":"12","author":[{"given":"Jordan E.","family":"Young","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cheryl M. T.","family":"Dvorak","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simon P.","family":"Graham","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael P.","family":"Murtaugh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,2,22]]},"reference":[{"key":"B1","doi-asserted-by":"crossref","DOI":"10.1002\/9781119350927","volume-title":"Diseases of Swine","author":"Zimmerman","year":"2019"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1016\/S0378-1135(02)00013-5","article-title":"Characterization of the carrier state in porcine reproductive and respiratory syndrome virus infection","volume":"86","author":"Horter","year":"2002","journal-title":"Vet Microbiol"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.74.22.10834-10837.2000","article-title":"Porcine reproductive and respiratory syndrome virus: description of persistence in individual pigs upon experimental infection","volume":"74","author":"Allende","year":"2000","journal-title":"J Virol"},{"key":"B4","doi-asserted-by":"publisher","first-page":"72","DOI":"10.31274\/ans_air-180814-28","article-title":"Assessment of the economic impact of porcine reproductive and respiratory syndrome virus on United States pork producers","volume":"21","author":"Holtkamp","year":"2013","journal-title":"J Swine Health Prod"},{"key":"B5","doi-asserted-by":"publisher","first-page":"16","DOI":"10.1016\/j.prevetmed.2017.04.006","article-title":"Cost of porcine reproductive and respiratory syndrome virus at individual farm level - An economic disease model","volume":"142","author":"Nathues","year":"2017","journal-title":"Prev Vet Med"},{"key":"B6","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.vetimm.2015.07.003","article-title":"Innate and adaptive immunity against Porcine Reproductive and Respiratory Syndrome Virus","volume":"167","author":"Loving","year":"2015","journal-title":"Vet Immunol Immunopathol"},{"key":"B7","doi-asserted-by":"publisher","first-page":"148","DOI":"10.3390\/v9060148","article-title":"Mechanisms of Adaptive Immunity to Porcine Reproductive and Respiratory Syndrome Virus","volume":"9","author":"Rahe","year":"2017","journal-title":"Viruses"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2017.04.016","article-title":"GP5 of porcine reproductive and respiratory syndrome virus (PRRSV) as a target for homologous and broadly neutralizing antibodies","volume":"209","author":"Popescu","year":"2017","journal-title":"Vet Microbiol"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-78-8-1867","article-title":"Monoclonal antibodies to the ORF5 product of porcine reproductive and respiratory syndrome virus define linear neutralizing determinants","volume":"78","author":"Pirzadeh","year":"1997","journal-title":"J Gen Virol"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1007\/s007050070079","article-title":"Categorization of North American porcine reproductive and respiratory syndrome viruses: epitopic profiles of the N, M, GP5 and GP3 proteins and susceptibility to neutralization","volume":"145","author":"Yang","year":"2000","journal-title":"Arch Virol"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.76.9.4241-4250.2002","article-title":"Identification of neutralizing and nonneutralizing epitopes in the porcine reproductive and respiratory syndrome virus GP5 ectodomain","volume":"76","author":"Ostrowski","year":"2002","journal-title":"J Virol"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-002-0887-2","article-title":"The primary neutralization epitope of porcine respiratory and reproductive syndrome virus strain VR-2332 is located in the middle of the GP5 ectodomain","volume":"147","author":"Plagemann","year":"2002","journal-title":"Arch Virol"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.80402-0","article-title":"Significance of the oligosaccharides of the porcine reproductive and respiratory syndrome virus glycoproteins GP2a and GP5 for infectious virus production","volume":"85","author":"Wissink","year":"2004","journal-title":"J Gen Virol"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1007\/s11262-007-0131-y","article-title":"Influence of porcine reproductive and respiratory syndrome virus GP5 glycoprotein N-linked glycans on immune responses in mice","volume":"35","author":"Jiang","year":"2007","journal-title":"Virus Genes"},{"key":"B15","doi-asserted-by":"publisher","first-page":"10","DOI":"10.1016\/j.vetmic.2012.08.005","article-title":"Significance of genetic variation of PRRSV ORF5 in virus neutralization and molecular determinants corresponding to cross neutralization among PRRS viruses","volume":"162","author":"Kim","year":"2013","journal-title":"Vet Microbiol"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2011.04.071","article-title":"Characterization of antigenic regions in the porcine reproductive and respiratory syndrome virus by