{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,17]],"date-time":"2026-03-17T16:19:58Z","timestamp":1773764398766,"version":"3.50.1"},"reference-count":48,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,3,17]],"date-time":"2026-03-17T00:00:00Z","timestamp":1773705600000},"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. Mol. Neurosci."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Patients with glycine receptor (GlyR) aAbs suffer from various diseases, including stiff-person syndrome (SPS), and currently, no cure exists. Several treatment options exist; however, these treatment options lack specificity. To date, only one common epitope has been mapped for GlyR aAbs in the far N-terminal region of the GlyR\u03b11 subunit. However, some patient sera also bind GlyR\u03b12, GlyR\u03b13, or GlyR\u03b2. Therefore, more than one common epitope may exist. Unraveling these epitopes will help generate more specific treatment approaches.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Here, we constructed GlyRa1 and GlyRa3 variants by site-directed mutagenesis using amino acid differences between these two subunits within their extracellular domains. Peptide microarrays, which have shown that an epitope including the binding site of a commercial pan-a antibody (\n                      <jats:sup>96<\/jats:sup>\n                      PDLFFANEKS\n                      <jats:sup>105<\/jats:sup>\n                      ) and its surrounding residues is highly relevant for aAb binding, were utilized to identify additional residues important for aAb binding. Two overlapping peptides (\n                      <jats:sup>93<\/jats:sup>\n                      LWKPDLFFANEKSAN\n                      <jats:sup>107<\/jats:sup>\n                      and\n                      <jats:sup>98<\/jats:sup>\n                      LFFANEKSANFHDVT\n                      <jats:sup>112<\/jats:sup>\n                      ) were used for aAb neutralization in cell-based assays.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The GlyRa1 and GlyRa3 variants helped to identify which amino acid sequences in the extracellular domain of GlyRs represent additional aAb epitopes or are involved in aAb binding. Using both generated peptides for aAb neutralization with a patient serum containing GlyRb aAbs that bind specifically to this region\n                      <jats:sup>96<\/jats:sup>\n                      PDLFFANEKSANFHDV\n                      <jats:sup>111<\/jats:sup>\n                      , successful neutralization was demonstrated. In contrast, when using patient sera that reliably target the extracellular domain including\n                      <jats:sup>96<\/jats:sup>\n                      PDLFFANEKS\n                      <jats:sup>105<\/jats:sup>\n                      of the GlyRa subunits, the overlapping peptides reduced aAb binding but failed to fully neutralize the aAbs.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>In conclusion, our data demonstrate that GlyR aAbs are polyclonal or bind to structural epitopes. These results define single residues important for aAb binding and help explain why no further common aAb binding site has been identified so far. Hence, patient-specific pattern for GlyR aAbs exist, emphasizing the importance of epitope characterization as basis for future therapeutic testing or even complete neutralization of the aAbs.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fnmol.2026.1747209","type":"journal-article","created":{"date-parts":[[2026,3,17]],"date-time":"2026-03-17T14:20:17Z","timestamp":1773757217000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Polyclonal glycine receptor aAbs: a challenge for personalized epitope characterization"],"prefix":"10.3389","volume":"19","author":[{"given":"Anna-Lena","family":"Wiessler","sequence":"first","affiliation":[{"name":"Institute for Clinical Neurobiology, University Hospital W\u00fcrzburg","place":["W\u00fcrzburg, Germany"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Inken","family":"Stahl","sequence":"additional","affiliation":[{"name":"Department of