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Infection by alphaviruses including CHIKV and the closely related Semliki Forest virus (SFV) can induce the formation of filopodia-like intercellular long extensions (ILEs). ILEs emanate from an infected cell, stably attach to a neighboring cell, and mediate cell-to-cell viral transmission that is resistant to neutralizing antibodies. However, our mechanistic understanding of ILE formation is limited, and the potential contribution of ILEs to CHIKV virulence or human CHIKV infection is unknown. Here, we used well-characterized virus mutants and monoclonal antibodies with known epitopes to dissect the virus requirements for ILE formation. Our results showed that both the viral E2 and E1 envelope proteins were required for ILE formation, while viral proteins 6K and transframe, and cytoplasmic nucleocapsid formation were dispensable. A subset of CHIKV monoclonal antibodies reduced ILE formation by masking specific regions particularly on the E2 A domain. Studies of the viral proteins from different CHIKV strains showed that ILE formation is conserved across the four major CHIKV lineages. Sera from convalescent human CHIKV patients inhibited ILE formation in cell culture, providing the first evidence for ILE inhibitory antibody production during human CHIKV infections.<\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>Chikungunya virus (CHIKV) infections can cause severe fever and long-lasting joint pain in humans. CHIKV is disseminated by mosquitoes and is now found world-wide, including in the Americas, Asia, and Africa. In cultured cells, CHIKV can induce the formation of long intercellular extensions that can transmit virus to another cell. However, our understanding of the formation of extensions and their importance in human CHIKV infection is limited. We here identified viral protein requirements for extension formation. We demonstrated that specific monoclonal antibodies against the virus envelope proteins or sera from human CHIKV patients can inhibit extension formation. Our data highlight the importance of evaluation of extension formation in the context of human CHIKV infection.<\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/mbio.01986-24","type":"journal-article","created":{"date-parts":[[2024,12,19]],"date-time":"2024-12-19T14:02:16Z","timestamp":1734616936000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":7,"title":["The alphavirus determinants of intercellular long extension formation"],"prefix":"10.1128","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9448-9182","authenticated-orcid":false,"given":"Caroline K.","family":"Martin","sequence":"first","affiliation":[{"name":"Department of Cell Biology, Albert Einstein College of Medicine","place":["Bronx, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Judy J.","family":"Wan","sequence":"additional","affiliation":[{"name":"Department of Cell Biology, Albert Einstein College of Medicine","place":["Bronx, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peiqi","family":"Yin","sequence":"additional","affiliation":[{"name":"Department of Cell Biology, Albert Einstein College of Medicine","place":["Bronx, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1811-2938","authenticated-orcid":true,"given":"Thomas E.","family":"Morrison","sequence":"additional","affiliation":[{"name":"Department of Immunology and Microbiology, University of Colorado School of Medicine","place":["Aurora, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William B.","family":"Messer","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, Oregon Health & Science University","place":["Portland, USA"]},{"name":"Department of Medicine, Division of Infectious Diseases, Oregon Health & Science University","place":["Portland, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vanessa","family":"Rivera-Amill","sequence":"additional","affiliation":[{"name":"Ponce Health Sciences University\/Ponce Research Institute","place":["Ponce, Puerto Rico"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4863-0015","authenticated-orcid":true,"given":"Jonathan R.","family":"Lai","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Albert Einstein College of Medicine","place":["Bronx, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nina","family":"Grau","sequence":"additional","affiliation":[{"name":"Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, CNRS UMR 3569, Unit\u00e9 de Virologie Structurale","place":["Paris, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9953-7988","authenticated-orcid":true,"given":"F\u00e9lix A.","family":"Rey","sequence":"additional","affiliation":[{"name":"Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, CNRS UMR 3569, Unit\u00e9 de Virologie Structurale","place":["Paris, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Th\u00e9r\u00e8se","family":"Couderc","sequence":"additional","affiliation":[{"name":"Institut Pasteur, Universite\u0301 Paris Cite\u0301, Inserm U1117, Biology of Infection Unit","place":["Paris, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc","family":"Lecuit","sequence":"additional","affiliation":[{"name":"Institut Pasteur, Universite\u0301 Paris Cite\u0301, Inserm U1117, Biology of Infection Unit","place":["Paris, France"]},{"name":"Department of Infectious Diseases and Tropical Medicine, Necker-Enfants Malades University Hospital, APHP, Institut Imagine","place":["Paris, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7395-4791","authenticated-orcid":true,"given":"Margaret","family":"Kielian","sequence":"additional","affiliation":[{"name":"Department of Cell Biology, Albert Einstein College of Medicine","place":["Bronx, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_5_2_2","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms8081167"},{"key":"e_1_3_5_3_2","first-page":"170","volume-title":"Fields virology","author":"Kuhn RJ","year":"2021","unstructured":"Kuhn RJ. 2021. 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