{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T18:47:59Z","timestamp":1791571679096,"version":"4.3.4"},"reference-count":47,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2021,6,8]],"date-time":"2021-06-08T00:00:00Z","timestamp":1623110400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Histopathology"],"published-print":{"date-parts":[[2021,10]]},"abstract":"<jats:sec><jats:title>Aims<\/jats:title><jats:p>Programmed death\u2010ligand 1 (PD\u2010L1) immunostaining is used to predict which non\u2010small\u2010cell lung cancer (NSCLC) patients will respond best to treatment with programmed cell death protein 1\/PD\u2010L1 inhibitors. PD\u2010L1 immunostaining is sometimes performed on alcohol\u2010fixed cytological specimens instead of on formalin\u2010fixed paraffin\u2010embedded (FFPE) biopsies or resections. We studied whether ethanol prefixation of clots from endobronchial ultrasound\u2010guided transbronchial needle aspiration (EBUS\u2010TBNA) results in diminished PD\u2010L1 immunostaining as compared with formalin fixation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods and results<\/jats:title><jats:p>FFPE cell blocks from EBUS\u2010TBNA specimens of 54 NSCLC patients were identified. For each case, paired samples were available, consisting of clots directly immersed in formalin and clots prefixed in Fixcyt (50% ethanol). Serial sections were immunostained for PD\u2010L1 by use of the standardised SP263 assay and the 22C3 antibody as a laboratory\u2010developed test (LDT). PD\u2010L1 positivity was determined with two cut\u2010offs (1% and 50%). Concordance of PD\u2010L1 positivity between the formalin\u2010fixed (gold standard) and ethanol\u2010prefixed material was assessed. When the 22C3 LDT was used, 30% and 36% of the ethanol\u2010prefixed specimens showed false\u2010negative results at the 1% and 50% cut\u2010offs, respectively (kappa 0.64 and 0.68). When SP263 was used, 22% of the ethanol\u2010prefixed specimens showed false\u2010negative results at the 1% cut\u2010off (kappa 0.67). At the 50% cut\u2010off, concordance was higher (kappa 0.91), with 12% of the ethanol\u2010prefixed specimens showing false\u2010negative results.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Ethanol fixation of EBUS\u2010TBNA specimens prior to formalin fixation can result in a considerable number of false\u2010negative PD\u2010L1 immunostaining results when a 1% cut\u2010off is used and immunostaining is performed with SP263 or the 22C3 LDT. The same applies to use of the 50% cut\u2010off when immunostaining is performed with the 22C3 LDT.<\/jats:p><\/jats:sec>","DOI":"10.1111\/his.14373","type":"journal-article","created":{"date-parts":[[2021,3,27]],"date-time":"2021-03-27T08:51:55Z","timestamp":1616835115000},"page":"480-490","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["False\u2010negative programmed death\u2010ligand 1 immunostaining in ethanol\u2010fixed endobronchial ultrasound\u2010guided transbronchial needle aspiration specimens of non\u2010small\u2010cell lung cancer patients"],"prefix":"10.1111","volume":"79","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3106-5734","authenticated-orcid":false,"given":"Bregje M","family":"Koomen","sequence":"first","affiliation":[{"name":"Department of Pathology University Medical Centre Utrecht Utrecht University Utrecht the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Willem","family":"Vreuls","sequence":"additional","affiliation":[{"name":"Department of Pathology Canisius\u2010Wilhelmina Hospital Nijmegen the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mirthe","family":"de Boer","sequence":"additional","affiliation":[{"name":"Department of Pathology University Medical Centre Utrecht Utrecht University Utrecht the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emma J","family":"de Ruiter","sequence":"additional","affiliation":[{"name":"Department of Pathology University Medical Centre Utrecht Utrecht University Utrecht the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juergen","family":"Hoelters","sequence":"additional","affiliation":[{"name":"Department of Pulmonology Canisius\u2010Wilhelmina Hospital Nijmegen the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aryan","family":"Vink","sequence":"additional","affiliation":[{"name":"Department of Pathology University Medical Centre Utrecht Utrecht University Utrecht the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan M","family":"Willems","sequence":"additional","affiliation":[{"name":"Department of Pathology University Medical Centre Utrecht Utrecht University Utrecht the Netherlands"},{"name":"Department of Pathology and Medical Biology University of Groningen University Medical Centre Groningen Groningen The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2021,6,8]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aar4060"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1507643"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1504627"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1606774"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(15)01281-7"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(16)32517-X"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1709937"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(18)32409-7"},{"issue":"4","key":"e_1_2_9_10_1","first-page":"iv192\u2013iv237","article-title":"Metastatic non\u2010small cell lung cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow\u2010up","volume":"29","author":"Planchard D","year":"2018","journal-title":"Ann. 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