{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T11:32:14Z","timestamp":1784633534266,"version":"3.55.0"},"reference-count":30,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,4,13]],"date-time":"2021-04-13T00:00:00Z","timestamp":1618272000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2021,4,13]],"date-time":"2021-04-13T00:00:00Z","timestamp":1618272000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100007917","name":"Agricultural Research Service","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100007917","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Vet Res"],"published-print":{"date-parts":[[2021,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Cell-mediated immune responses to <jats:italic>Mycobacterium avium<\/jats:italic> subsp. <jats:italic>paratuberculosis<\/jats:italic> (MAP) are regulated by various types of T lymphocytes. The aim of this study was to quantitate T cell subsets in the mid-ileum of cows naturally infected with MAP to identify differences during different stages of infection, and to determine whether these subsets could be used as predictors of disease state. Immunofluorescent labeling of T cell subsets and macrophages was performed on frozen mid-ileal tissue sections archived from naturally infected dairy cows in either subclinical or clinical disease status, and noninfected control cows. Comprehensive IF staining for CD4, CD8\u03b1, TcR1-N24 (gamma delta), FoxP3, CXCR3 and CCR9 served to define T cell subsets and was correlated with macrophages present. Clinically affected cows demonstrated significantly higher numbers of CXCR3<jats:sup>+<\/jats:sup> (Th1-type) and CCR9<jats:sup>+<\/jats:sup> (total small intestinal lymphocytes) cells at the site of infection compared to the subclinical cows and noninfected controls. Further, predictive modeling indicated a significant interaction between CXCR3<jats:sup>+<\/jats:sup> and AM3K<jats:sup>+<\/jats:sup> (macrophages) cells, suggesting that progression to clinical disease state aligns with increased numbers of these cell types at the site of infection. The ability to predict disease state with this model was improved from previous modeling using immunofluorescent macrophage data. Predictive modelling indicated an interaction between CXCR3<jats:sup>+<\/jats:sup> and AM3K<jats:sup>+<\/jats:sup> cells, which could more sensitively detect subclinical cows compared to clinical cows. It may be possible to use this knowledge to improve and develop an assay to detect subclinically infected animals with more confidence during the early stages of the disease.<\/jats:p>","DOI":"10.1186\/s13567-021-00925-x","type":"journal-article","created":{"date-parts":[[2021,4,13]],"date-time":"2021-04-13T09:04:09Z","timestamp":1618304649000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Prediction of Johne\u2019s disease state based on quantification of T cell markers and their interaction with macrophages in the bovine intestine"],"prefix":"10.1186","volume":"52","author":[{"given":"Caitlin J.","family":"Jenvey","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adrienne L.","family":"Shircliff","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elsa","family":"Obando Marrero","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9039-8582","authenticated-orcid":false,"given":"Judith R.","family":"Stabel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,4,13]]},"reference":[{"key":"925_CR1","doi-asserted-by":"publisher","first-page":"465","DOI":"10.1016\/S0378-1135(00)00331-X","volume":"77","author":"JR Stabel","year":"2000","unstructured":"Stabel JR (2000) Transitions in immune responses to Mycobacterium paratuberculosis. Vet Microbiol 77:465\u2013473","journal-title":"Vet Microbiol"},{"key":"925_CR2","doi-asserted-by":"publisher","first-page":"214384","DOI":"10.4061\/2011\/214384","volume":"2011","author":"BL Plattner","year":"2011","unstructured":"Plattner BL, Hostetter JM (2011) Comparative gamma delta T cell immunology: a focus on mycobacterial disease in cattle. Vet Med Int 