Neurology, University Hospital W\u00fcrzburg","place":["W\u00fcrzburg, Germany"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Natascha","family":"Schaefer","sequence":"additional","affiliation":[{"name":"Institute for Clinical Neurobiology, University Hospital W\u00fcrzburg","place":["W\u00fcrzburg, Germany"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vera","family":"Rauschenberger","sequence":"additional","affiliation":[{"name":"Institute for Clinical Neurobiology, University Hospital W\u00fcrzburg","place":["W\u00fcrzburg, Germany"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ivan","family":"Talucci","sequence":"additional","affiliation":[{"name":"Rudolf Virchow Center for Integrative and Translational Bioimaging, University of W\u00fcrzburg","place":["W\u00fcrzburg, Germany"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hans M.","family":"Maric","sequence":"additional","affiliation":[{"name":"Rudolf Virchow Center for Integrative and Translational Bioimaging, University of W\u00fcrzburg","place":["W\u00fcrzburg, Germany"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bet\u00fcl","family":"Baykan","sequence":"additional","affiliation":[{"name":"Department of Neurology and Clinical Neurophysiology, EMAR Medical Center","place":["Istanbul, T\u00fcrkiye"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Erdem","family":"T\u00fcz\u00fcn","sequence":"additional","affiliation":[{"name":"Institute of Experimental Medicine, Istanbul University","place":["Istanbul, T\u00fcrkiye"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claudia","family":"Sommer","sequence":"additional","affiliation":[{"name":"Department of Neurology, University Hospital W\u00fcrzburg","place":["W\u00fcrzburg, Germany"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carmen","family":"Villmann","sequence":"additional","affiliation":[{"name":"Institute for Clinical Neurobiology, University Hospital W\u00fcrzburg","place":["W\u00fcrzburg, Germany"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2026,3,17]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"2178","DOI":"10.1093\/brain\/awu142","article-title":"Glycine receptor antibodies in PERM and related syndromes: characteristics, clinical features and outcomes","volume":"137","author":"Carvajal-Gonzalez","year":"2014","journal-title":"Brain"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"3398","DOI":"10.1093\/brain\/awz297","article-title":"Glycine receptor autoantibodies disrupt inhibitory neurotransmission","volume":"142","author":"Crisp","year":"2019","journal-title":"Brain"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"48","DOI":"10.1007\/s11910-008-0009-y","article-title":"Advances in the pathogenesis and treatment of patients with stiff person syndrome","volume":"8","author":"Dalakas","year":"2008","journal-title":"Curr. Neurol. Neurosci. Rep."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"W13","DOI":"10.1093\/nar\/gkr245","article-title":"T-coffee: a web server for the multiple sequence alignment of protein and RNA sequences using structural information and homology extension","volume":"39","author":"Di Tommaso","year":"2011","journal-title":"Nucleic Acids Res."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"e186","DOI":"10.1212\/nxi.0000000000000186","article-title":"Lockjaw in stiff-person syndrome with autoantibodies against glycine receptors","volume":"3","author":"Doppler","year":"2016","journal-title":"Neurology Neuroimmunol. Neuroinflammation"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"40216","DOI":"10.1074\/jbc.R112.408229","article-title":"Inhibitory glycine receptors: an update","volume":"287","author":"Dutertre","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref7","doi-asserted-by":"publisher","first-page":"414","DOI":"10.1111\/epi.12528","article-title":"Investigation of neuronal autoantibodies in two different focal epilepsy syndromes","volume":"55","author":"Ekizoglu","year":"2014","journal-title":"Epilepsia"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"139","DOI":"10.4137\/CCRep.S16941","article-title":"Stiff man syndrome: a diagnostic dilemma in a young female with diabetes mellitus and thyroiditis","volume":"7","author":"Enuh","year":"2014","journal-title":"Clin Med Insights Case Rep"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"e200507","DOI":"10.1212\/NXI.0000000000200507","article-title":"Distinct epitopes are associated with clinical phenotypes in autoimmune nodopathies with anti-Contactin1 autoantibodies","volume":"13","author":"Gr\u00fcner","year":"2025","journal-title":"Neurol. Neuroimmunol. Neuroinflamm."