{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T07:16:20Z","timestamp":1791270980084,"version":"4.1.0"},"reference-count":74,"publisher":"American Society for Clinical Investigation","issue":"12","license":[{"start":{"date-parts":[[2019,6,20]],"date-time":"2019-06-20T00:00:00Z","timestamp":1560988800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100005202","name":"Muscular Dystrophy Association","doi-asserted-by":"crossref","award":["MDA575198"],"award-info":[{"award-number":["MDA575198"]}],"id":[{"id":"10.13039\/100005202","id-type":"DOI","asserted-by":"crossref"}]},{"name":"NIAID NIH","award":["R01AI114780"],"award-info":[{"award-number":["R01AI114780"]}]},{"DOI":"10.13039\/100000060","name":"NIAID","doi-asserted-by":"crossref","award":["1R21AI142198"],"award-info":[{"award-number":["1R21AI142198"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Conquer Myasthenia Gravis","award":["pilot research award"],"award-info":[{"award-number":["pilot research award"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2019,6,20]]},"abstract":"<jats:p>Myasthenia gravis (MG) is a chronic autoimmune disorder characterized by muscle weakness and caused by pathogenic autoantibodies that bind to membrane proteins at the neuromuscular junction. Most patients have autoantibodies against the acetylcholine receptor (AChR), but a subset of patients have autoantibodies against muscle-specific tyrosine kinase (MuSK) instead. MuSK is an essential component of the pathway responsible for synaptic differentiation, which is activated by nerve-released agrin. Through binding MuSK, serum-derived autoantibodies inhibit agrin-induced MuSK autophosphorylation, impair clustering of AChRs, and block neuromuscular transmission. We sought to establish individual MuSK autoantibody clones so that the autoimmune mechanisms could be better understood. We isolated MuSK autoantibody-expressing B cells from 6 MuSK MG patients using a fluorescently tagged MuSK antigen multimer, then generated a panel of human monoclonal autoantibodies (mAbs) from these cells. Here we focused on 3 highly specific mAbs that bound quantitatively to MuSK in solution, to MuSK-expressing HEK cells, and at mouse neuromuscular junctions, where they colocalized with AChRs. These 3 IgG isotype mAbs (2 IgG4 and 1 IgG3 subclass) recognized the Ig-like domain 2 of MuSK. The mAbs inhibited AChR clustering, but intriguingly, they enhanced rather than inhibited MuSK phosphorylation, which suggests an alternative mechanism for inhibiting AChR clustering.<\/jats:p>","DOI":"10.1172\/jci.insight.127167","type":"journal-article","created":{"date-parts":[[2019,6,19]],"date-time":"2019-06-19T11:01:08Z","timestamp":1560942068000},"source":"Crossref","is-referenced-by-count":73,"title":["Characterization of pathogenic monoclonal autoantibodies derived from muscle-specific kinase myasthenia gravis patients"],"prefix":"10.1172","volume":"4","author":[{"given":"Kazushiro","family":"Takata","sequence":"first","affiliation":[{"name":"Department of Neurology and"},{"name":"Department of Immunobiology, Yale School of Medicine, Yale University, New Haven, Connecticut, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Panos","family":"Stathopoulos","sequence":"additional","affiliation":[{"name":"Department of Neurology and"},{"name":"Department of Immunobiology, Yale School of Medicine, Yale University, New Haven, Connecticut, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michelangelo","family":"Cao","sequence":"additional","affiliation":[{"name":"Neurosciences Group, Weatherall Institute of Molecular Medicine and Nuffield Department of Clinical Neurosciences, Oxford, England."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marina","family":"Man\u00e9-Damas","sequence":"additional","affiliation":[{"name":"Department of Psychiatry and Neuropsychology, School for Mental Health and Neuroscience, Maastricht University, Maastricht, Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miriam L.","family":"Fichtner","sequence":"additional","affiliation":[{"name":"Department of Neurology and"},{"name":"Department of Immunobiology, Yale School of Medicine, Yale University, New Haven, Connecticut, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erik S.","family":"Benotti","sequence":"additional","affiliation":[{"name":"Department of Neurology and"},{"name":"Department of Immunobiology, Yale School of Medicine, Yale University, New Haven, Connecticut, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leslie","family":"Jacobson","sequence":"additional","affiliation":[{"name":"Oxford Autoimmune Neurology Group, Nuffield Department of Clinical Neurosciences, John Radcliffe Hospital, University of Oxford, Oxford, England."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick","family":"Waters","sequence":"additional","affiliation":[{"name":"Oxford Autoimmune Neurology Group, Nuffield Department of Clinical Neurosciences, John Radcliffe Hospital, University of Oxford, Oxford, England."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarosh R.","family":"Irani","sequence":"additional","affiliation":[{"name":"Oxford Autoimmune Neurology Group, Nuffield Department of Clinical Neurosciences, John Radcliffe Hospital, University of Oxford, Oxford, England."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5990-3373","authenticated-orcid":true,"given":"Pilar","family":"Martinez-Martinez","sequence":"additional","affiliation":[{"name":"Department of Psychiatry and Neuropsychology, School for Mental Health and Neuroscience, Maastricht University, Maastricht, Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Beeson","sequence":"additional","affiliation":[{"name":"Neurosciences Group, Weatherall Institute of Molecular Medicine and Nuffield Department of Clinical Neurosciences, Oxford, England."