2011:214384. https:\/\/doi.org\/10.4061\/2011\/214384","journal-title":"Vet Med Int"},{"key":"925_CR3","doi-asserted-by":"publisher","first-page":"3856","DOI":"10.1128\/IAI.70.7.3856-3864.2002","volume":"70","author":"A Koets","year":"2002","unstructured":"Koets A, Rutten V, Hoek A, van Mil F, Muller K, Bakker D, Gruys E, van Eden W (2002) Progressive bovine paratuberculosis is associated with local loss of CD4+ T cells, increased frequency of gamma delta T cells, and related changes in T-cells function. Infect Immun 70:3856\u20133864. https:\/\/doi.org\/10.1128\/IAI.70.7.3856-3864.2002","journal-title":"Infect Immun"},{"key":"925_CR4","doi-asserted-by":"publisher","first-page":"234","DOI":"10.1016\/j.vetimm.2008.05.019","volume":"125","author":"DE de Almeida","year":"2008","unstructured":"de Almeida DE, Colvin CJ, Coussens PM (2008) Antigen-specific regulatory T cells in bovine paratuberculosis. Vet Immunol Immunopathol 125:234\u2013245. https:\/\/doi.org\/10.1016\/j.vetimm.2008.05.019","journal-title":"Vet Immunol Immunopathol"},{"key":"925_CR5","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1016\/j.vetimm.2016.03.008","volume":"181","author":"JA Roussey","year":"2016","unstructured":"Roussey JA, Oliveira LJ, Langohr IM, Sledge DG, Coussens PM (2016) Regulatory T cells and immune profiling in johne\u2019s disease lesions. Vet Immunol Immunopathol 181:39\u201350. https:\/\/doi.org\/10.1016\/j.vetimm.2016.03.008","journal-title":"Vet Immunol Immunopathol"},{"key":"925_CR6","doi-asserted-by":"publisher","first-page":"620","DOI":"10.1016\/j.yexcr.2010.12.017","volume":"317","author":"JR Groom","year":"2011","unstructured":"Groom JR, Luster AD (2011) CXCR3 in T cell function. Exp Cell Res 317:620\u2013631. https:\/\/doi.org\/10.1016\/j.yexcr.2010.12.017","journal-title":"Exp Cell Res"},{"key":"925_CR7","doi-asserted-by":"publisher","first-page":"1241","DOI":"10.1084\/jem.190.9.1241","volume":"190","author":"BA Zabel","year":"1999","unstructured":"Zabel BA, Agace WW, Campbell JJ, Heath HM, Parent D, Roberts AI, Ebert EC, Kassam N, Qin S, Zovko M, LaRosa GJ, Yang L-L, Soler D, Butcher EC, Ponath PD, Parker CM, Andrew DP (1999) Human G protein-coupled receptor GPR-9-6\/CC chemokine receptor 9 is selectively expressed on intestinal homing T lymphocytes, mucosal lymphocytes, and thymocytes and is required for thymus-expressed chemokine-mediated chemotaxis. J Exp Med 190:1241\u20131256. https:\/\/doi.org\/10.1084\/jem.190.9.1241","journal-title":"J Exp Med"},{"key":"925_CR8","doi-asserted-by":"publisher","first-page":"6296","DOI":"10.3168\/jds.2014-8204","volume":"97","author":"JR Stabel","year":"2014","unstructured":"Stabel JR, Bradner L, Robbe-Austerman S, Beitz DC (2014) Clinical disease and stage of lactation influence shedding of Mycobacterium avium subspecies paratuberculosis into milk and colostrum of naturally infected dairy cows. J Dairy Sci 97:6296\u20136304. https:\/\/doi.org\/10.3168\/jds.2014-8204","journal-title":"J Dairy Sci"},{"key":"925_CR9","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1016\/S0165-2427(01)00253-7","volume":"79","author":"J Stabel","year":"2001","unstructured":"Stabel J, Whitlock R (2001) An evaluation of a modified interferon-gamma assay for the detection of paratuberculosis in dairy herds. Vet Immunol Immunopathol 79:69\u201381","journal-title":"Vet Immunol Immunopathol"},{"key":"925_CR10","unstructured":"R Core Team (2019) R: a language and environment for statistical computing. R Foundation for Statistical Computing. https:\/\/www.R-project.org\/"},{"key":"925_CR11","first-page":"507","volume":"54","author":"R Rigby","year":"2005","unstructured":"Rigby R, Stasinopoulos D (2005) Generalized additive models for location, scale and shape. Appl Stat 54:507\u2013554","journal-title":"Appl Stat"},{"key":"925_CR12","unstructured":"Barton K (2009) Mu-MIn: multi-model inference. http:\/\/R-Forge.R-project.org\/projects\/mumin\/"},{"key":"925_CR13","unstructured":"al ASem (2020) DescTools: tools for descriptive statistics. https:\/\/cran.r-project.org\/package=DescTools"},{"key":"925_CR14","volume-title":"An