{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T04:32:50Z","timestamp":1790137970820,"version":"4.0.1"},"reference-count":38,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2017,4,19]],"date-time":"2017-04-19T00:00:00Z","timestamp":1492560000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2017,4,19]],"date-time":"2017-04-19T00:00:00Z","timestamp":1492560000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    HIV-1 envelope proteins (Envs) play a critical role in HIV infection. In a correct trimeric conformation, Env mediates virus\u2013cell binding and fusion. Malfunctioning of this machinery renders virions incapable of infecting cells. Each HIV-1 virion carries 10\u201314 Envs, and therefore a defective Env may not necessarily render a HIV virion non-infectious, since other Env on the same virion may still be functional. Alternatively, it is possible that on a given virion either all the spikes are defective or all are functional. Here, we investigate Env conformations on\n                    <jats:italic>individual<\/jats:italic>\n                    virions using our new nanotechnology, \u201cflow virometry\u201d, and a panel of antibodies that discriminate between various Env conformations. We found that the majority of HIV-1 virions carry either only trimeric (\u201cfunctional\u201d) or only defective spikes. The relatively small subfraction of virions that carry both functional and nonfunctional Envs contributes little to HIV infection of human lymphoid tissue\n                    <jats:italic>ex vivo<\/jats:italic>\n                    . The observation that the majority of virions exclusively express either functional or nonfunctional forms of Env has important implications for understanding the role of neutralizing and non-neutralizing antibodies in the immune control of HIV infection as well as for the development of effective prophylactic strategies.\n                  <\/jats:p>","DOI":"10.1038\/s41598-017-00935-w","type":"journal-article","created":{"date-parts":[[2017,4,11]],"date-time":"2017-04-11T07:58:52Z","timestamp":1491897532000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":30,"title":["Flow virometry analysis of envelope glycoprotein conformations on individual HIV virions"],"prefix":"10.1038","volume":"7","author":[{"given":"Anush","family":"Arakelyan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wendy","family":"Fitzgerald","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deborah F.","family":"King","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul","family":"Rogers","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hannah M.","family":"Cheeseman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jean-Charles","family":"Grivel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robin J.","family":"Shattock","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leonid","family":"Margolis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2017,4,19]]},"reference":[{"key":"935_CR1","doi-asserted-by":"publisher","first-page":"e1003046","DOI":"10.1371\/journal.pcbi.1003046","volume":"9","author":"A Sethi","year":"2013","unstructured":"Sethi, A., Tian, J., Derdeyn, C. A., Korber, B. & Gnanakaran, S. A mechanistic understanding of allosteric immune escape pathways in the HIV-1 envelope glycoprotein. PLoS computational biology \n                           9, e1003046, doi:10.1371\/journal.pcbi.1003046 (2013).","journal-title":"PLoS computational biology"},{"key":"935_CR2","doi-asserted-by":"publisher","first-page":"7932","DOI":"10.1128\/JVI.00757-08","volume":"82","author":"EM Bunnik","year":"2008","unstructured":"Bunnik, E. M., Pisas, L., van Nuenen, A. C. & Schuitemaker, H. Autologous neutralizing humoral immunity and evolution of the viral envelope in the course of subtype B human immunodeficiency virus type 1 infection. Journal of virology \n                           82, 7932\u20137941, doi:10.1128\/jvi.00757-08 (2008).","journal-title":"Journal of virology"},{"key":"935_CR3","doi-asserted-by":"publisher","first-page":"1631","DOI":"10.1128\/JVI.01482-09","volume":"84","author":"NA Doria-Rose","year":"2010","unstructured":"Doria-Rose, N. A. et al. Breadth of human immunodeficiency virus-specific neutralizing activity in sera: clustering analysis and association with clinical variables. Journal of virology \n                           84, 1631\u20131636, doi:10.1128\/jvi.01482-09 (2010).","journal-title":"Journal of virology"},{"key":"935_CR4","doi-asserted-by":"publisher","first-page":"866","DOI":"10.1038\/nm.1949","volume":"15","author":"L