{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T01:40:37Z","timestamp":1768527637973,"version":"3.49.0"},"posted":{"date-parts":[[2025,10,3]]},"group-title":"Immunology","reference-count":32,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,10,3]],"date-time":"2025-10-03T00:00:00Z","timestamp":1759449600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,10,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>To evaluate the immunogenicity of lipooligosaccharide (LOS) based gonococcal vaccines, we developed a multiplexed fluorescent microsphere immunoassay that distinguishes and quantifies antibodies that bind the various LOS antigens.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>We coupled LOS from gonococcal strains 1291wt (nLc4 \u03b1 chain), 1291a (Lc3 \u03b1 chain) and 1291c (Lc2 \u03b1 chain), to different fluorescent microsphere regions (BioRad) after disaggregation in 1% deoxycholate. The microspheres were incubated with sera from 37 participants in a study of gonococcal risk factors. Laser excited fluorescence from each region was converted to ng\/mL of IgG with use of affinity purified human IgG specific for each LOS. IgG specific to the nLc4 terminal galactose was quantified by subtraction of Lc3 IgG from 1291wt IgG; that specific to the Lc3 terminal glucosamine by subtraction of Lc2 IgG from Lc3 IgG.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    Concentrations of nLc4, Lc3 and Lc2 IgG summed to the concentration of 1291wt IgG. Visual inspection of the data revealed a non-normally distributed, bimodal distribution of nLc4 IgG concentrations; 31\/37 had a mean serum nLc4 IgG concentration of 6.22 ng\/mL, and the other six had a mean nLc4 IgG concentration of 11.2 ng\/ml. Upon statistical comparison of these groups, conducted prior to knowledge of infection status, those participants whose nLc4 IgG concentration centered around a higher mean, were more likely to be asymptomatic (\n                    <jats:italic>p<\/jats:italic>\n                    = 0.03) and were less likely to be infected (\n                    <jats:italic>p<\/jats:italic>\n                    = 0.05) than those with lower IgG concentrations.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>This accurate LOS immunoassay can be expanded to include any number of LOS specificities.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Short Summary<\/jats:title>\n                  <jats:p>\n                    We developed a multiplexed fluorescent microsphere immunoassay of IgG antibodies to glycose antigens of the\n                    <jats:italic>Neisseria<\/jats:italic>\n                    Lipooligosaccharide in clients of an STI clinic who were sexually exposed to gonorrhea.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.10.01.679747","type":"posted-content","created":{"date-parts":[[2025,10,3]],"date-time":"2025-10-03T09:10:24Z","timestamp":1759482624000},"source":"Crossref","is-referenced-by-count":0,"title":["A Multiplexed Fluorescent Microsphere Immunoassay of IgG Antibodies to the Individual Glycose Antigens of the\n                  <i>Neisseria<\/i>\n                  Lipooligosaccharide \u03b1 Chain in Clients of an STI Clinic"],"prefix":"10.64898","author":[{"given":"Stephanie E.","family":"McLaughlin","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhijie","family":"Yang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J. McLeod","family":"Griffiss","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025100610250750000_2025.10.01.679747v1.1","doi-asserted-by":"publisher","DOI":"10.1097\/00007435-200110000-00001"},{"key":"2025100610250750000_2025.10.01.679747v1.2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.174.6.1601"},{"key":"2025100610250750000_2025.10.01.679747v1.3","doi-asserted-by":"crossref","unstructured":"Barrier BF , Gargiulo AR , Schust DJ. Reproductive Immunology and Its Disorders. Chapter 13 In: Yen & Jaffe\u2019s Reproductive Endocrinology. Strauss JF, Barbieri RL, editors. 6th ed. 2009. p. 299\u2013323.","DOI":"10.1016\/B978-1-4160-4907-4.00013-9"},{"key":"2025100610250750000_2025.10.01.679747v1.4","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/10.Supplement_2.S287"},{"issue":"38","key":"2025100610250750000_2025.10.01.679747v1.5","first-page":"461","article-title":"The Role of Bacterial Lipooligosaccharides in the Pathogenesis of Human Disease","volume":"7","year":"2010","journal-title":"Trends Glycosci Glycotechnol"},{"key":"2025100610250750000_2025.10.01.679747v1.6","unstructured":"Griffiss JMcL , Schneider H. Endotoxin in Health and Disease. The chemistry and biology of lipooli-gosaccharides: The endotoxins of bacteria of the respiratory and genital mucosae. New York: Marcel Dekker; p. 174\u2013194."