{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T00:45:01Z","timestamp":1790729101625,"version":"4.1.0"},"reference-count":43,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2021,11,5]],"date-time":"2021-11-05T00:00:00Z","timestamp":1636070400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000774","name":"Defense Threat Reduction Agency","doi-asserted-by":"publisher","award":["HDTRA1-14-C-0115"],"award-info":[{"award-number":["HDTRA1-14-C-0115"]}],"id":[{"id":"10.13039\/100000774","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Defence Research and Development Canada-Medical Countermeasure Funds","award":["W1H-13-1-2-0"],"award-info":[{"award-number":["W1H-13-1-2-0"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The three encephalitic alphaviruses, namely, the Venezuelan, eastern, and western equine encephalitis viruses (VEEV, EEEV, and WEEV), are classified by the Centers for Disease Control and Prevention (CDC) as biothreat agents. Currently, no licensed medical countermeasures (MCMs) against these viruses are available for humans. Neutralizing antibodies (NAbs) are fast-acting and highly effective MCMs for use in both pre- and post-exposure settings against biothreat agents. While significant work has been done to identify anti-VEEV NAbs, less has been done to identify NAbs against EEEV and WEEV. In order to develop anti-EEEV or -WEEV NAbs, mice were immunized using complementary strategies with a variety of different EEEV or WEEV immunogens to maximize the generation of NAbs to each of these viruses. Of the hybridomas generated, three anti-EEEV and seven anti-WEEV monoclonal antibodies were identified with in vitro neutralization activity. The most potent neutralizers (two anti-EEEV NAbs and three anti-WEEV NAbs) were further evaluated for neutralization activity against additional strains of EEEV, a single strain of Madariaga virus (formerly South American EEEV), or WEEV. Of these, G1-2-H4 and G1-4-C3 neutralized all three EEEV strains and the Madariaga virus strain, whereas G8-2-H9 and 12 WA neutralized six out of eight WEEV strains. To determine the protective efficacy of these NAbs, the five most potent neutralizers were evaluated in respective mouse aerosol challenge models. All five NAbs demonstrated various levels of protection when administered at doses of 2.5 mg\/kg or 10 mg\/kg 24 h before the respective virus exposure via the aerosol route. Of these, anti-EEEV NAb G1-4-C3 and anti-WEEV NAb 8C2 provided 100% protection at both doses and all surviving mice were free of clinical signs throughout the study. Additionally, no virus was detected in the brain 14 days post virus exposure. Taken together, efficacious NAbs were developed that demonstrate the potential for the development of cross-strain antibody-based MCMs against EEEV and WEEV infections.<\/jats:p>","DOI":"10.3390\/v13112231","type":"journal-article","created":{"date-parts":[[2021,11,7]],"date-time":"2021-11-07T20:42:54Z","timestamp":1636317774000},"page":"2231","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Cross-Strain Neutralizing and Protective Monoclonal Antibodies against EEEV or WEEV"],"prefix":"10.3390","volume":"13","author":[{"given":"Amanda L.","family":"Phelps","sequence":"first","affiliation":[{"name":"Defence Science and Technology Laboratory, Porton Down, Salisbury SP4 0JQ, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lyn M.","family":"O\u2019Brien","sequence":"additional","affiliation":[{"name":"Defence Science and Technology Laboratory, Porton Down, Salisbury SP4 0JQ, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David O.","family":"Ulaeto","sequence":"additional","affiliation":[{"name":"Defence Science and Technology Laboratory, Porton Down, Salisbury SP4 0JQ, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frederick W.","family":"Holtsberg","sequence":"additional","affiliation":[{"name":"Integrated BioTherapeutics, Inc., Rockville, MD 20850, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grant C.","family":"Liao","sequence":"additional","affiliation":[{"name":"Integrated BioTherapeutics, Inc., Rockville, MD 20850, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robin","family":"Douglas","sequence":"additional","affiliation":[{"name":"Integrated BioTherapeutics, Inc., Rockville, MD 20850, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M. Javad","family":"Aman","sequence":"additional","affiliation":[{"name":"Integrated