{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T03:18:08Z","timestamp":1760239088336,"version":"build-2065373602"},"reference-count":38,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2020,10,8]],"date-time":"2020-10-08T00:00:00Z","timestamp":1602115200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP17fk0108306h0003"],"award-info":[{"award-number":["JP17fk0108306h0003"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001700","name":"Ministry of Education, Culture, Sports, Science and Technology","doi-asserted-by":"publisher","award":["17K10037"],"award-info":[{"award-number":["17K10037"]}],"id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Four kinds of avian-derived H5N1 influenza virus, A\/Vietnam\/1194\/2004 (Clade 1), A\/Indonesia\/5\/2005 (Clade 2.1), A\/Qinghai\/1A\/2005 (Clade 2.2), and A\/Anhui\/1\/2005 (Clade 2.3), have been stocked in Japan for use as pre-pandemic vaccines. When a pandemic occurs, these viruses would be used as vaccines in the hope of inducing immunity against the pandemic virus. We analyzed the specificity of antibodies (Abs) produced by B lymphocytes present in the blood after immunization with these vaccines. Eighteen volunteers took part in this project. After libraries of Ab-encoding sequences were constructed using blood from subjects vaccinated with these viruses, a large number of clones that encoded Abs that bound to the virus particles used as vaccines were isolated. These clones were classified into two groups according to the hemagglutination inhibition (HI) activity of the encoded Abs. While two-thirds of the clones were HI positive, the encoded Abs exhibited only restricted strain specificity. On the other hand, half of the HI-negative clones encoded Abs that bound not only to the H5N1 virus but also to the H1N1 virus; with a few exceptions, these Abs appeared to be encoded by memory B cells present before vaccination. The HI-negative clones included those encoding broadly cross-reactive Abs, some of which were encoded by non-VH1-69 germline genes. However, although this work shows that various kinds of anti-H5N1 Abs are encoded by volunteers vaccinated with pre-pandemic vaccines, broad cross-reactivity was seen only in a minority of clones, raising concern regarding the utility of these H5N1 vaccine viruses for the prevention of H5N1 pandemics.<\/jats:p>","DOI":"10.3390\/ijms21197422","type":"journal-article","created":{"date-parts":[[2020,10,8]],"date-time":"2020-10-08T09:39:55Z","timestamp":1602149995000},"page":"7422","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Comprehensive Analysis of Antibodies Induced by Vaccination with 4 Kinds of Avian Influenza H5N1 Pre-Pandemic Vaccines"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9076-1879","authenticated-orcid":false,"given":"Nobuko","family":"Ohshima","sequence":"first","affiliation":[{"name":"International Center for Cell and Gene Therapy, Research Promotion and Support Headquarters, Fujita Health University, Toyoake, Aichi 470-1192, Japan"},{"name":"Department of Innovation for Advanced Medicine, Center for Research Promotion and Support, Fujita Health University, Toyoake, Aichi 470-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoshitaka","family":"Iba","sequence":"additional","affiliation":[{"name":"International Center for Cell and Gene Therapy, Research Promotion and Support Headquarters, Fujita Health University, Toyoake, Aichi 470-1192, Japan"},{"name":"Department of Innovation for Advanced Medicine, Center for Research Promotion and Support, Fujita Health University, Toyoake, Aichi 470-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ritsuko","family":"Kubota-Koketsu","sequence":"additional","affiliation":[{"name":"Clinical Research Section, Research Department, The Research Foundation for Microbial Disease of Osaka University, Suita, Osaka 565-0871, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ayami","family":"Yamasaki","sequence":"additional","affiliation":[{"name":"Department of Innovation for Advanced Medicine, Center for Research Promotion and Support, Fujita Health University, Toyoake, Aichi 470-1192, Japan"},{"name":"Division of Innovation for Advanced Medicine, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keiko","family":"Majima","sequence":"additional","affiliation":[{"name":"Department of Innovation for Advanced Medicine, Center for Research Promotion and Support, Fujita Health University, Toyoake, Aichi 