{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T02:12:44Z","timestamp":1775009564988,"version":"3.50.1"},"reference-count":41,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2024,5,6]],"date-time":"2024-05-06T00:00:00Z","timestamp":1714953600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100008723","name":"Suomen L\u00e4\u00e4ketieteen S\u00e4\u00e4ti\u00f6","doi-asserted-by":"publisher","award":["5954"],"award-info":[{"award-number":["5954"]}],"id":[{"id":"10.13039\/100008723","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100015738","name":"Mary och Georg C. Ehrnrooths Stiftelse","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100015738","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100015738","name":"Mary och Georg C. Ehrnrooths Stiftelse","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100015738","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008413","name":"Instrumentariumin Tiedes\u00e4\u00e4ti\u00f6","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100008413","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Finnish State Research Funding \/ Valtion tutkimusrahoitus"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Toll-like receptors (TLRs) are pattern recognition receptors of the innate immunity. TLRs are known to mediate both antitumor effects and tumorigenesis. TLRs are abundant in many cancers, but their expression in small bowel neuroendocrine tumors (SB-NETs) is unknown. We aimed to characterize the expression of TLRs 1\u20139 in SB-NETs and lymph node metastases and evaluate their prognostic relevance. The present study included 125 patients with SB-NETs, of whom 95 had lymph node metastases, from two Finnish hospitals. Tissue samples were stained immunohistochemically for TLR expression, assessed based on cytoplasmic and nucleic staining intensity and percentage of positively stained cells. Statistical methods for survival analysis included Kaplan-Meier method and Cox regression adjusted for confounding factors. Disease-specific survival (DSS) was the primary outcome. TLRs 1\u20132 and 4\u20139 were expressed in SB-NETs and lymph node metastases. TLR3 showed no positive staining. In primary SB-NETs, TLRs 1\u20139 were not associated with survival. For lymph node metastases, high cytoplasmic TLR7 intensity associated with worse DSS compared to low cytoplasmic intensity (26.4% vs. 84.9%, p = 0.028). Adjusted mortality hazard (HR) was 3.90 (95% CI 1.07\u201314.3). The expression of TLRs 1\u20136 and 8\u20139 in lymph node metastases were not associated with survival. SB-NETs and their lymph node metastases express cytoplasmic TLR 1\u20132 and 4\u20139 and nucleic TLR5. High TLR7 expression in SB-NET lymph node metastases was associated with worse prognosis. The current research has future perspective, as it can help create base for clinical drug trials to target specific TLRs with agonists or antagonists to treat neuroendocrine tumors.<\/jats:p>","DOI":"10.1371\/journal.pone.0302813","type":"journal-article","created":{"date-parts":[[2024,5,6]],"date-time":"2024-05-06T17:49:28Z","timestamp":1715017768000},"page":"e0302813","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["Toll-like receptors 1\u20139 in small bowel neuroendocrine tumors\u2013Clinical significance and 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Takeuchi","year":"1999","journal-title":"Immunity"},{"issue":"6857","key":"pone.0302813.ref012","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1038\/35099560","article-title":"Recognition of double-stranded RNA and activation of NF-kappaB by Toll-like receptor 3","volume":"413","author":"L Alexopoulou","year":"2001","journal-title":"Nature"},{"issue":"5396","key":"pone.0302813.ref013","first-page":"2085","article-title":"Defective LPS Signaling in C3H\/HeJ and C57BL\/10ScCr Mice: Mutations in Tlr4","volume":"282","author":"A Poltorak","year":"1998","journal-title":"Gene. 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Hiltunen","year":"2021","journal-title":"Diseases"},{"key":"pone.0302813.ref025","author":"E Woltering","year":"2017","edition":"8"},{"issue":"2","key":"pone.0302813.ref026","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1111\/his.13975","article-title":"The 2019 WHO classification of tumours of the digestive system.","volume":"76","author":"ID Nagtegaal","year":"2020","journal-title":"Histopathology."},{"issue":"1","key":"pone.0302813.ref027","doi-asserted-by":"crossref","DOI":"10.1038\/s41598-017-17204-5","article-title":"QuPath: Open source software for digital pathology image analysis.","volume":"7","author":"P Bankhead","year":"2017","journal-title":"Sci Rep."},{"issue":"4","key":"pone.0302813.ref028","first-page":"1843","article-title":"The Expression of Toll-like Receptors 2, 4, 5, 7 and 9 in Merkel Cell Carcinoma","volume":"35","author":"L Jouhi","year":"2015","journal-title":"Anticancer 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Young","year":"2021","journal-title":"Gut."},{"issue":"4","key":"pone.0302813.ref032","doi-asserted-by":"crossref","first-page":"676","DOI":"10.16899\/jcm.1315131","article-title":"Association of Toll-Like Receptor 9 Expression With Prognosis in Breast Carcinoma.","volume":"13","author":"ZE \u00c7elik","year":"2023","journal-title":"J Contemp Med."},{"issue":"4","key":"pone.0302813.ref033","doi-asserted-by":"crossref","first-page":"1285","DOI":"10.1172\/JCI36551","article-title":"Triggering of TLR7 and TLR8 expressed by human lung cancer cells induces cell survival and chemoresistance","volume":"120","author":"J Cherfils-Vicini","year":"2010","journal-title":"J Clin Invest"},{"key":"pone.0302813.ref034","doi-asserted-by":"crossref","first-page":"154673","DOI":"10.1016\/j.prp.2023.154673","article-title":"Implications of Toll-like receptors (TLRs) and their signaling mechanisms in human cancers.","volume":"248","author":"SA Chandrasekar","year":"2023","journal-title":"Pathol Res 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