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9761-0002","authenticated-orcid":true,"given":"Mario","family":"Losen","sequence":"additional","affiliation":[{"name":"Department of Psychiatry and Neuropsychology, School for Mental Health and Neuroscience, Maastricht University, Maastricht, Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angela","family":"Vincent","sequence":"additional","affiliation":[{"name":"Neurosciences Group, Weatherall Institute of Molecular Medicine and Nuffield Department of Clinical Neurosciences, Oxford, England."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard J.","family":"Nowak","sequence":"additional","affiliation":[{"name":"Department of Neurology and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kevin C.","family":"O\u2019Connor","sequence":"additional","affiliation":[{"name":"Department of Neurology and"},{"name":"Department of Immunobiology, Yale School of Medicine, Yale University, New Haven, Connecticut, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"232","published-online":{"date-parts":[[2019,6,20]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1038\/nri916"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMra1602678"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1002\/mus.25973"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1033.2000.01785.x"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1084\/jem.144.3.739"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1172\/JCI66039"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1038\/85520"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM197805182982004"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaut.2013.12.004"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1002\/ana.22312"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1136\/jnnp-2012-303664"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1016\/j.nmd.2016.04.004"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1007\/s13760-014-0364-1"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1002\/ana.20061"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2222.2009.03207.x"},{"issue":"pt 4","key":"B16","doi-asserted-by":"crossref","first-page":"1081","DOI":"10.1093\/brain\/aws025","article-title":"Muscle-specific kinase myasthenia gravis IgG4 autoantibodies cause severe neuromuscular junction dysfunction in mice","volume":"135","author":"Klooster","year":"2012","journal-title":"Brain"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1016\/j.expneurol.2012.01.025"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1172\/JCI21545"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.273.11.6467"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1313944110"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0080695"},{"key":"B22","doi-asserted-by":"crossref","DOI":"10.1038\/srep13928","article-title":"Collagen Q and anti-MuSK autoantibody competitively suppress agrin\/LRP4\/MuSK signaling","volume":"5","author":"Otsuka","year":"2015","journal-title":"Sci Rep"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaut.2016.11.005"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1126\/science.1144603"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-01019-5"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1212\/WNL.0b013e3182407982"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1177\/1756285611411503"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2014-02-555169"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1002\/art.11181"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1038\/jid.2013.236"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1111\/nyas.13535"},{"issue":"17","key":"B32","doi-asserted-by":"crossref","DOI":"10.1172\/jci.insight.94263","article-title":"Autoantibody-producing plasmablasts after B cell depletion identified in muscle-specific kinase myasthenia gravis","volume":"2","author":"Stathopoulos","year":"2017","journal-title":"JCI Insight"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2011-03-341917"},{"issue":"pt 10","key":"B34","doi-asserted-by":"crossref","first-page":"2304","DOI":"10.1093\/brain\/awg223","article-title":"Clinical correlates with anti-MuSK antibodies in generalized seronegative myasthenia gravis","volume":"126","author":"Evoli","year":"2003","journal-title":"Brain"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1212\/01.wnl.0000254620.45529.97"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1212\/01.WNL.0000065882.63904.53"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1833158100"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2006.09.019"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2008.10.002"},{"issue":"175","key":"B40","doi-asserted-by":"crossref","DOI":"10.1126\/scitranslmed.3005166","article-title":"Long-term remissions of severe pemphigus after rituximab therapy are associated with prolonged failure of desmoglein B cell response","volume":"5","author":"Colliou","year":"2013","journal-title":"Sci Transl Med"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1093\/brain\/awy010"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1002\/ana.25173"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1172\/JCI64413"},{"key":"B44","doi-asserted-by":"crossref","DOI":"10.1038\/ncomms5167","article-title":"Shared VH1-46 