{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,28]],"date-time":"2026-10-28T09:49:34Z","timestamp":1793180974811,"version":"4.5.0"},"reference-count":38,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,4,17]],"date-time":"2023-04-17T00:00:00Z","timestamp":1681689600000},"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. Microbiol."],"abstract":"<jats:p>Enterovirus A71 (EV-A71) can elicit a wide variety of human diseases such as hand, foot, and mouth disease and severe or fatal neurological complications. It is not clearly understood what determines the virulence and fitness of EV-A71. It has been observed that amino acid changes in the receptor binding protein, VP1, resulting in viral binding to heparan sulfate proteoglycans (HSPGs) may be important for the ability of EV-A71 to infect neuronal tissue. In this study, we identified that the presence of glutamine, as opposed to glutamic acid, at VP1-145 is key for viral infection in a 2D human fetal intestinal model, consistent with previous findings in an airway organoid model. Moreover, pre-treatment of EV-A71 particles with low molecular weight heparin to block HSPG-binding significantly reduced the infectivity of two clinical EV-A71 isolates and viral mutants carrying glutamine at VP1-145. Our data indicates that mutations in VP1 leading to HSPG-binding enhances viral replication in the human gut. These mutations resulting in increased production of viral particles at the primary replication site could lead to a higher risk of subsequent neuroinfection.<\/jats:p>\n                  <jats:sec>\n                    <jats:title>Importance<\/jats:title>\n                    <jats:p>With the near eradication of polio worldwide, polio-like illness (as is increasingly caused by EV-A71 infections) is of emerging concern. EV-A71 is indeed the most neurotropic enterovirus that poses a major threat globally to public health and specifically in infants and young children. Our findings will contribute to the understanding of the virulence and the pathogenicity of this virus. Further, our data also supports the identification of potential therapeutic targets against severe EV-A71 infection especially among infants and young children. Furthermore, our work highlights the key role of HSPG-binding mutations in the disease outcome of EV-A71. Additionally, EV-A71 is not able to infect the gut (the primary replication site in humans) in traditionally used animal models. Thus, our research highlights the need for human-based models to study human viral infections.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fmicb.2023.1045587","type":"journal-article","created":{"date-parts":[[2023,4,17]],"date-time":"2023-04-17T04:08:50Z","timestamp":1681704530000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":24,"title":["Amino acid variation at VP1-145 of enterovirus A71 determines the viral infectivity and receptor usage in a primary human intestinal model"],"prefix":"10.3389","volume":"14","author":[{"given":"Ikrame","family":"Aknouch","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"In\u00e9s","family":"Garc\u00eda-Rodr\u00edguez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesca Paola","family":"Giugliano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlemi","family":"Calitz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gerrit","family":"Koen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hetty","family":"van Eijk","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nina","family":"Johannessson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sjoerd","family":"Rebers","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lieke","family":"Brouwer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vanesa","family":"Muncan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Koert J.","family":"Stittelaar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dasja","family":"Pajkrt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katja C.","family":"Wolthers","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adithya","family":"Sridhar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,4,17]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"e202201432","DOI":"10.26508\/lsa.202201432","article-title":"Human milk inhibits some enveloped virus infections, including SARS-CoV-2, in an intestinal model","volume":"5","author":"Aknouch","year":"2022","journal-title":"Life Sci. Allian."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"2195","DOI":"10.1093\/nar\/18.8.2195","article-title":"Rapid purification of ribosomal RNAs from neutral agarose gels","volume":"18","author":"Boom","year":"1990","journal-title":"Nucleic Acids Res."},{"key":"ref3","doi-asserted-by":"publisher","first-page":"644","DOI":"10.1128\/JVI.74.2.644-651.2000","article-title":"Large-plaque mutants of sindbis virus show reduced binding to heparan sulfate, heightened viremia, and slower clearance from the circulation","volume":"74","author":"Byrnes","year":"2000","journal-title":"J. Virol."