the use of peptide-specific serum antibodies","volume":"29","author":"Vanhee","year":"2011","journal-title":"Vaccine"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2011.11.023","article-title":"Baculovirus as a PRRSV and PCV2 bivalent vaccine vector: baculovirus virions displaying simultaneously GP5 glycoprotein of PRRSV and capsid protein of PCV2","volume":"179","author":"Xu","year":"2012","journal-title":"J Virol Methods"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1007\/s11259-010-9364-7","article-title":"Immune responses in pigs induced by recombinant canine adenovirus 2 expressing the glycoprotein 5 of porcine reproductive and respiratory syndrome virus","volume":"34","author":"Zhou","year":"2010","journal-title":"Vet Res Commun"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2012.06.018","article-title":"Highly pathogenic porcine reproductive and respiratory syndrome virus GP5 B antigenic region is not a neutralizing antigenic region","volume":"159","author":"Leng","year":"2012","journal-title":"Vet Microbiol"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2012.08.026","article-title":"Dissociation of porcine reproductive and respiratory syndrome virus neutralization from antibodies specific to major envelope protein surface epitopes","volume":"433","author":"Li","year":"2012","journal-title":"Virology"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01774-09","article-title":"The minor envelope glycoproteins GP2a and GP4 of porcine reproductive and respiratory syndrome virus interact with the receptor CD163","volume":"84","author":"Das","year":"2010","journal-title":"J Virol"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00513-07","article-title":"CD163 expression confers susceptibility to porcine reproductive and respiratory syndrome viruses","volume":"81","author":"Calvert","year":"2007","journal-title":"J Virol"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.06836-11","article-title":"Arterivirus minor envelope proteins are a major determinant of viral tropism in cell culture","volume":"86","author":"Tian","year":"2012","journal-title":"J Virol"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-009-0582-7","article-title":"GP4 of porcine reproductive and respiratory syndrome virus contains a neutralizing epitope that is susceptible to immunoselection in vitro","volume":"155","author":"Costers","year":"2010","journal-title":"Arch Virol"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2010.08.026","article-title":"GP4-specific neutralizing antibodies might be a driving force in PRRSV evolution","volume":"154","author":"Costers","year":"2010","journal-title":"Virus Res"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1089\/vim.2010.0025","article-title":"A variable region in GP4 of European-type porcine reproductive and respiratory syndrome virus induces neutralizing antibodies against homologous but not heterologous virus strains","volume":"23","author":"Vanhee","year":"2010","journal-title":"Viral Immunol"},{"key":"B27","doi-asserted-by":"publisher","first-page":"96","DOI":"10.1016\/j.virol.2012.09.005","article-title":"DNA shuffling of the GP3 genes of porcine reproductive and respiratory syndrome virus (PRRSV) produces a chimeric virus with an improved cross-neutralizing ability against a heterologous PRRSV strain","volume":"434","author":"Zhou","year":"2012","journal-title":"Virology"},{"key":"B28","doi-asserted-by":"publisher","first-page":"695","DOI":"10.1007\/s11262-007-0143-7","article-title":"Analysis of immunogenicity of minor envelope protein GP3 of porcine reproductive and respiratory syndrome virus in mice","volume":"35","author":"Jiang","year":"2007","journal-title":"Virus Genes"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1089\/vim.2006.19.285","article-title":"Neutralizing antibody formation in swine infected with seven strains of porcine reproductive and respiratory syndrome virus as measured by indirect ELISA with peptides containing the GP5 neutralization epitope","volume":"19","author":"Plagemann","year":"2006","journal-title":"Viral Immunol"},{"key":"B30","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1006\/viro.2002.1612","article-title":"Passive transfer of virus-specific antibodies confers protection against reproductive failure induced by a virulent strain of porcine reproductive and respiratory syndrome virus and establishes sterilizing immunity","volume":"302","author":"Osorio","year":"2002","journal-title":"Virology"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2011.07.076","article-title":"Porcine Reproductive and Respiratory Syndrome Virus isolates differ in their susceptibility to neutralization","volume":"29","author":"Mart\u00ednez-Lobo","year":"2011","journal-title":"Vaccine"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.03287-14","article-title":"A single amino Acid deletion in the matrix protein of porcine reproductive and respiratory syndrome virus