BioTherapeutics, Inc., Rockville, MD 20850, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pamela J.","family":"Glass","sequence":"additional","affiliation":[{"name":"US Army Medical Research Institute of Infectious Diseases, Frederick, MD 21702, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0398-6392","authenticated-orcid":false,"given":"Crystal L.","family":"Moyer","sequence":"additional","affiliation":[{"name":"Mapp Biopharmaceutical, Inc., San Diego, CA 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jane","family":"Ennis","sequence":"additional","affiliation":[{"name":"Mapp Biopharmaceutical, Inc., San Diego, CA 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6857-1024","authenticated-orcid":false,"given":"Larry","family":"Zeitlin","sequence":"additional","affiliation":[{"name":"Mapp Biopharmaceutical, Inc., San Diego, CA 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Les P.","family":"Nagata","sequence":"additional","affiliation":[{"name":"Defence Research and Development Canada, Suffield Research Centre, Medicine Hat, AB T1A 8K6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei-Gang","family":"Hu","sequence":"additional","affiliation":[{"name":"Defence Research and Development Canada, Suffield Research Centre, Medicine Hat, AB T1A 8K6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1016\/j.vetmic.2009.08.023","article-title":"Encephalitic alphaviruses","volume":"140","author":"Zacks","year":"2010","journal-title":"Vet. Microbiol."},{"key":"ref_2","unstructured":"Fields, B.N., Knipe, D.M., and Howley, P.M. (1996). Alphavirus. Fields Virology, Raven Publishers. [3rd ed.]."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1128\/mr.58.3.491-562.1994","article-title":"The alphaviruses: Gene expression, replication, and evolution","volume":"58","author":"Strauss","year":"1994","journal-title":"Microbiol. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3854","DOI":"10.1038\/emboj.2011.261","article-title":"4.4 A cryo-EM structure of an enveloped alphavirus Venezuelan equine encephalitis virus","volume":"30","author":"Zhang","year":"2011","journal-title":"EMBO J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1099\/0022-1317-44-3-725","article-title":"Biochemical and antigenic comparison of the envelope glycoproteins of Venezuelan equine encephalomyelitis virus strains","volume":"44","author":"France","year":"1979","journal-title":"J. Gen. Virol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1016\/0042-6822(82)90346-4","article-title":"Antigenic analysis of the surface glycoproteins of a Venezuelan equine encephalomyelitis virus (TC-83) using monoclonal antibodies","volume":"118","author":"Roehrig","year":"1982","journal-title":"Virology"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1014","DOI":"10.1128\/JVI.01586-09","article-title":"Evolutionary patterns of eastern equine encephalitis virus in North versus South America suggest ecological differences and taxonomic revision","volume":"84","author":"Arrigo","year":"2010","journal-title":"J. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"10682","DOI":"10.1128\/JVI.05032-11","article-title":"The role of the blood-brain barrier during Venezuelan equine encephalitis virus infection","volume":"85","author":"Schafer","year":"2011","journal-title":"J. Virol."},{"key":"ref_9","unstructured":"(2020, January 20). Breaking Down the 2019 Eastern Equine Encephalitis Outbreak in the US. Available online: https:\/\/www.contagionlive.com\/news\/breaking-down-the-2019-eastern-equine-encephalitis-outbreak-in-the-us."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"790","DOI":"10.1177\/0300985810372508","article-title":"Review paper: Pathology of animal models of alphavirus encephalitis","volume":"47","author":"Steele","year":"2010","journal-title":"Vet. Pathol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1283","DOI":"10.1126\/science.158.3806.1283","article-title":"Arbovirus infections of laboratory workers. Extent of problem emphasizes the need for more effective measures to reduce hazards","volume":"158","author":"Hanson","year":"1967","journal-title":"Science"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1016\/S0733-8627(01)00003-7","article-title":"Threats in bioterrorism. II: CDC category B and C agents","volume":"20","author":"Moran","year":"2002","journal-title":"Emerg. Med. Clin. N. Am."