},{"issue":"1","key":"2025100610250750000_2025.10.01.679747v1.7","first-page":"107","article-title":"Lipooligosaccharides (LOS) of Neisseria gonorrhoeae and Neisseria meningitidis have components that are immunochemically similar to precursors of human blood group antigens","volume":"168","year":"1988","journal-title":"Carbohydrate sequence specificity of the mouse monoclonal antibodies that recognize crossreacting antigens on LOS and human erythrocytes. J Exp Medicine"},{"key":"2025100610250750000_2025.10.01.679747v1.8","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jis481"},{"key":"2025100610250750000_2025.10.01.679747v1.9","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.291583"},{"key":"2025100610250750000_2025.10.01.679747v1.10","doi-asserted-by":"publisher","DOI":"10.1084\/jem.183.1.323"},{"issue":"8","key":"2025100610250750000_2025.10.01.679747v1.11","doi-asserted-by":"crossref","first-page":"1792","DOI":"10.1128\/iai.55.8.1792-1800.1987","article-title":"Physical heterogeneity of neisserial lipooligosaccharides reflects oligosaccharides that differ in apparent molecular weight, chemical composition, and antigenic expression","volume":"55","year":"1987","journal-title":"Infect Immun"},{"key":"2025100610250750000_2025.10.01.679747v1.12","doi-asserted-by":"publisher","DOI":"10.1016\/0161-5890(88)90116-2"},{"key":"2025100610250750000_2025.10.01.679747v1.13","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.05125-11"},{"key":"2025100610250750000_2025.10.01.679747v1.14","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01445-09"},{"issue":"29","key":"2025100610250750000_2025.10.01.679747v1.15","doi-asserted-by":"crossref","first-page":"19303","DOI":"10.1016\/S0021-9258(18)54998-X","article-title":"The structural basis for pyocin resistance in Neisseria gonorrhoeae lipooligosaccharides","volume":"266","year":"1991","journal-title":"J Biol Chem"},{"key":"2025100610250750000_2025.10.01.679747v1.16","doi-asserted-by":"publisher","DOI":"10.1097\/OLQ.0000000000000926"},{"issue":"3","key":"2025100610250750000_2025.10.01.679747v1.17","doi-asserted-by":"crossref","first-page":"842","DOI":"10.4049\/jimmunol.116.3.842","article-title":"Response to Antigenic Determinants of Neisseria Meningitidis Lipopolysaccharide Investigated with A New Radioactive Antigen-Binding Assay","volume":"116","year":"1976","journal-title":"J Immunol"},{"key":"2025100610250750000_2025.10.01.679747v1.18","unstructured":"Bio-Rad. Multiplex Immunoassays. [Internet]. [cited 2024 Aug 17]. Available from: www.bio-rad.com\/en-us\/applicationtechnologies\/multiplex-immunoassays?ID=LUSM0E8UU"},{"key":"2025100610250750000_2025.10.01.679747v1.19","doi-asserted-by":"publisher","DOI":"10.1016\/0002-9343(83)91112-9"},{"key":"2025100610250750000_2025.10.01.679747v1.20","doi-asserted-by":"publisher","DOI":"10.1309\/LMCH-C4Q2-VFL9-3T1A"},{"key":"2025100610250750000_2025.10.01.679747v1.21","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01280-08"},{"key":"2025100610250750000_2025.10.01.679747v1.22","unstructured":"Wang J. Neisseria gonorrhoeae must express the paraglobosyl LOS in order to invade human genitourinary epithelial cells. In: WD Z, CE F, CD D, editors. Abstracts of the Tenth International Pathogenic Neisseria Conference. Baltimore, Maryland; 1996."},{"key":"2025100610250750000_2025.10.01.679747v1.23","doi-asserted-by":"publisher","DOI":"10.1084\/jem.191.6.949"},{"key":"2025100610250750000_2025.10.01.679747v1.24","doi-asserted-by":"crossref","unstructured":"Gulati S , Shaughnessy J , Ram S , Rice PA . Targeting Lipooligosaccharide (LOS) for a Gonococcal Vaccine. Front Immunol. 2019; 10:321.","DOI":"10.3389\/fimmu.2019.00321"},{"key":"2025100610250750000_2025.10.01.679747v1.25","doi-asserted-by":"publisher","DOI":"10.3201\/eid2206.151093"},{"key":"2025100610250750000_2025.10.01.679747v1.26","doi-asserted-by":"publisher","DOI":"10.1097\/00007435-200007000-00005"},{"key":"2025100610250750000_2025.10.01.679747v1.27","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/174.Supplement_2.S134"},{"key":"2025100610250750000_2025.10.01.679747v1.28","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/155.6.1276"},{"key":"2025100610250750000_2025.10.01.679747v1.29","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.67.8.3937-3946.1999"},{"key":"2025100610250750000_2025.10.01.679747v1.30","doi-asserted-by":"publisher","DOI":"10.1081\/IAS-120003658"},{"issue":"12","key":"2025100610250750000_2025.10.01.679747v1.31","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.4161\/hv.6.12.13399","article-title":"Correlation between levels of human papillomavirus (HPV)-16 and 18 antibodies in serum and cervicovaginal secretions in girls and women vaccinated with the HPV-16\/18 AS04-adjuvanted vaccine","volume":"6","year":"2010","journal-title":"Hum Vaccines"},{"issue":"25","key":"2025100610250750000_2025.10.01.679747v1.32","doi-asserted-by":"crossref","first-page":"2389","DOI":"10.1056\/NEJMe2311948","article-title":"Doxycycline Postexposure Prophylaxis for STIs in Women \u2014 Uncertain Benefit, Urgent Need","volume":"389","year":"2023","journal-title":"N Engl J Med"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.10.01.679747","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T09:43:13Z","timestamp":1768470193000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.10.01.679747"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,3]]},"references-count":32,"URL":"https:\/\/doi.org\/10.1101\/2025.10.01.679747","relation":{},"subject":[],"published":{"date-parts":[[2025,10,3]]},"subtype":"preprint"}}