470-1192, Japan"},{"name":"Division of Innovation for Advanced Medicine, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9477-7467","authenticated-orcid":false,"given":"Gene","family":"Kurosawa","sequence":"additional","affiliation":[{"name":"International Center for Cell and Gene Therapy, Research Promotion and Support Headquarters, Fujita Health University, Toyoake, Aichi 470-1192, Japan"},{"name":"Department of Academic Research Support Promotion Facility, Center for Research Promotion and Support, Fujita Health University, Toyoake, Aichi 470-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daisuke","family":"Hirano","sequence":"additional","affiliation":[{"name":"Division of Rheumatology, Department of Internal Medicine, School of Medicine, Fujita Health University, Toyoake, Aichi 470-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shunji","family":"Yoshida","sequence":"additional","affiliation":[{"name":"Division of Rheumatology, Department of Internal Medicine, School of Medicine, Fujita Health University, Toyoake, Aichi 470-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mototaka","family":"Sugiura","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, School of Medicine, Fujita Health University, Toyoake, Aichi 470-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoshizo","family":"Asano","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, School of Medicine, Fujita Health University, Toyoake, Aichi 470-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoshinobu","family":"Okuno","sequence":"additional","affiliation":[{"name":"Osaka Institute of Public Health, Osaka 537-0025, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoshikazu","family":"Kurosawa","sequence":"additional","affiliation":[{"name":"Department of Innovation for Advanced Medicine, Center for Research Promotion and Support, Fujita Health University, Toyoake, Aichi 470-1192, Japan"},{"name":"Division of Innovation for Advanced Medicine, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,10,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1126\/science.279.5349.393","article-title":"Characterization of an avian influenza A (H5N1) virus isolated from a child with a fatal respiratory illness","volume":"279","author":"Subbarao","year":"1998","journal-title":"Science"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1016\/S0140-6736(97)11212-0","article-title":"Human influenza A H5N1 virus related to a highly pathogenic avian influenza virus","volume":"351","author":"Claas","year":"1998","journal-title":"Lancet"},{"doi-asserted-by":"crossref","unstructured":"Writing Committee of the World Health Organization (WHO) (2008). Consultation on Clinical Aspects of Human Infection with Avian Influenza A (H5N1) Virus. Update on avian influenza A (H5N1) virus infection in humans. N. Engl. J. Med., 358, 261\u2013273.","key":"ref_3","DOI":"10.1056\/NEJMra0707279"},{"unstructured":"World Health Organization Home Page (2020, August 25). Cumulative Number of Confirmed Human Cases of Avian Influenza A\/(H5N1) Reported to WHO. Available online: https:\/\/www.who.int\/influenza\/human_animal_interface\/H5N1_cumulative_table_archives\/en\/.","key":"ref_4"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1343","DOI":"10.1056\/NEJMoa055778","article-title":"Safety and immunogenicity of an inactivated subvirion influenza A (H5N1) vaccine","volume":"354","author":"Treanor","year":"2006","journal-title":"N. Engl. J. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1016\/S1473-3099(09)70106-9","article-title":"H5N1 influenza vaccination policy in Japan","volume":"9","author":"Masuda","year":"2009","journal-title":"Lancet Infect. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"4047","DOI":"10.1128\/JVI.03422-13","article-title":"Flow cytometry reveals that H5N1 vaccination elicits cross-reactive stem-directed antibodies from multiple Ig heavy-chain lineages","volume":"88","author":"Whittle","year":"2014","journal-title":"J. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"13260","DOI":"10.1128\/JVI.02133-14","article-title":"Induction of broadly reactive anti-hemagglutinin stalk antibodies by an H5N1 vaccine in humans","volume":"88","author":"Nachbagauer","year":"2014","journal-title":"J. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1093\/infdis\/jiw608","article-title":"Induction of broadly cross-reactive stalk-specific antibody responses to influenza group 1 and group 2 hemagglutinins by natural H7N9 virus infection in humans","volume":"215","author":"Liu","year":"2017","journal-title":"J. Infect. Dis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1026","DOI":"10.1016\/j.cell.2015.04.028","article-title":"Viral receptor-binding site antibodies with diverse germline origins","volume":"161","author":"Schmidt","year":"2015","journal-title":"Cell"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1255","DOI":"10.1172\/JCI74374","article-title":"Preexisting human antibodies neutralize recently emerged H7N9 influenza strains","volume":"125","author":"Dunand","year":"2015","journal-title":"J. Clin. Invest."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"S555","DOI":"10.1093\/infdis\/jix001","article-title":"Cross-reactive antibody responses to novel H5Nx influenza viruses following homologous and heterologous prime-boost vaccination with a prepandemic stockpiled A(H5N1) vaccine in humans","volume":"216","author":"Levine","year":"2017","journal-title":"J. Infect. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1016\/0022-2836(91)90498-U","article-title":"By-passing immunization. Human antibodies from V-gene libraries displayed on phage","volume":"222","author":"Marks","year":"1991","journal-title":"J. Mol. Biol."},{"key":"ref_14","first-page":"e502","article-title":"Vaccination with a recombinant H7 hemagglutinin-based influenza virus vaccine induces broadly reactive antibodies in humans","volume":"2","author":"Stadbauer","year":"2017","journal-title":"mSphere"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1016\/j.virol.2009.11.025","article-title":"Monoclonal antibodies in man that neutralized H3N2 influenza viruses were classified into three groups with distinct strain specificity: 1968\u20131973, 1977\u20131993 and 1997\u20132003","volume":"397","author":"Okada","year":"2010","journal-title":"Virology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1456","DOI":"10.1038\/nm.4224","article-title":"Molecular-level analysis of the serum antibody repertoire in young adults before and after seasonal influenza vaccination","volume":"22","author":"Lee","year":"2016","journal-title":"Nat. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1016\/S0022-1759(01)00462-8","article-title":"Antibody fusions with fluorescent proteins: A versatile reagent for profiling protein expression","volume":"257","author":"Morino","year":"2001","journal-title":"J. Immunol. Methods"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1016\/j.bbrc.2018.06.132","article-title":"Isolation of human monoclonal antibodies that bind to two different antigens and are encoded by germline VH and VL genes","volume":"503","author":"Ukai","year":"2018","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/S0378-1119(97)00101-7","article-title":"Expression vectors for the introduction of highly diverged sequences into the six complementarity-determining regions of an antibody","volume":"194","author":"Iba","year":"1997","journal-title":"Gene"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"11048","DOI":"10.1128\/JVI.05397-11","article-title":"Naturally occurring antibodies in humans can neutralize a variety of influenza virus strains, including H3, H1, H2 and H5","volume":"85","author":"Ohshima","year":"2011","journal-title":"J. Virol."},{"doi-asserted-by":"crossref","unstructured":"Ohshima, N., Kubota-Koketsu, R., Iba, Y., Okuno, Y., and Kurosawa, Y. (2014). Two types of antibodies are induced by vaccination with A\/California\/2009pdm virus: Binding near the sialic acid-binding pocket and neutralizing both H1N1 and H5N1 viruses. PLoS ONE, 9.","key":"ref_21","DOI":"10.1371\/journal.pone.0087305"},{"doi-asserted-by":"crossref","unstructured":"Hirano, D., Ohshima, N., Kubota-Koketsu, R., Yamasaki, A., Kurosawa, G., Okuno, Y., Yoshida, S., and Kurosawa, Y. (2018). Three types of broadly reacting antibodies against influenza B viruses induced by vaccination with seasonal influenza viruses. J. Immunol. Res.","key":"ref_22","DOI":"10.1155\/2018\/7251793"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1308","DOI":"10.1128\/jcm.28.6.1308-1313.1990","article-title":"Rapid focus reduction neutralization test of influenza A and B viruses in microtiter system","volume":"28","author":"Okuno","year":"1990","journal-title":"J. Clin. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"476","DOI":"10.1016\/j.antiviral.2013.03.021","article-title":"Broadly neutralizing antibodies against influenza viruses","volume":"98","author":"Laursen","year":"2013","journal-title":"Antivir. Res."