{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,18]],"date-time":"2026-06-18T15:33:18Z","timestamp":1781796798679,"version":"3.54.5"},"reference-count":14,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T00:00:00Z","timestamp":1760400000000},"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. Immunol."],"abstract":"<jats:sec><jats:title>Background and objectives<\/jats:title><jats:p>Hashimoto's thyroiditis (HT) is a chronic autoimmune thyroid disorder characterized by B lymphocyte dysregulation and the production of autoantibodies. This study aimed to evaluate a targeted B cell depletion strategy by identifying and validating a disease-associated membrane glycoprotein selectively expressed on B cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>B lymphocytes were isolated from peripheral blood samples of 32 individuals with HT and 40 age- and sex-matched healthy controls (HC). Membrane glycoprotein expression was profiled using a 38-lectin microarray, followed by differential analysis via liquid chromatography-tandem mass spectrometry. Tetraspanin-33 (TSPAN33) was identified for further investigation based on its expression pattern. Recombinant TSPAN33 protein was used as the target in a Systematic Evolution of Ligands by Exponential Enrichment (SELEX) process to generate high-affinity DNA aptamers. This aptamer was chemically conjugated with a CD20 monoclonal antibody (rituximab) using bismaleimide crosslinking to generate a bispecific complex with capable of specific binding pathogenic B lymphocytes. Binding specificity was evaluated using confocal microscopy.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The fluorescence intensity of cell membrane glycoproteins and plasma glycoproteins corresponding to lectins MAL-II and PHA-E was significantly higher in HT patients than in HC patients. The cell membrane protein TSPAN33 bound by the above two lectins is highly expressed in HT patients. The binding energy of aptamer 40 (AP40) to TSPAN33 was 33.2510-9M. AP40 could bind specifically to B cells derived from BALL-1, but not to B cells derived from HC. AP40 can be conjugated with rituximab and purified.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The development of a bispecific aptamerantibody conjugate targeting TSPAN33 offers a promising strategy for selective B cell depletion.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2025.1636003","type":"journal-article","created":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T05:26:46Z","timestamp":1760419606000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Agglutinin chip screening of B cell surface biomarkers in Hashimoto\u2019s thyroiditis for therapeutic targeting"],"prefix":"10.3389","volume":"16","author":[{"given":"Jun-ling","family":"Ren","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiao-ming","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,10,14]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.3389\/fendo.2025.1627787","article-title":"Persistent symptoms in euthyroid Hashimoto's thyroiditis: current hypotheses and emerging management strategies","volume":"16","author":"Zhang","year":"2025","journal-title":"Front Endocrinol (Lausanne)"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.3389\/fendo.2025.1576850","article-title":"Vitamin D deficiency in Hashimoto's thyroiditis: mechanisms, immune modulation, and therapeutic implications","volume":"16","author":"Sun","year":"2025","journal-title":"Front Endocrinol (Lausanne)"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1007\/s40618-024-02370-x","article-title":"CD20\u2009+\u2009T lymphocytes in isolated Hashimoto's thyroiditis and type 3 autoimmune polyendocrine syndrome: a pilot study","volume":"47","author":"Stramazzo","year":"2024","journal-title":"J Endocrinol Invest"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1177\/13524585251361330","article-title":"Disease outcomes following lateral switch among different CD20-antibodies in active multiple sclerosis","volume":"31","author":"Axhausen","year":"2025","journal-title":"Mult Scler"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1016\/j.tips.2025.07.013","article-title":"Targeting glycosylation to enhance tumor immunotherapy","volume":"46","author":"Zhu","year":"2025","journal-title":"Trends Pharmacol Sci"},{"key":"B6","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1186\/s13578-023-00995-8","article-title":"The versatile roles of testrapanins in cancer from intracellular signaling to cell-cell communication: cell membrane proteins without ligands","volume":"13","author":"Zhou","year":"2023","journal-title":"Cell Biosci"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1016\/j.clim.2013.08.005","article-title":"TSPAN33 is a novel marker of activated and Malignant B cells","volume":"149","author":"Luu","year":"2013","journal-title":"Clin Immunol"},{"key":"B8","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1111\/sji.12559","article-title":"Tspan33 is Expressed in Transitional and Memory B Cells, but is not Responsible for High ADAM10 Expression","volume":"86","author":"P\u00e9rez-Mart\u00ednez","year":"2017","journal-title":"Scand J Immunol"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.3390\/biom15040583","article-title":"Plasma exosomal proteomics identifies differentially expressed proteins as biomarkers for acute myocardial infarction","volume":"15","author":"Zhou","year":"2025","journal-title":"Biomolecules"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2022.104777","article-title":"The role of tetraspanins pan-cancer","volume":"25","author":"Huang","year":"2022","journal-title":"iScience"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1111\/febs.15216","article-title":"Tetraspanin 33 (TSPAN33) regulates endocytosis and migration of human B lymphocytes by affecting the tension of the plasma membrane","volume":"287","author":"Navarro-Hernandez","year":"2020","journal-title":"FEBS J"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.2174\/0929867325666181008142831","article-title":"Aptamer-based targeted drug delivery systems: current potential and challenges","volume":"27","author":"He","year":"2020","journal-title":"Curr Med Chem"},{"key":"B13","doi-asserted-by":"publisher","first-page":"e2109609","DOI":"10.1002\/adma.202109609","article-title":"Biomimetic nanoerythrosome-coated aptamer-DNA tetrahedron\/maytansine conjugates: pH-responsive and targeted cytotoxicity for HER2-positive breast cancer","volume":"34","author":"Ma","year":"2022","journal-title":"Adv Mater"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1002\/art.42080","article-title":"Aptamer-based screen of neuropsychiatric lupus cerebrospinal fluid reveals potential biomarkers that overlap with the choroid plexus transcriptome","volume":"74","author":"Vanarsa","year":"2022","journal-title":"Arthritis Rheumatol"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1636003\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T05:26:46Z","timestamp":1760419606000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1636003\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,14]]},"references-count":14,"alternative-id":["10.3389\/fimmu.2025.1636003"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2025.1636003","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,14]]},"article-number":"1636003"}}