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"5250","DOI":"10.1128\/AAC.00443-15","article-title":"The agmatine-containing poly(amidoamine) polymer AGMA1 binds cell surface heparan sulfates and prevents attachment of mucosal human papillomaviruses","volume":"59","author":"Cagno","year":"2015","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"137","DOI":"10.1016\/j.antiviral.2004.07.001","article-title":"Protective effect of a natural carrageenan on genital herpes simplex virus infection in mice","volume":"64","author":"Carlucci","year":"2004","journal-title":"Antivir. Res."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"931","DOI":"10.1002\/jmv.23287","article-title":"Genetic characterization of enterovirus 71 isolated from patients with severe disease by comparative analysis of complete genomes","volume":"84","author":"Chang","year":"2012","journal-title":"J. Med. Virol."},{"key":"ref7","doi-asserted-by":"publisher","first-page":"1622","DOI":"10.1099\/vir.0.83676-0","article-title":"The molecular basis of mouse adaptation by human enterovirus 71","volume":"89","author":"Chua","year":"2008","journal-title":"J. Gen. Virol."},{"key":"ref8","doi-asserted-by":"publisher","first-page":"5816","DOI":"10.1128\/JVI.03826-13","article-title":"Cell surface vimentin is an attachment receptor for enterovirus 71","volume":"88","author":"du","year":"2014","journal-title":"J. Virol."},{"key":"ref9","doi-asserted-by":"publisher","first-page":"e00682-18","DOI":"10.1128\/JVI.00682-18","article-title":"VP1 amino acid residue 145 of enterovirus 71 is a key residue for its receptor attachment and resistance to neutralizing antibody during Cynomolgus monkey infection","volume":"92","author":"Fujii","year":"2018","journal-title":"J. Virol."},{"key":"ref10","doi-asserted-by":"publisher","first-page":"1288","DOI":"10.3390\/v12111288","article-title":"Put some guts into it: intestinal organoid models to study viral infection","volume":"12","author":"Garc\u00eda-Rodr\u00edguez","year":"2020","journal-title":"Viruses"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"740662","DOI":"10.3389\/fcimb.2021.740662","article-title":"Parechovirus a infection of the intestinal epithelium: differences between genotypes A1 and A3","volume":"11","author":"Garc\u00eda-Rodr\u00edguez","year":"2021","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref12","doi-asserted-by":"publisher","first-page":"eaau4255","DOI":"10.1126\/sciadv.aau4255","article-title":"Type III interferon signaling restricts enterovirus 71 infection of goblet cells","volume":"5","author":"Good","year":"2019","journal-title":"Sci. Adv."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"132","DOI":"10.1016\/j.virol.2011.10.015","article-title":"Mutations in VP2 and VP1 capsid proteins increase infectivity and mouse lethality of enterovirus 71 by virus binding and RNA accumulation enhancement","volume":"422","author":"Huang","year":"2012","journal-title":"Virology"},{"key":"ref14","doi-asserted-by":"publisher","first-page":"e1005033","DOI":"10.1371\/journal.ppat.1005033","article-title":"The role of VP1 amino acid residue 145 of enterovirus 71 in viral fitness and pathogenesis in a Cynomolgus monkey model","volume":"11","author":"Kataoka","year":"2015","journal-title":"PLoS Pathog."},{"key":"ref15","doi-asserted-by":"publisher","first-page":"e26237","DOI":"10.1371\/journal.pone.0026237","article-title":"Molecular analysis of virulent determinants of enterovirus 71","volume":"6","author":"Li","year":"2011","journal-title":"PLoS One"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"S52","DOI":"10.1086\/338819","article-title":"The 1998 enterovirus 71 outbreak in Taiwan: pathogenesis and management","volume":"34","author":"Lin","year":"2002","journal-title":"Clin. Infect. Dis."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1160\/TH07-04-0310","article-title":"Heparan sulfate-protein interactions: a concept for drug design?","volume":"98","author":"Lindahl","year":"2007","journal-title":"Thromb. Haemost."},{"key":"ref18","doi-asserted-by":"publisher","first-page":"794","DOI":"10.1038\/nm.1961","article-title":"Human P-selectin glycoprotein ligand-1 is a functional receptor for enterovirus 71","volume":"15","author":"Nishimura","year":"2009","journal-title":"Nat. Med."},{"key":"ref19","doi-asserted-by":"publisher","first-page":"2698","DOI":"10.1128\/JCM.00542-06","article-title":"Sensitive, seminested PCR amplification of VP1 sequences for direct identification of all enterovirus serotypes from original clinical specimens","volume":"44","author":"Nix","year":"2006","journal-title":"J. Clin. Microbiol."},{"key":"ref20","doi-asserted-by":"publisher","first-page":"1097","DOI":"10.1016\/S1474-4422(10)70209-X","article-title":"Clinical features, diagnosis, and management of enterovirus 71","volume":"9","author":"Ooi","year":"2010","journal-title":"Lancet Neurol."},{"key":"ref21","doi-asserted-by":"publisher","first-page":"373","DOI":"10.1007\/s004180100271","article-title":"Immunohistochemical localization of heparan sulfate proteoglycan in human gastrointestinal tract","volume":"115","author":"Oshiro","year":"2001","journal-title":"Histochem. Cell Biol."