Stamatatos","year":"2009","unstructured":"Stamatatos, L., Morris, L., Burton, D. R. & Mascola, J. R. Neutralizing antibodies generated during natural HIV-1 infection: good news for an HIV-1 vaccine? Nature medicine \n                           15, 866\u2013870, doi:10.1038\/nm.1949 (2009).","journal-title":"Nature medicine"},{"key":"935_CR5","doi-asserted-by":"publisher","first-page":"7337","DOI":"10.1128\/JVI.00110-09","volume":"83","author":"MD Simek","year":"2009","unstructured":"Simek, M. D. et al. Human immunodeficiency virus type 1 elite neutralizers: individuals with broad and potent neutralizing activity identified by using a high-throughput neutralization assay together with an analytical selection algorithm. Journal of virology \n                           83, 7337\u20137348, doi:10.1128\/jvi.00110-09 (2009).","journal-title":"Journal of virology"},{"key":"935_CR6","doi-asserted-by":"publisher","first-page":"1884","DOI":"10.1126\/science.280.5371.1884","volume":"280","author":"R Wyatt","year":"1998","unstructured":"Wyatt, R. & Sodroski, J. The HIV-1 Envelope Glycoproteins: Fusogens, Antigens, and Immunogens. Science \n                           280, 1884\u20131888, doi:10.1126\/science.280.5371.1884 (1998).","journal-title":"Science"},{"key":"935_CR7","doi-asserted-by":"publisher","first-page":"1749","DOI":"10.1126\/science.8096089","volume":"259","author":"M Piatak Jr.","year":"1993","unstructured":"Piatak, M. Jr. et al. High levels of HIV-1 in plasma during all stages of infection determined by competitive PCR. Science \n                           259, 1749\u20131754, doi:10.1126\/science.8096089 (1993).","journal-title":"Science"},{"key":"935_CR8","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1016\/0092-8674(88)90487-4","volume":"53","author":"JM McCune","year":"1988","unstructured":"McCune, J. M. et al. Endoproteolytic cleavage of gp160 is required for the activation of human immunodeficiency virus. Cell \n                           53, 55\u201367, doi:10.1016\/0092-8674(88)90487-4 (1988).","journal-title":"Cell"},{"key":"935_CR9","doi-asserted-by":"publisher","first-page":"693","DOI":"10.1038\/nri3516","volume":"13","author":"PD Kwong","year":"2013","unstructured":"Kwong, P. D., Mascola, J. R. & Nabel, G. J. Broadly neutralizing antibodies and the search for an HIV-1 vaccine: the end of the beginning. Nat Rev Immunol \n                           13, 693\u2013701, doi:10.1038\/nri3516 (2013).","journal-title":"Nat Rev Immunol"},{"key":"935_CR10","doi-asserted-by":"publisher","first-page":"2515","DOI":"10.1128\/JVI.80.5.2515-2528.2006","volume":"80","author":"PL Moore","year":"2006","unstructured":"Moore, P. L. et al. Nature of nonfunctional envelope proteins on the surface of human immunodeficiency virus type 1. Journal of virology \n                           80, 2515\u20132528, doi:10.1128\/jvi.80.5.2515-2528.2006 (2006).","journal-title":"Journal of virology"},{"key":"935_CR11","doi-asserted-by":"crossref","unstructured":"Mao, Y. et al. Subunit organization of the membrane-bound HIV-1 envelope glycoprotein trimer. Nat Struct Mol Biol \n                           19, 893\u2013899, doi:http:\/\/www.nature.com\/nsmb\/journal\/v19\/n9\/abs\/nsmb.2351.html - supplementary-information (2012).","DOI":"10.1038\/nsmb.2351"},{"key":"935_CR12","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1186\/s12977-015-0207-z","volume":"12","author":"DJ Stieh","year":"2015","unstructured":"Stieh, D. J. et al. Discrete partitioning of HIV-1 Env forms revealed by viral capture. Retrovirology \n                           12, 81, doi:10.1186\/s12977-015-0207-z (2015).","journal-title":"Retrovirology"},{"key":"935_CR13","doi-asserted-by":"publisher","first-page":"416","DOI":"10.1038\/477416a","volume":"477","author":"PR Clapham","year":"2011","unstructured":"Clapham, P. R. & Lu, S. Vaccinology: Precisely tuned antibodies nab HIV. 