confers resistance to a polyclonal Swine antibody with broadly neutralizing activity","volume":"89","author":"Trible","year":"2015","journal-title":"J Virol"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2009.04.022","article-title":"Challenges for porcine reproductive and respiratory syndrome virus (PRRSV) vaccinology","volume":"27","author":"Kimman","year":"2009","journal-title":"Vaccine"},{"key":"B34","doi-asserted-by":"publisher","first-page":"56","DOI":"10.1016\/j.virusres.2015.03.016","article-title":"Broadly neutralizing antibodies against the rapidly evolving porcine reproductive and respiratory syndrome virus","volume":"203","author":"Robinson","year":"2015","journal-title":"Virus Res"},{"key":"B35","doi-asserted-by":"publisher","first-page":"416727","DOI":"10.1155\/2014\/416727","article-title":"Comparison of host immune responses to homologous and heterologous type II porcine reproductive and respiratory syndrome virus (PRRSV) challenge in vaccinated and unvaccinated pigs","volume":"2014","author":"Li","year":"2014","journal-title":"BioMed Res Int"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1016\/j.theriogenology.2006.04.043","article-title":"Reproductive performance of gilts following vaccination and subsequent heterologous challenge with European strains of porcine reproductive and respiratory syndrome virus","volume":"66","author":"Scortti","year":"2006","journal-title":"Theriogenology"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.00572","article-title":"Establishment of Systems to Enable Isolation of Porcine Monoclonal Antibodies Broadly Neutralizing the Porcine Reproductive and Respiratory Syndrome Virus","volume":"10","author":"Goldeck","year":"2019","journal-title":"Front Immunol"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2010.04.001","article-title":"A simple method for Alexa Fluor dye labelling of dengue virus","volume":"167","author":"Zhang","year":"2010","journal-title":"J Virol Methods"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1093\/molbev\/msy096","article-title":"MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms","volume":"35","author":"Kumar","year":"2018","journal-title":"Mol Biol Evol"},{"key":"B40","doi-asserted-by":"publisher","first-page":"e0171171","DOI":"10.1371\/journal.pone.0171171","article-title":"Interleukin-21 Drives Proliferation and Differentiation of Porcine Memory B Cells into Antibody Secreting Cells","volume":"12","author":"Rahe","year":"2017","journal-title":"PloS One"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1038\/nm.2071","article-title":"Generation of stable monoclonal antibody-producing B cell receptor-positive human memory B cells by genetic programming","volume":"16","author":"Kwakkenbos","year":"2010","journal-title":"Nat Med"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1128\/CVI.00483-08","article-title":"Antibody response to porcine reproductive and respiratory syndrome virus (PRRSV) nonstructural proteins and implications for diagnostic detection and differentiation of PRRSV types I and II","volume":"16","author":"Brown","year":"2009","journal-title":"Clin Vaccine Immunol"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01023-07","article-title":"Antigen-specific B-cell responses to porcine reproductive and respiratory syndrome virus infection","volume":"82","author":"Mulupuri","year":"2008","journal-title":"J Virol"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2008.08.002","article-title":"Immune response against porcine reproductive and respiratory syndrome virus during acute and chronic infection","volume":"126","author":"Molina","year":"2008","journal-title":"Vet Immunol Immunopathol"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.82587-0","article-title":"Cross-reactive antibody responses to nsp1 and nsp2 of Porcine reproductive and respiratory syndrome virus","volume":"88","author":"Johnson","year":"2007","journal-title":"J Gen Virol"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.73.3.2016-2026.1999","article-title":"Open reading frame 1a-encoded subunits of the arterivirus replicase induce endoplasmic reticulum-derived double-membrane vesicles which carry the viral replication complex","volume":"73","author":"Pedersen","year":"1999","journal-title":"J Virol"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.72.8.6689-6698.1998","article-title":"ORF1a-encoded replicase subunits are involved in the membrane association of the arterivirus replication complex","volume":"72","author":"van der Meer","year":"1998","journal-title":"J Virol"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.70.10.6625-6633.1996","article-title":"Processing of the equine arteritis virus replicase ORF1b protein: identification of cleavage products containing the putative viral polymerase and helicase domains","volume":"70","author":"van Dinten","year":"1996","journal-title":"J