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1016\/S0166-3542(02)00203-6","article-title":"Viruses of the Bunya-and Togaviridae families: Potential as bioterrorism agents and means of control","volume":"57","author":"Sidwell","year":"2003","journal-title":"Antivir. Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"661","DOI":"10.2217\/fvl.13.42","article-title":"Vaccines and therapeutics for the encephalitic alphaviruses","volume":"8","author":"Nagata","year":"2013","journal-title":"Future Virol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"200","DOI":"10.3201\/eid0203.960306","article-title":"Antibody-based therapies for emerging infectious diseases","volume":"2","author":"Casadevall","year":"1996","journal-title":"Emerg. Infect. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1093\/clinids\/21.1.150","article-title":"Return to the past: The case for antibody-based therapies in infectious diseases","volume":"21","author":"Casadevall","year":"1995","journal-title":"Clin. Infect. Dis."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1016\/S1286-4579(00)00355-5","article-title":"Preventing infectious disease with passive immunization","volume":"2","author":"Zeitlin","year":"2000","journal-title":"Microbes Infect."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"54","DOI":"10.3201\/eid0501.990107","article-title":"Using monoclonal antibodies to prevent mucosal transmission of epidemic infectious diseases","volume":"5","author":"Zeitlin","year":"1999","journal-title":"Emerg. Infect. Dis."},{"key":"ref_19","unstructured":"Pollard, E. (2012, August 14). Mother, Daughter Taking Experimental Hendra virus Drugs. ABC News, Available online: http:\/\/www.abc.net.au\/news\/2010-05-27\/mother-daughter-taking-experimental-hendravirus\/844358."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"The PREVAIL II Writing Group, for the Multi-National PREVAIL II Study Team, Davey, R.T., Dodd, L., Proschan, M.A., Neaton, J., Neuhaus Nordwall, J., Koopmeiners, J.S., Beigel, J., and Tierney, J. (2016). A Randomized, Controlled Trial of ZMapp for Ebola Virus Infection. N. Engl. J. Med., 375, 1448\u20131456.","DOI":"10.1056\/NEJMoa1604330"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1320","DOI":"10.3389\/fmicb.2015.01320","article-title":"Approval of Raxibacumab for the Treatment of Inhalation Anthrax under the US Food and Drug Administration \u201cAnimal Rule\u201d","volume":"6","author":"Tsai","year":"2015","journal-title":"Front. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"IMpact-RSV Study Group (1998). Palivizumab, a humanized respiratory syncytial virus monoclonal antibody, reduces hospitalization from respiratory syncytial virus infection in high-risk infants. Pediatrics, 102, 531\u2013537.","DOI":"10.1542\/peds.102.3.531"},{"key":"ref_23","first-page":"1000149","article-title":"Opportunities and Challenges of Therapeutic Monoclonal Antibodies as Medical Countermeasures for Biodefense","volume":"7","author":"Hu","year":"2016","journal-title":"J. Bioterrorism Biodefense"},{"key":"ref_24","unstructured":"Hu, W.G., Nagata, L.P., and Vallerand, A. Novel Technology Platform of Therapeutic Antibody Development against Biothreat Agents: Cutting Edge Innovation for National Defence and Public Security. DRDC Scientific Letter 2015, DRDC-RDDC-2015-L189."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3210","DOI":"10.1016\/j.vaccine.2007.01.034","article-title":"Humanization and mammalian expression of a murine monoclonal antibody against Venezuelan equine encephalitis virus","volume":"25","author":"Hu","year":"2007","journal-title":"Vaccine"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"5558","DOI":"10.1016\/j.vaccine.2010.06.038","article-title":"A recombinant humanized monoclonal antibody completely protects mice against lethal challenge with Venezuelan equine encephalitis virus","volume":"28","author":"Hu","year":"2010","journal-title":"Vaccine"},{"key":"ref_27","unstructured":"Hu, W.G., Ngata, L.P., and Vallerand, A. Development of Fast-Acting, Long-Lasting, and Cost-Effective Medical Countermeasure against Venezuelan Equine Encephalitis Virus. DRDC Scientific Letter 2015, DRDC-RDDC-2016-L013."