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Journal of virology \n                           79, 1533\u20131542, doi:10.1128\/jvi.79.3.1533-1542.2005 (2005).","journal-title":"Journal of virology"},{"key":"935_CR16","doi-asserted-by":"publisher","first-page":"1707","DOI":"10.1093\/emboj\/cdg143","volume":"22","author":"JA Briggs","year":"2003","unstructured":"Briggs, J. A., Wilk, T., Welker, R., Krausslich, H. G. & Fuller, S. D. Structural organization of authentic, mature HIV-1 virions and cores. The EMBO journal \n                           22, 1707\u20131715, doi:10.1093\/emboj\/cdg143 (2003).","journal-title":"The EMBO journal"},{"key":"935_CR17","doi-asserted-by":"publisher","unstructured":"Lederle, A. et al. Neutralizing Antibodies Inhibit HIV-1 Infection of Plasmacytoid Dendritic Cells by an Fc\u03b3RIIa Independent Mechanism and Do Not Diminish Cytokines Production. Sci. Rep. 4, doi:10.1038\/srep05845 (2014).","DOI":"10.1038\/srep05845"},{"key":"935_CR18","doi-asserted-by":"crossref","first-page":"1100","DOI":"10.1128\/jvi.70.2.1100-1108.1996","volume":"70","author":"A Trkola","year":"1996","unstructured":"Trkola, A. et al. Human monoclonal antibody 2G12 defines a distinctive neutralization epitope on the gp120 glycoprotein of human immunodeficiency virus type 1. Journal of virology \n                           70, 1100\u20131108 (1996).","journal-title":"Journal of virology"},{"key":"935_CR19","doi-asserted-by":"publisher","first-page":"856","DOI":"10.1126\/science.1187659","volume":"329","author":"X Wu","year":"2010","unstructured":"Wu, X. et al. Rational design of envelope identifies broadly neutralizing human monoclonal antibodies to HIV-1. 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Science \n                           326, 285\u2013289, doi:10.1126\/science.1178746 (2009).","journal-title":"Science"},{"key":"935_CR22","doi-asserted-by":"publisher","first-page":"11231","DOI":"10.1128\/JVI.01543-12","volume":"86","author":"Y Li","year":"2012","unstructured":"Li, Y. et al. HIV-1 neutralizing antibodies display dual recognition of the primary and coreceptor binding sites and preferential binding to fully cleaved envelope glycoproteins. Journal of virology \n                           86, 11231\u201311241, doi:10.1128\/jvi.01543-12 (2012).","journal-title":"Journal of virology"},{"key":"935_CR23","doi-asserted-by":"publisher","first-page":"7306","DOI":"10.1128\/JVI.76.14.7306-7321.2002","volume":"76","author":"CN Scanlan","year":"2002","unstructured":"Scanlan, C. N. et al. The broadly neutralizing anti-human immunodeficiency virus type 1 antibody 2G12 recognizes a cluster of alpha1\u2013>2 mannose residues on the outer face of gp120. Journal of virology \n                           76, 7306\u20137321 (2002).","journal-title":"Journal of virology"},{"key":"935_CR24","doi-asserted-by":"publisher","first-page":"669","DOI":"10.1016\/j.immuni.2014.04.008","volume":"40","author":"C Blattner","year":"2014","unstructured":"Blattner, C. et al. Structural delineation of a quaternary, cleavage-dependent epitope at the gp41-gp120 interface on intact HIV-1 Env trimers. Immunity \n                           40, 669\u2013680, doi:10.1016\/j.immuni.2014.04.008 (2014).","journal-title":"Immunity"},{"key":"935_CR25","doi-asserted-by":"publisher","first-page":"811","DOI":"10.1126\/science.1192819","volume":"329","author":"T Zhou","year":"2010","unstructured":"Zhou, T. et al. Structural basis for broad and potent neutralization of HIV-1 by antibody VRC01. Science \n                           329, 811\u2013817, doi:10.1126\/science.1192819 (2010).","journal-title":"Science"},{"key":"935_CR26","doi-asserted-by":"publisher","first-page":"256","DOI":"10.1038\/nprot.2008.245","volume":"4","author":"J-C Grivel","year":"2009","unstructured":"Grivel, J.-C. & Margolis, L. Use of human tissue explants to study human infectious agents. Nat. Protocols \n                           4, 256\u2013269, doi:10.1038\/nprot.2008.245 (2009).","journal-title":"Nat. Protocols"},{"key":"935_CR27","doi-asserted-by":"publisher","first-page":"1183","DOI":"10.1111\/j.1462-5822.2012.01812.x","volume":"14","author":"PJ Klasse","year":"2012","unstructured":"Klasse, P. J. The molecular basis of HIV entry. Cellular microbiology \n                           14, 1183\u20131192, doi:10.1111\/j.1462-5822.2012.01812.x (2012).","journal-title":"Cellular microbiology"},{"key":"935_CR28","doi-asserted-by":"publisher","first-page":"407","DOI":"10.1084\/jem.174.2.407","volume":"174","author":"QJ Sattentau","year":"1991","unstructured":"Sattentau, Q. J. & Moore, J. P. Conformational changes induced in the human immunodeficiency virus envelope glycoprotein by soluble CD4 binding. The Journal of experimental medicine \n                           174, 407\u2013415, doi:10.1084\/jem.174.2.407 (1991).","journal-title":"The Journal of experimental medicine"},{"key":"935_CR29","doi-asserted-by":"publisher","first-page":"1151","DOI":"10.1038\/nbt745","volume":"20","author":"M Cavrois","year":"2002","unstructured":"Cavrois, M., De Noronha, C. & Greene, W. C. A sensitive and specific enzyme-based assay detecting HIV-1 