},{"key":"ref10","doi-asserted-by":"publisher","first-page":"18207","DOI":"10.1073\/pnas.0509024102","article-title":"Molecular tuning of fast gating in pentameric ligand-gated ion channels","volume":"102","author":"Grutter","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"472","DOI":"10.1080\/08916934.2016.1214823","article-title":"Effect of therapeutic plasma exchange on immunoglobulins in myasthenia gravis","volume":"49","author":"Guptill","year":"2016","journal-title":"Autoimmunity"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"4479","DOI":"10.4049\/jimmunol.173.7.4479","article-title":"CpG DNA induces IgG class switch DNA recombination by activating human B cells through an innate pathway that requires TLR9 and cooperates with IL-10","volume":"173","author":"He","year":"2004","journal-title":"J. Immunol."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"e438","DOI":"10.1212\/NXI.0000000000000438","article-title":"Glycine receptor modulating antibody predicting treatable stiff-person spectrum disorders","volume":"5","author":"Hinson","year":"2018","journal-title":"Neurol. Neuroimmunol. Neuroinflamm."},{"key":"ref14","doi-asserted-by":"crossref","first-page":"1089","DOI":"10.1002\/eji.1830180718","article-title":"Autoantibody production is associated with polyclonal B cell activation in autoimmune mice which express the lpr or gld genes","volume":"18","author":"Ishigatsubo","year":"1988","journal-title":"Eur. J. Immunol."},{"key":"ref15","doi-asserted-by":"publisher","first-page":"2131","DOI":"10.1039\/c0cs00029a","article-title":"Studying protein-protein interactions using peptide arrays","volume":"40","author":"Katz","year":"2011","journal-title":"Chem. Soc. Rev."},{"key":"ref16","doi-asserted-by":"publisher","first-page":"405","DOI":"10.1002\/ana.25666","article-title":"Human cerebrospinal fluid monoclonal LGI1 autoantibodies increase neuronal excitability","volume":"87","author":"Kornau","year":"2020","journal-title":"Ann. Neurol."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"2641","DOI":"10.1093\/brain\/aww208","article-title":"Human cerebrospinal fluid monoclonal N-methyl-D-aspartate receptor autoantibodies are sufficient for encephalitis pathogenesis","volume":"139","author":"Kreye","year":"2016","journal-title":"Brain"},{"key":"ref18","doi-asserted-by":"publisher","first-page":"e20210012","DOI":"10.1084\/jem.20210012","article-title":"Encephalitis patient-derived monoclonal GABAA receptor antibodies cause epileptic seizures","volume":"218","author":"Kreye","year":"2021","journal-title":"J. Exp. Med."},{"key":"ref19","doi-asserted-by":"publisher","first-page":"e2320","DOI":"10.1016\/j.cell.2025.03.004","article-title":"Autoimmune mechanisms elucidated through muscle acetylcholine receptor structures","volume":"188","author":"Li","year":"2025","journal-title":"Cell"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"1051","DOI":"10.1152\/physrev.00042.2003","article-title":"Molecular structure and function of the glycine receptor chloride channel","volume":"84","author":"Lynch","year":"2004","journal-title":"Physiol. Rev."},{"key":"ref21","doi-asserted-by":"publisher","first-page":"2401","DOI":"10.1002\/j.1460-2075.1991.tb07779.x","article-title":"Widespread expression of glycine receptor subunit mRNAs in the adult and developing rat brain","volume":"10","author":"Malosio","year":"1991","journal-title":"EMBO J."},{"key":"ref22","doi-asserted-by":"publisher","first-page":"714","DOI":"10.1001\/jamaneurol.2016.0133","article-title":"Clinical and immunologic investigations in patients with stiff-person Spectrum disorder","volume":"73","author":"Martinez-Hernandez","year":"2016","journal-title":"JAMA Neurol."