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"718","DOI":"10.4161\/mabs.28170","article-title":"Human-like antibodies neutralizing Western equine encephalitis virus","volume":"6","author":"Hulseweh","year":"2014","journal-title":"mAbs"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Burke, C.W., Froude, J.W., Miethe, S., Hulseweh, B., Hust, M., and Glass, P.J. (2018). Human-Like Neutralizing Antibodies Protect Mice from Aerosol Exposure with Western Equine Encephalitis Virus. Viruses, 10.","DOI":"10.3390\/v10040147"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3136","DOI":"10.1016\/j.celrep.2018.11.067","article-title":"Cryo-EM Structures of Eastern Equine Encephalitis Virus Reveal Mechanisms of Virus Disassembly and Antibody Neutralization","volume":"25","author":"Hasan","year":"2018","journal-title":"Cell Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1038\/s41564-018-0286-4","article-title":"Protective antibodies against Eastern equine encephalitis virus bind to epitopes in domains A and B of the E2 glycoprotein","volume":"4","author":"Kim","year":"2019","journal-title":"Nat. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2353","DOI":"10.1099\/vir.0.81815-0","article-title":"Infectivity variation and genetic diversity among strains of Western equine encephalitis virus","volume":"87","author":"Ngata","year":"2006","journal-title":"J. Gen. Virol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2280","DOI":"10.1016\/j.vaccine.2005.01.032","article-title":"Efficacy of DNA vaccination against western equine encephalitis virus infection","volume":"23","author":"Ngata","year":"2005","journal-title":"Vaccine"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1128\/JVI.02171-15","article-title":"Pan-ebolavirus and Pan-filovirus Mouse Monoclonal Antibodies: Protection against Ebola and Sudan Viruses","volume":"90","author":"Holtsberg","year":"2016","journal-title":"J. Virol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1016\/j.vetmic.2007.10.022","article-title":"Cloning, expression and purification of envelope proteins E1 and E2 of western equine encephalitis virus and potential use of them as antigens in immunoassays","volume":"128","author":"Hu","year":"2008","journal-title":"Vet. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1089\/hyb.2008.0011","article-title":"A novel approach to development of monoclonal antibodies using native antigen for immunization and recombinant antigen for screening","volume":"27","author":"Hu","year":"2008","journal-title":"Hybridoma"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"163","DOI":"10.3390\/v9070163","article-title":"Susceptibility and Lethality of Western Equine Encephalitis Virus in Balb\/c Mice When Infected by the Aerosol Route","volume":"9","author":"Phelps","year":"2017","journal-title":"Viruses"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1152\/ajplegacy.1947.150.1.70","article-title":"Measurement of the respiratory volumes of laboratory animals","volume":"150","author":"Guyton","year":"1947","journal-title":"Am. J. Physiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2091","DOI":"10.1099\/0022-1317-83-9-2091","article-title":"Occupancy and mechanism in antibody-mediated neutralization of animal viruses","volume":"83","author":"Klasse","year":"2002","journal-title":"J. Gen. Virol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1128\/jvi.52.1.290-292.1984","article-title":"Monoclonal antibodies to a monkeypox virus polypeptide determinant","volume":"52","author":"Roumillat","year":"1984","journal-title":"J. Virol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1038\/468645a","article-title":"Structural biology: An alphavirus puzzle solved","volume":"468","author":"Kielian","year":"2010","journal-title":"Nature"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/j.antiviral.2004.06.008","article-title":"Evaluation of a Western Equine Encephalitis recombinant E1 protein for protective immunity and diagnostics","volume":"64","author":"Das","year":"2004","journal-title":"Antivir. Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/j.virusres.2004.08.008","article-title":"Budding of alphaviruses","volume":"106","author":"Garoff","year":"2004","journal-title":"Virus Res."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/11\/2231\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:26:30Z","timestamp":1760167590000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/11\/2231"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,5]]},"references-count":43,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2021,11]]}},"alternative-id":["v13112231"],"URL":"https:\/\/doi.org\/10.3390\/v13112231","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,5]]}}}