virion fusion in primary T lymphocytes. Nature biotechnology \n                           20, 1151\u20131154, doi:10.1038\/nbt745 (2002).","journal-title":"Nature biotechnology"},{"key":"935_CR30","doi-asserted-by":"publisher","first-page":"12132","DOI":"10.1128\/JVI.79.19.12132-12147.2005","volume":"79","author":"X Yang","year":"2005","unstructured":"Yang, X., Kurteva, S., Ren, X., Lee, S. & Sodroski, J. Stoichiometry of envelope glycoprotein trimers in the entry of human immunodeficiency virus type 1. Journal of virology \n                           79, 12132\u201312147, doi:10.1128\/jvi.79.19.12132-12147.2005 (2005).","journal-title":"Journal of virology"},{"key":"935_CR31","doi-asserted-by":"publisher","first-page":"759","DOI":"10.1126\/science.1254426","volume":"346","author":"JB Munro","year":"2014","unstructured":"Munro, J. B. et al. Conformational dynamics of single HIV-1 envelope trimers on the surface of native virions. 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Virology \n                           488, 20\u201327, doi:10.1016\/j.virol.2015.10.021 (2016).","journal-title":"Virology"},{"key":"935_CR34","doi-asserted-by":"publisher","first-page":"489","DOI":"10.1016\/j.nano.2014.09.020","volume":"11","author":"A Arakelyan","year":"2015","unstructured":"Arakelyan, A., Ivanova, O., Vasilieva, E., Grivel, J. C. & Margolis, L. Antigenic composition of single nano-sized extracellular blood vesicles. Nanomedicine: nanotechnology, biology, and medicine \n                           11, 489\u2013498, doi:10.1016\/j.nano.2014.09.020 (2015).","journal-title":"Nanomedicine: nanotechnology, biology, and medicine"},{"key":"935_CR35","doi-asserted-by":"publisher","first-page":"439","DOI":"10.1084\/jem.20101907","volume":"208","author":"CR Ruprecht","year":"2011","unstructured":"Ruprecht, C. R. et al. MPER-specific antibodies induce gp120 shedding and irreversibly neutralize HIV-1. The Journal of experimental medicine \n                           208, 439\u2013454, doi:10.1084\/jem.20101907 (2011).","journal-title":"The Journal of experimental medicine"},{"key":"935_CR36","doi-asserted-by":"publisher","first-page":"e1005110","DOI":"10.1371\/journal.ppat.1005110","volume":"11","author":"LE McCoy","year":"2015","unstructured":"McCoy, L. E. et al. Incomplete Neutralization and Deviation from Sigmoidal Neutralization Curves for HIV Broadly Neutralizing Monoclonal Antibodies. PLoS pathogens \n                           11, e1005110, doi:10.1371\/journal.ppat.1005110 (2015).","journal-title":"PLoS pathogens"},{"key":"935_CR37","doi-asserted-by":"publisher","first-page":"5577","DOI":"10.1128\/JVI.74.12.5577-5586.2000","volume":"74","author":"P Greenhead","year":"2000","unstructured":"Greenhead, P. et al. Parameters of human immunodeficiency virus infection of human cervical tissue and inhibition by vaginal virucides. Journal of virology \n                           74, 5577\u20135586, doi:10.1128\/JVI.74.12.5577-5586.2000 (2000).","journal-title":"Journal of virology"},{"key":"935_CR38","doi-asserted-by":"publisher","first-page":"98","DOI":"10.1016\/j.jviromet.2008.11.013","volume":"157","author":"A Biancotto","year":"2009","unstructured":"Biancotto, A. et al. A highly sensitive and dynamic immunofluorescent cytometric bead assay for the detection of HIV-1 p24. Journal of virological methods \n                           157, 98\u2013101, doi:10.1016\/j.jviromet.2008.11.013 (2009).","journal-title":"Journal of virological methods"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-00935-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-00935-w","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-00935-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,22]],"date-time":"2022-12-22T22:43:35Z","timestamp":1671749015000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-00935-w"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,4,19]]},"references-count":38,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,12]]}},"alternative-id":["935"],"URL":"https:\/\/doi.org\/10.1038\/s41598-017-00935-w","relation":{"has-review":[{"id-type":"doi","id":"10.3410\/f.727535455.793531177","asserted-by":"object"}]},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,4,19]]},"assertion":[{"value":"3 October 2016","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"17 March 2017","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 April 2017","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare that they have no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"948"}}