gene usage by pemphigus vulgaris autoantibodies indicates common humoral immune responses among patients","volume":"5","author":"Cho","year":"2014","journal-title":"Nat Commun"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0185976"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1212\/NXI.0000000000000357"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1212\/NXI.0000000000000089"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2009.07.091"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneuroim.2015.12.016"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.1101\/gad.279745.116"},{"key":"B51","doi-asserted-by":"crossref","DOI":"10.7554\/eLife.34375.018","article-title":"Preserving neuromuscular synapses in ALS by stimulating MuSK with a therapeutic agonist antibody","author":"Cantor","year":"2018","journal-title":"Elife"},{"issue":"1\u20132","key":"B52","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/j.jneuroim.2008.01.013","article-title":"Clinical fluctuations in MuSK myasthenia gravis are related to antigen-specific IgG4 instead of IgG1","volume":"195","author":"Niks","year":"2008","journal-title":"J Neuroimmunol"},{"key":"B53","doi-asserted-by":"publisher","DOI":"10.1212\/NXI.0000000000000547"},{"issue":"1\u20132","key":"B54","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jneuroim.2011.12.005","article-title":"Divalent and monovalent autoantibodies cause dysfunction of MuSK by distinct mechanisms in a rabbit model of myasthenia gravis","volume":"244","author":"Mori","year":"2012","journal-title":"J Neuroimmunol"},{"issue":"2","key":"B55","doi-asserted-by":"crossref","first-page":"743","DOI":"10.4049\/jimmunol.163.2.743","article-title":"I-Ag7-mediated antigen presentation by B lymphocytes is critical in overcoming a checkpoint in T cell tolerance to islet beta cells of nonobese diabetic mice","volume":"163","author":"Noorchashm","year":"1999","journal-title":"J Immunol"},{"issue":"8","key":"B56","doi-asserted-by":"crossref","first-page":"3912","DOI":"10.4049\/jimmunol.161.8.3912","article-title":"B lymphocytes are critical antigen-presenting cells for the initiation of T cell-mediated autoimmune diabetes in nonobese diabetic mice","volume":"161","author":"Serreze","year":"1998","journal-title":"J Immunol"},{"key":"B57","doi-asserted-by":"publisher","DOI":"10.1038\/35100573"},{"key":"B58","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20130699"},{"key":"B59","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1402236"},{"key":"B60","doi-asserted-by":"publisher","DOI":"10.1007\/s00281-014-0424-x"},{"key":"B61","doi-asserted-by":"publisher","DOI":"10.1136\/ard.2008.096495"},{"key":"B62","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaf6756"},{"key":"B63","doi-asserted-by":"publisher","DOI":"10.1002\/acn3.311"},{"key":"B64","doi-asserted-by":"publisher","DOI":"10.1016\/0165-5728(84)90064-X"},{"key":"B65","doi-asserted-by":"publisher","DOI":"10.1002\/ana.21641"},{"issue":"4","key":"B66","doi-asserted-by":"crossref","first-page":"1919","DOI":"10.4049\/jimmunol.158.4.1919","article-title":"Human anti-nicotinic acetylcholine receptor recombinant Fab fragments isolated from thymus-derived phage display libraries from myasthenia gravis patients reflect predominant specificities in serum and block the action of pathogenic serum antibodies","volume":"158","author":"Graus","year":"1997","journal-title":"J Immunol"},{"key":"B67","doi-asserted-by":"publisher","DOI":"10.1093\/brain\/awz106"},{"key":"B68","doi-asserted-by":"publisher","DOI":"10.1002\/hep.26232"},{"key":"B69","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0111726"},{"issue":"Pt 7","key":"B70","doi-asserted-by":"crossref","first-page":"1940","DOI":"10.1093\/brain\/awn092","article-title":"IgG1 antibodies to acetylcholine receptors in \u2018seronegative\u2019 myasthenia gravis","volume":"131","author":"Leite","year":"2008","journal-title":"Brain"},{"issue":"Web Server issue","key":"B71","doi-asserted-by":"crossref","first-page":"W503","DOI":"10.1093\/nar\/gkn316","article-title":"IMGT\/V-QUEST: the highly customized and integrated system for IG and TR standardized V-J and V-D-J sequence analysis","volume":"36","author":"Brochet","year":"2008","journal-title":"Nucleic Acids Res"},{"key":"B72","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/29.1.207"},{"issue":"94","key":"B73","doi-asserted-by":"crossref","DOI":"10.3791\/52220-v","article-title":"The neuromuscular junction: measuring synapse size, fragmentation and changes in synaptic protein density using confocal fluorescence microscopy","author":"Tse","year":"2014","journal-title":"J Vis Exp"},{"key":"B74","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajpath.2016.05.025"}],"container-title":["JCI Insight"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/insight.jci.org\/articles\/view\/127167\/files\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T12:28:38Z","timestamp":1779193718000},"score":1,"resource":{"primary":{"URL":"https:\/\/insight.jci.org\/articles\/view\/127167"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,6,20]]},"references-count":74,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2019,6,20]]},"published-print":{"date-parts":[[2019,6,20]]}},"alternative-id":["10.1172\/jci.insight.127167"],"URL":"https:\/\/doi.org\/10.1172\/jci.insight.127167","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/488247","asserted-by":"object"}]},"ISSN":["2379-3708"],"issn-type":[{"value":"2379-3708","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,6,20]]},"article-number":"e127167"}}