},{"key":"ref23","doi-asserted-by":"crossref","first-page":"19708","DOI":"10.1074\/jbc.273.31.19708","article-title":"The human glycine receptor subunit alpha3. Glra3 gene structure, chromosomal localization, and functional characterization of alternative transcripts","volume":"273","author":"Nikolic","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref24","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1006\/jmbi.2000.4042","article-title":"T-coffee: a novel method for fast and accurate multiple sequence alignment","volume":"302","author":"Notredame","year":"2000","journal-title":"J. Mol. Biol."},{"key":"ref25","doi-asserted-by":"publisher","first-page":"2469","DOI":"10.1016\/j.cell.2022.06.025","article-title":"Structural mechanisms of GABA(a) receptor autoimmune encephalitis","volume":"185","author":"Noviello","year":"2022","journal-title":"Cell"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"1851","DOI":"10.1111\/trf.12573","article-title":"Efficacy of therapeutic plasma exchange for treatment of stiff-person syndrome","volume":"54","author":"Pagano","year":"2014","journal-title":"Transfusion"},{"key":"ref27","doi-asserted-by":"publisher","first-page":"671","DOI":"10.1046\/j.1537-2995.1999.39070671.x","article-title":"Selective therapeutic extraction of plasma constituents, revisited","volume":"39","author":"Pineda","year":"1999","journal-title":"Transfusion"},{"key":"ref28","doi-asserted-by":"publisher","first-page":"166","DOI":"10.1186\/s40478-025-02082-0","article-title":"Binding patterns of glycine receptor autoantibodies are related to clinical syndromes","volume":"13","author":"Piro","year":"2025","journal-title":"Acta Neuropathol. Commun."},{"key":"ref29","doi-asserted-by":"publisher","first-page":"1731","DOI":"10.1093\/brain\/awaa104","article-title":"Distinctive binding properties of human monoclonal LGI1 autoantibodies determine pathogenic mechanisms","volume":"143","author":"Ramberger","year":"2020","journal-title":"Brain"},{"key":"ref30","doi-asserted-by":"publisher","first-page":"1089101","DOI":"10.3389\/fnmol.2023.1089101","article-title":"Glycine receptor autoantibody binding to the extracellular domain is independent from receptor glycosylation","volume":"16","author":"Rauschenberger","year":"2023","journal-title":"Front. Mol. Neurosci."},{"key":"ref31","doi-asserted-by":"publisher","first-page":"544","DOI":"10.1002\/ana.25832","article-title":"Glycine receptor autoantibodies impair receptor function and induce motor dysfunction","volume":"88","author":"Rauschenberger","year":"2020","journal-title":"Ann. Neurol."},{"key":"ref32","doi-asserted-by":"publisher","first-page":"297","DOI":"10.1016\/j.jns.2016.11.066","article-title":"Stiff person syndrome masquerading as multiple sclerosis","volume":"372","author":"Sabatino","year":"2017","journal-title":"J. Neurol. Sci."},{"key":"ref33","doi-asserted-by":"publisher","first-page":"422","DOI":"10.1523\/JNEUROSCI.1509-14.2015","article-title":"Disturbed neuronal ER-Golgi sorting of unassembled Glycine receptors suggests altered subcellular processing is a cause of human Hyperekplexia","volume":"35","author":"Schaefer","year":"2015","journal-title":"J. Neurosci."},{"key":"ref34","doi-asserted-by":"publisher","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: an open-source platform for biological-image analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods"},{"key":"ref35","doi-asserted-by":"publisher","first-page":"879","DOI":"10.1002\/jcc.20229","article-title":"3D-epitope-explorer (3DEX): localization of conformational epitopes within three-dimensional structures of proteins","volume":"26","author":"Schreiber","year":"2005","journal-title":"J. Comput. Chem."