},{"key":"ref22","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1093\/oxfordjournals.aje.a118408","article-title":"A simple method of estimating fifty per cent endpoints","volume":"27","author":"Reed","year":"1938","journal-title":"Am. J. Epidemiol."},{"key":"ref23","doi-asserted-by":"publisher","first-page":"10","DOI":"10.3389\/fcimb.2020.00272","article-title":"A human 2D primary organoid-derived epithelial monolayer model to study host-pathogen interaction in the small intestine","volume":"10","author":"Roodsant","year":"2020","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref24","doi-asserted-by":"publisher","first-page":"1764","DOI":"10.3201\/eid1611.100104","article-title":"Enterovirus 71 infection with central nervous system involvement, South Korea","volume":"16","author":"Ryu","year":"2010","journal-title":"Emerg. Infect. Dis."},{"key":"ref25","doi-asserted-by":"publisher","first-page":"262","DOI":"10.1038\/nature07935","article-title":"Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche","volume":"459","author":"Sato","year":"2009","journal-title":"Nature"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"778","DOI":"10.1016\/S1473-3099(10)70194-8","article-title":"Virology, epidemiology, pathogenesis, and control of enterovirus 71","volume":"10","author":"Solomon","year":"2010","journal-title":"Lancet Infect. Dis."},{"key":"ref27","doi-asserted-by":"publisher","first-page":"1341","DOI":"10.3390\/v12111341","article-title":"A perspective on organoids for virology research","volume":"12","author":"Sridhar","year":"2020","journal-title":"Viruses"},{"key":"ref28","doi-asserted-by":"publisher","first-page":"12","DOI":"10.1186\/1471-2180-12-162","article-title":"Cell surface sialylation affects binding of enterovirus 71 to rhabdomyosarcoma and neuroblastoma cells","volume":"12","author":"Su","year":"2012","journal-title":"BMC Microbiol."},{"key":"ref29","doi-asserted-by":"publisher","first-page":"4527","DOI":"10.1128\/JVI.03498-14","article-title":"Cell surface Nucleolin facilitates enterovirus 71 binding and infection","volume":"89","author":"Su","year":"2015","journal-title":"J. Virol."},{"key":"ref30","doi-asserted-by":"publisher","first-page":"611","DOI":"10.1128\/JVI.02226-12","article-title":"Enterovirus 71 uses cell surface Heparan sulfate glycosaminoglycan as an attachment receptor","volume":"87","author":"Tan","year":"2013","journal-title":"J. Virol."},{"key":"ref31","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1016\/j.virol.2016.11.009","article-title":"VP1 residues around the five-fold axis of enterovirus A71 mediate heparan sulfate interaction","volume":"501","author":"Tan","year":"2017","journal-title":"Virology"},{"key":"ref32","doi-asserted-by":"publisher","first-page":"e1007190","DOI":"10.1371\/journal.ppat.1007190","article-title":"A VP1 mutation acquired during an enterovirus 71 disseminated infection confers heparan sulfate binding ability and modulates ex vivo tropism","volume":"14","author":"Tseligka","year":"2018","journal-title":"PLoS Pathog."},{"key":"ref33","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41426-018-0077-2","article-title":"Enterovirus 71 infection of human airway organoids reveals VP1-145 as a viral infectivity determinant","volume":"7","author":"van der Sanden","year":"2018","journal-title":"Emerg. Microb. Infect."},{"key":"ref34","doi-asserted-by":"publisher","first-page":"1062","DOI":"10.3724\/SP.J.1005.2014.1062","article-title":"The genetic control of mouse coat color and its applications in genetics teaching","volume":"36","author":"Xing","year":"2014","journal-title":"Yi Chuan"},{"key":"ref35","doi-asserted-by":"publisher","first-page":"798","DOI":"10.1038\/nm.1992","article-title":"Scavenger receptor B2 is a cellular receptor for enterovirus 71","volume":"15","author":"Yamayoshi","year":"2009","journal-title":"Nat. Med."},{"key":"ref36","doi-asserted-by":"publisher","first-page":"2351","DOI":"10.1128\/JCM.00563-09","article-title":"Enterovirus 71 outbreak in the People\u2019s Republic of China in 2008","volume":"47","author":"Yang","year":"2009","journal-title":"J. Clin. Microbiol."},{"key":"ref37","doi-asserted-by":"publisher","first-page":"541","DOI":"10.1007\/s00216-010-4117-6","article-title":"Hyphenated techniques for the analysis of heparin and heparan sulfate","volume":"399","author":"Yang","year":"2011","journal-title":"Anal. Bioanal. Chem."},{"key":"ref38","doi-asserted-by":"publisher","first-page":"14","DOI":"10.1186\/1471-2334-14-243","article-title":"The variations of VP1 protein might be associated with nervous system symptoms caused by enterovirus 71 infection","volume":"14","author":"Zhang","year":"2014","journal-title":"BMC Infect. Dis."}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2023.1045587\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,4,17]],"date-time":"2023-04-17T04:08:53Z","timestamp":1681704533000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2023.1045587\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,4,17]]},"references-count":38,"alternative-id":["10.3389\/fmicb.2023.1045587"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2023.1045587","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,4,17]]},"article-number":"1045587"}}