{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,8,27]],"date-time":"2025-08-27T16:15:45Z","timestamp":1756311345461},"reference-count":13,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2015,12,7]],"date-time":"2015-12-07T00:00:00Z","timestamp":1449446400000},"content-version":"vor","delay-in-days":6,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["255318"],"award-info":[{"award-number":["255318"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Veterinary Clinical Pathol"],"published-print":{"date-parts":[[2015,12]]},"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Immunophenotyping of canine and feline lymphoma to determine B\u2010cell or T\u2010cell origin is important for predicting prognosis and for development of treatment protocols. For advanced diagnostic cytology tests that can be performed on smears are required to predict the immunophenotype of lymphomas.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objectives<\/jats:title><jats:p>The aim of this study was to develop a multiple immunofluorescence (<jats:styled-content style=\"fixed-case\">MIF<\/jats:styled-content>) staining method for the determination of lymphocyte immunophenotype in cytologic specimens, and to evaluate its clinical utility.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>B cells and T cells were detected using anti\u2010<jats:styled-content style=\"fixed-case\">CD<\/jats:styled-content>79\u03b1 and anti\u2010<jats:styled-content style=\"fixed-case\">CD<\/jats:styled-content>3 antibodies, respectively, followed by specific fluorescence\u2010labeled secondary antibodies. The <jats:styled-content style=\"fixed-case\">MIF<\/jats:styled-content> staining method was first developed using fresh\u2010frozen sections of normal canine lymph nodes. The optimal fixative, the necessity of antigen retrieval (<jats:styled-content style=\"fixed-case\">AR<\/jats:styled-content>), and the optimal concentration of the antibodies were determined. The <jats:styled-content style=\"fixed-case\">MIF<\/jats:styled-content> method was then applied to smears of normal lymph nodes, and to clinical samples from dogs and cats with lymphoma. The <jats:styled-content style=\"fixed-case\">MIF<\/jats:styled-content> results were compared to genetic clonality results.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>B and T cells were detected based on specific fluorescence in frozen sections, using formalin fixation without <jats:styled-content style=\"fixed-case\">AR<\/jats:styled-content>. Specific fluorescence was also detected in smears from normal lymph nodes and lymphomas, and the immunophenotypes predicted from this <jats:styled-content style=\"fixed-case\">MIF<\/jats:styled-content> staining method completely corresponded to those from genetic clonality analysis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The <jats:styled-content style=\"fixed-case\">MIF<\/jats:styled-content> staining method that we developed in this study effectively distinguished lymphocyte immunophenotypes with high specificity and sensitivity using a single smear sample, and was useful as a diagnostic tool for canine and feline lymphoma.<\/jats:p><\/jats:sec>","DOI":"10.1111\/vcp.12300","type":"journal-article","created":{"date-parts":[[2015,12,8]],"date-time":"2015-12-08T18:58:54Z","timestamp":1449601134000},"page":"580-585","update-policy":"http:\/\/dx.doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Development and application of multiple immunofluorescence staining for diagnostic cytology of canine and feline lymphoma"],"prefix":"10.1111","volume":"44","author":[{"given":"Mariko","family":"Sawa","sequence":"first","affiliation":[{"name":"Laboratory of Veterinary Clinical Pathology Joint Faculty of Veterinary Medicine Kagoshima University Kagoshima Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akira","family":"Yabuki","sequence":"additional","affiliation":[{"name":"Laboratory of Veterinary Clinical Pathology Joint Faculty of Veterinary Medicine Kagoshima University Kagoshima Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Asuka","family":"Setoguchi","sequence":"additional","affiliation":[{"name":"Laboratory of Small Animal Internal Medicine Joint Faculty of Veterinary Medicine Kagoshima University Kagoshima Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Osamu","family":"Yamato","sequence":"additional","affiliation":[{"name":"Laboratory of Veterinary Clinical Pathology Joint Faculty of Veterinary Medicine Kagoshima University Kagoshima Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2015,12,7]]},"reference":[{"key":"e_1_2_3_2_1","doi-asserted-by":"publisher","DOI":"10.1177\/104063870501700404"},{"key":"e_1_2_3_3_1","first-page":"1179","article-title":"Histologic classification and immunophenotyping of canine non\u2010Hodgkin's lymphomas: unexpected high frequency of T cell lymphomas with B cell morphology","volume":"22","author":"Teske E","year":"1994","journal-title":"Exp Hematol"},{"key":"e_1_2_3_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tvjl.2010.03.011"},{"key":"e_1_2_3_5_1","doi-asserted-by":"publisher","DOI":"10.1354\/vp.40-1-32"},{"key":"e_1_2_3_6_1","first-page":"532","volume-title":"Diagnostic Cytology and Hematology of the Dog and Cat","author":"Barger AM","year":"2014"},{"key":"e_1_2_3_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2011.12.019"},{"key":"e_1_2_3_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2011.12.015"},{"key":"e_1_2_3_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2011.05.030"},{"key":"e_1_2_3_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2005.10.009"},{"key":"e_1_2_3_11_1","doi-asserted-by":"publisher","DOI":"10.1354\/vp.42-5-596"},{"key":"e_1_2_3_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2006.11.005"},{"key":"e_1_2_3_13_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1939-165X.2009.00119.x"},{"key":"e_1_2_3_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.rvsc.2012.03.013"}],"container-title":["Veterinary Clinical Pathology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fvcp.12300","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/vcp.12300","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,24]],"date-time":"2023-09-24T09:45:23Z","timestamp":1695548723000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/vcp.12300"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,12]]},"references-count":13,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2015,12]]}},"alternative-id":["10.1111\/vcp.12300"],"URL":"https:\/\/doi.org\/10.1111\/vcp.12300","archive":["Portico"],"relation":{},"ISSN":["0275-6382","1939-165X"],"issn-type":[{"value":"0275-6382","type":"print"},{"value":"1939-165X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,12]]},"assertion":[{"value":"2015-12-07","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}