},{"doi-asserted-by":"crossref","unstructured":"World Health Organization (WHO), World Organization for Animal Health (OIE), and Food and Agriculture Organization (FAO) (2014). Revised and updated nomenclature for highly pathogenic avian influenza A (H5N1) viruses. Influenza Other Respir. Viruses, 8, 384\u2013388.","key":"ref_25","DOI":"10.1111\/irv.12230"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1038\/333426a0","article-title":"Structure of the influenza virus haemagglutinin complexed with its receptor, sialic acid","volume":"333","author":"Weis","year":"1988","journal-title":"Nature"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2694","DOI":"10.1016\/j.jmb.2017.06.015","article-title":"A perspective on the structural and functional constraints for immune evasion: Insights from influenza virus","volume":"429","author":"Wu","year":"2017","journal-title":"J. Mol. Biol."},{"doi-asserted-by":"crossref","unstructured":"Burke, D.F., and Smith, D.J. (2014). A recommended numbering scheme for influenza A subtypes. PLoS ONE, 9.","key":"ref_28","DOI":"10.1371\/journal.pone.0112302"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1016\/0092-8674(82)90135-0","article-title":"The antigenic structure of the influenza virus A\/PR\/8\/34 hemagglutinin (H1 subtype)","volume":"31","author":"Caton","year":"1982","journal-title":"Cell"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1038\/290713a0","article-title":"Antigenic structure of influenza virus haemagglutinin defined by hybridoma antibodies","volume":"290","author":"Gerhard","year":"1981","journal-title":"Nature"},{"doi-asserted-by":"crossref","unstructured":"Throsby, M., van den Brink, E., Jongeneelen, M., Poon, L.L.M., Alard, P., Cornelissen, L., Bakker, A., Cox, F., van Deventer, E., and Guan, Y. (2008). Heterosubtypic neutralizing monoclonal antibodies cross-protective against H5N1 and H1N1 recovered from human IgM+ memory B cells. PLoS ONE, 3.","key":"ref_31","DOI":"10.1371\/journal.pone.0003942"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1126\/science.1171491","article-title":"Antibody recognition of a highly conserved influenza virus epitope","volume":"324","author":"Ekiert","year":"2009","journal-title":"Science"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1060","DOI":"10.1126\/science.1192517","article-title":"Induction of broadly neutralizing H1N1 influenza antibodies by vaccination","volume":"329","author":"Wei","year":"2010","journal-title":"Science"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1084\/jem.20101352","article-title":"Broadly cross-reactive antibodies dominate the human B cell response against 2009 pandemicH1N1 influenza virus infection","volume":"208","author":"Wrammert","year":"2011","journal-title":"J. Exp. Med."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1003","DOI":"10.1093\/cid\/cir121","article-title":"Wide prevalence of heterosubtypic broadly neutralizing human anti-influenza A antibodies","volume":"52","author":"Sui","year":"2011","journal-title":"Clin. Infect. Dis."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"850","DOI":"10.1126\/science.1205669","article-title":"A neutralizing antibody selected from plasma cells that binds to group 1 and group 2 influenza A hemagglutinins","volume":"333","author":"Corti","year":"2011","journal-title":"Science"},{"key":"ref_37","first-page":"323","article-title":"Structural characterization of viral epitopes recognized by broadly cross-reactive antibodies","volume":"386","author":"Lee","year":"2015","journal-title":"Curr. Top Microbiol. Immunol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1016\/j.cell.2016.06.043","article-title":"Vaccine-induced antibodies that neutralize group 1 and group 2 influenza A viruses","volume":"166","author":"Joyce","year":"2016","journal-title":"Cell"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/19\/7422\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:17:23Z","timestamp":1760177843000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/19\/7422"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,10,8]]},"references-count":38,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2020,10]]}},"alternative-id":["ijms21197422"],"URL":"https:\/\/doi.org\/10.3390\/ijms21197422","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2020,10,8]]}}}