{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T13:05:10Z","timestamp":1782997510876,"version":"3.54.5"},"reference-count":19,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2008,9,29]],"date-time":"2008-09-29T00:00:00Z","timestamp":1222646400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cytometry Part B Clinical"],"published-print":{"date-parts":[[2009,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background:<\/jats:title><jats:p>Myasthenia gravis (MG) is an autoimmune disease affecting \u223c40,000 patients in the United States. One of the major mechanisms of disease pathology in MG is the binding, internalization, and eventual destruction of acetylcholine receptors (AChR) at the neuromuscular junction by cross\u2010linking AChR\u2010specific autoantibodies. This process, known as antigenic modulation, ultimately attenuates the ability of muscle cells to contract in response to signals from neurons, leading to muscle weakness and fatigue. For this reason, antigenic modulation of the AChR on cultured cells has become an important diagnostic tool for assessing the pathogenicity of AChR\u2010specific autoantibodies. Traditionally, these assays have been done using radiolabeled AChR ligands such as <jats:sup>125<\/jats:sup>I alpha\u2010bungarotoxin to determine relative AChR number. Here, we present a high\u2010throughput immunofluorescent flow cytometry\u2010based assay that can be used to quantify AChR levels on the cell surface and assess the efficacy of molecules designed to rescue antigenic modulation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods:<\/jats:title><jats:p>AChR levels were quantified on human muscle cells before and after treatment with AChR antibodies via immunofluorescent labeling with the AChR monoclonal antibodies, mAb210 and mAb B3, followed by flow cytometry of EDTA\u2010treated cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results:<\/jats:title><jats:p>Using a novel, flow cytometry\u2010based assay, antigenic modulation of the AChR was demonstrated on human cells using both AChR\u2010specific monoclonal antibody and MG patient serum. The degree of antigenic modulation was dose responsive to antibody levels and could be reversed by preincubating antibodies with soluble AChR alpha subunit extracellular domain.<\/jats:p><\/jats:sec><jats:sec><jats:title>Summary:<\/jats:title><jats:p>A rapid, nonradioactive assay was developed to determine the potential of AChR\u2010specific antibodies in the serum of MG patients to bind and down\u2010regulate the AChR. This assay can be used to assess the ability of putative therapeutics that rescue antigenic modulation and could be developed for the treatment of MG. \u00a9 2008 Clinical Cytometry Society<\/jats:p><\/jats:sec>","DOI":"10.1002\/cyto.b.20454","type":"journal-article","created":{"date-parts":[[2008,9,29]],"date-time":"2008-09-29T18:43:23Z","timestamp":1222713803000},"page":"206-212","source":"Crossref","is-referenced-by-count":17,"title":["A rapid, fluorescence\u2010based assay for detecting antigenic modulation of the acetylcholine receptor on human cell lines"],"prefix":"10.1002","volume":"76B","author":[{"given":"Dennis","family":"Keefe","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Donna","family":"Hess","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer","family":"Bosco","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Socrates","family":"Tzartos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jan","family":"Powell","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justin","family":"Lamsa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Serene","family":"Josiah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2008,9,29]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI29894"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.74.7.3090"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/274068a0"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)50366-5"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1002\/mus.880020304"},{"key":"e_1_2_6_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneuroim.2004.10.002"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.1212\/WNL.28.3.266"},{"key":"e_1_2_6_9_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.144.3.726"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/0165-5728(87)90067-1"},{"key":"e_1_2_6_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/0165-5728(87)90092-0"},{"key":"e_1_2_6_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneuroim.2007.06.014"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.1212\/WNL.47.6.1552"},{"key":"e_1_2_6_14_2","doi-asserted-by":"publisher","DOI":"10.1002\/eji.1830220939"},{"key":"e_1_2_6_15_2","doi-asserted-by":"publisher","DOI":"10.1002\/eji.1830230816"},{"key":"e_1_2_6_16_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1521-4141(199802)28:02<616::AID-IMMU616>3.0.CO;2-I"},{"key":"e_1_2_6_17_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110731200"},{"key":"e_1_2_6_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/0165-5728(86)90105-0"},{"key":"e_1_2_6_19_2","doi-asserted-by":"crossref","first-page":"5916","DOI":"10.1016\/S0021-9258(18)60652-0","article-title":"Monoclonal antibodies to the main immunogenic region of the nicotinic acetylcholine receptor bind to residues 61\u201076 of the alpha subunit","volume":"263","author":"Barkas T","year":"1988","journal-title":"J Biol Chem"},{"key":"e_1_2_6_20_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.97.2.761"}],"container-title":["Cytometry Part B: Clinical Cytometry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fcyto.b.20454","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cyto.b.20454","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,11]],"date-time":"2023-10-11T15:52:21Z","timestamp":1697039541000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/cyto.b.20454"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2008,9,29]]},"references-count":19,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2009,5]]}},"alternative-id":["10.1002\/cyto.b.20454"],"URL":"https:\/\/doi.org\/10.1002\/cyto.b.20454","archive":["Portico"],"relation":{},"ISSN":["1552-4949","1552-4957"],"issn-type":[{"value":"1552-4949","type":"print"},{"value":"1552-4957","type":"electronic"}],"subject":[],"published":{"date-parts":[[2008,9,29]]}}}