},{"key":"ref36","doi-asserted-by":"publisher","first-page":"219","DOI":"10.1186\/s12974-016-0678-4","article-title":"Mapping autoantigen epitopes: molecular insights into autoantibody-associated disorders of the nervous system","volume":"13","author":"Sinmaz","year":"2016","journal-title":"J. Neuroinflammation"},{"key":"ref37","doi-asserted-by":"publisher","first-page":"e35296","DOI":"10.1371\/journal.pone.0035296","article-title":"Autoantibody epitope spreading in the pre-clinical phase predicts progression to rheumatoid arthritis","volume":"7","author":"Sokolove","year":"2012","journal-title":"PLoS One"},{"key":"ref38","doi-asserted-by":"publisher","first-page":"1081","DOI":"10.3389\/fimmu.2020.01081","article-title":"Innate immune sensing by cells of the adaptive immune system","volume":"11","author":"Stogerer","year":"2020","journal-title":"Front. Immunol."},{"key":"ref39","doi-asserted-by":"publisher","first-page":"1290","DOI":"10.1111\/ene.13721","article-title":"Antiglycine receptor antibody related disease: a case series and literature review","volume":"25","author":"Swayne","year":"2018","journal-title":"Eur. J. Neurol."},{"key":"ref40","doi-asserted-by":"publisher","first-page":"1329013","DOI":"10.3389\/fimmu.2024.1329013","article-title":"Molecular dissection of an immunodominant epitope in K(v)1.2-exclusive autoimmunity","volume":"15","author":"Talucci","year":"2024","journal-title":"Front. Immunol."},{"key":"ref41","doi-asserted-by":"publisher","first-page":"768","DOI":"10.1016\/j.tips.2024.07.007","article-title":"Epitope landscape in autoimmune neurological disease and beyond","volume":"45","author":"Talucci","year":"2024","journal-title":"Trends Pharmacol. Sci."},{"key":"ref42","doi-asserted-by":"publisher","first-page":"36128","DOI":"10.1074\/jbc.M109.043448","article-title":"Mapping of disulfide bonds within the amino-terminal extracellular domain of the inhibitory glycine receptor","volume":"284","author":"Vogel","year":"2009","journal-title":"J. Biol. Chem."},{"key":"ref43","doi-asserted-by":"publisher","first-page":"3962","DOI":"10.1002\/cne.23139","article-title":"Distribution of the glycine receptor beta-subunit in the mouse CNS as revealed by a novel monoclonal antibody","volume":"520","author":"Weltzien","year":"2012","journal-title":"J. Comp. Neurol."},{"key":"ref44","doi-asserted-by":"publisher","first-page":"e0837232023","DOI":"10.1523\/JNEUROSCI.0837-23.2023","article-title":"Role of the Glycine receptor beta subunit in synaptic localization and pathogenicity in severe startle disease","volume":"44","author":"Wiessler","year":"","journal-title":"J. Neurosci."},{"key":"ref45","doi-asserted-by":"publisher","first-page":"e200187","DOI":"10.1212\/NXI.0000000000200187","article-title":"Glycine receptor beta-targeting autoantibodies contribute to the pathology of autoimmune diseases Neurol","volume":"11","author":"Wiessler","year":"","journal-title":"Neuroimmunol. Neuroinflamm"},{"key":"ref46","doi-asserted-by":"publisher","first-page":"e200364","DOI":"10.1212\/NXI.0000000000200364","article-title":"Impaired presynaptic function contributes significantly to the pathology of Glycine receptor autoantibodies","volume":"12","author":"Wiessler","year":"2025","journal-title":"Neurol. Neuroimmunol. Neuroinflamm."},{"key":"ref47","doi-asserted-by":"publisher","first-page":"e2706","DOI":"10.1016\/j.neuron.2021.08.019","article-title":"Characterization of the subunit composition and structure of adult human glycine receptors","volume":"109","author":"Yu","year":"2021","journal-title":"Neuron"},{"key":"ref48","doi-asserted-by":"publisher","first-page":"513","DOI":"10.1038\/s41586-021-04022-z","article-title":"Architecture and assembly mechanism of native glycine receptors","volume":"599","author":"Zhu","year":"2021","journal-title":"Nature"}],"container-title":["Frontiers in Molecular Neuroscience"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fnmol.2026.1747209\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,17]],"date-time":"2026-03-17T14:20:18Z","timestamp":1773757218000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fnmol.2026.1747209\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,17]]},"references-count":48,"alternative-id":["10.3389\/fnmol.2026.1747209"],"URL":"https:\/\/doi.org\/10.3389\/fnmol.2026.1747209","relation":{},"ISSN":["1662-5099"],"issn-type":[{"value":"1662-5099","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,17]]},"article-number":"1747209"}}