Virol"},{"key":"B49","doi-asserted-by":"publisher","first-page":"3994","DOI":"10.1128\/JVI.80.8.3994-4004.2006","article-title":"Influence of N-linked glycosylation of porcine reproductive and respiratory syndrome virus GP5 on virus infectivity, antigenicity, and ability to induce neutralizing antibodies","volume":"80","author":"Ansari","year":"2006","journal-title":"J Virol"},{"key":"B50","doi-asserted-by":"publisher","first-page":"294","DOI":"10.1089\/vim.2006.19.294","article-title":"Neutralizing antibody responses of pigs infected with natural GP5 N-glycan mutants of porcine reproductive and respiratory syndrome virus","volume":"19","author":"Faaberg","year":"2006","journal-title":"Viral Immunol"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1292\/jvms.63.567","article-title":"Polymerase chain reaction-based restriction fragment length polymorphism pattern of porcine reproductive and respiratory syndrome virus directly from lung tissues without virus isolation in Korea","volume":"63","author":"Cheon","year":"2001","journal-title":"J Vet Med Sci"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1051\/vetres:2000108","article-title":"A brief review of procedures and potential problems associated with the diagnosis of porcine reproductive and respiratory syndrome","volume":"31","author":"Mengeling","year":"2000","journal-title":"Vet Res"},{"key":"B53","doi-asserted-by":"publisher","DOI":"10.1177\/104063870201400415","article-title":"Restriction fragment length polymorphism of porcine reproductive and respiratory syndrome viruses recovered from Ontario farms, 1998-2000","volume":"14","author":"Cai","year":"2002","journal-title":"J Vet Diagn Invest"},{"key":"B54","doi-asserted-by":"publisher","DOI":"10.1177\/104063879400600402","article-title":"Serum immune responses to the proteins of porcine reproductive and respiratory syndrome (PRRS) virus","volume":"6","author":"Nelson","year":"1994","journal-title":"J Vet Diagn Invest"},{"key":"B55","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.80675-0","article-title":"Analysis of porcine reproductive and respiratory syndrome virus attachment and internalization: distinctive roles for heparan sulphate and sialoadhesin","volume":"86","author":"Delputte","year":"2005","journal-title":"J Gen Virol"},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.79.19.12495-12506.2005","article-title":"Envelope protein requirements for the assembly of infectious virions of porcine reproductive and respiratory syndrome virus","volume":"79","author":"Wissink","year":"2005","journal-title":"J Virol"},{"key":"B57","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.3434","article-title":"Gene-edited pigs are protected from porcine reproductive and respiratory syndrome virus","volume":"34","author":"Whitworth","year":"2016","journal-title":"Nat Biotechnol"},{"key":"B58","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00177-13","article-title":"An intact sialoadhesin (Sn\/SIGLEC1\/CD169) is not required for attachment\/internalization of the porcine reproductive and respiratory syndrome virus","volume":"87","author":"Prather","year":"2013","journal-title":"J Virol"},{"key":"B59","doi-asserted-by":"publisher","DOI":"10.1007\/s11262-008-0278-1","article-title":"Molecular assessment of the role of envelope-associated structural proteins in cross neutralization among different PRRS viruses","volume":"37","author":"Kim","year":"2008","journal-title":"Virus Genes"},{"key":"B60","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2016.02.016","article-title":"Efficacy of commercial genotype 1 porcine reproductive and respiratory syndrome virus (PRRSV) vaccine against field isolate of genotype 2 PRRSV","volume":"172","author":"Ko","year":"2016","journal-title":"Vet Immunol Immunopathol"},{"key":"B61","doi-asserted-by":"publisher","first-page":"e83567","DOI":"10.1371\/journal.pone.0083567","article-title":"Quantitative analysis of porcine reproductive and respiratory syndrome (PRRS) viremia profiles from experimental infection: a statistical modelling approach","volume":"8","author":"Islam","year":"2013","journal-title":"PloS One"},{"key":"B62","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1016\/j.virusres.2018.01.015","article-title":"Porcine reproductive and respiratory syndrome virus neutralizing antibodies provide in vivo cross-protection to PRRSV1 and PRRSV2 viral challenge","volume":"248","author":"Robinson","year":"2018","journal-title":"Virus Res"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2021.638493\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,2,22]],"date-time":"2021-02-22T05:03:39Z","timestamp":1613970219000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2021.638493\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,2,22]]},"references-count":62,"alternative-id":["10.3389\/fimmu.2021.638493"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2021.638493","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,2,22]]},"article-number":"638493"}}