R companion to applied regression","author":"J Fox","year":"2019","unstructured":"Fox J, Weisberg S (2019) An R companion to applied regression, 3rd edn. Sage, Thousand Oaks","edition":"3"},{"key":"925_CR15","doi-asserted-by":"publisher","first-page":"1","DOI":"10.18637\/jss.v087.i09","volume":"87","author":"J Fox","year":"2018","unstructured":"Fox J, Weisberg S (2018) Visualizing fit and lack of fit in complex regression models with predictor effect plots and partial residuals. J Stat Softw 87:1\u201327","journal-title":"J Stat Softw"},{"key":"925_CR16","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-319-24277-4","volume-title":"ggplot2: elegant graphics for data analysis","author":"H Wickham","year":"2016","unstructured":"Wickham H (2016) ggplot2: elegant graphics for data analysis. Springer-Verlag, New York"},{"key":"925_CR17","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1016\/j.vetimm.2018.06.012","volume":"202","author":"CJ Jenvey","year":"2018","unstructured":"Jenvey CJ, Hostetter JM, Shircliff AL, Stabel JR (2018) Relationship between the pathology of bovine intestinal tissue and current diagnostic tests for Johne\u2019s disease. Vet Immunol Immunopathol 202:93\u2013101. https:\/\/doi.org\/10.1016\/j.vetimm.2018.06.012","journal-title":"Vet Immunol Immunopathol"},{"key":"925_CR18","doi-asserted-by":"publisher","first-page":"e00650-00619","DOI":"10.1128\/IAI.00650-19","volume":"88","author":"JR Stabel","year":"2019","unstructured":"Stabel JR, Bannantine JP (2019) Divergent antigen-specific cellular immune responses during asymptomatic subclinical and clinical states of disease in cows naturally infected with Mycobacterium avium subsp. paratuberculosis. Infect Immun 88:e00650-00619. https:\/\/doi.org\/10.1128\/IAI.00650-19","journal-title":"Infect Immun"},{"key":"925_CR19","doi-asserted-by":"publisher","first-page":"671","DOI":"10.1177\/0300985819844823","volume":"56","author":"CJ Jenvey","year":"2019","unstructured":"Jenvey CJ, Hostetter JM, Shircliff AL, Bannantine JP, Stabel JR (2019) Quantification of macrophages and mycobacterium avium subsp. paratuberculosis in bovine intestinal tissue during different stages of Johne\u2019s disease. Vet Pathol 56:671\u2013680. https:\/\/doi.org\/10.1177\/0300985819844823","journal-title":"Vet Pathol"},{"key":"925_CR20","doi-asserted-by":"publisher","first-page":"e0217649","DOI":"10.1371\/journal.pone.0217649","volume":"14","author":"CJ Jenvey","year":"2019","unstructured":"Jenvey CJ, Shircliff AL, Bannantine JP, Stabel JR (2019) Phenotypes of macrophages present in the intestine are impacted by stage of disease in cattle naturally infected with Mycobacterium avium subsp. paratuberculosis. PLoS One 14:e0217649. https:\/\/doi.org\/10.1371\/journal.pone.0217649","journal-title":"PLoS ONE"},{"key":"925_CR21","doi-asserted-by":"publisher","first-page":"101422","DOI":"10.1016\/j.cimid.2020.101422","volume":"69","author":"M Criado","year":"2020","unstructured":"Criado M, Benavides J, Vallejo R, Arteche N, Guti\u00e9rrez D, Ferreras MC, P\u00e9rez V, Espinosa J (2020) Local assessment of WC1+ \u03b3\u03b4 T lymphocyte subset in the different types of lesions associated with bovine paratuberculosis. Comp Immunol Microbiol Infect Dis 69:101422. https:\/\/doi.org\/10.1016\/j.cimid.2020.101422","journal-title":"Comp Immunol Microbiol Infect Dis"},{"key":"925_CR22","doi-asserted-by":"publisher","first-page":"156","DOI":"10.1128\/CVI.00496-12","volume":"20","author":"C Charavaryamath","year":"2013","unstructured":"Charavaryamath C, Gonzalez-Cano P, Fries P, Gomis S, Doing K, Scruten E, Potter A, Napper S, Griebel PJ (2013) Host responses to persistent Mycobacterium avium subspecies paratuberculosis infection in surgically isolated bovine ileal segments. Clin Vaccine Immunol 20:156\u2013165. https:\/\/doi.org\/10.1128\/CVI.00496-12","journal-title":"Clin Vaccine Immunol"},{"key":"925_CR23","doi-asserted-by":"publisher","first-page":"275","DOI":"10.1016\/0165-2427(96)05578-x","volume":"52","author":"WC Davis","year":"1996","unstructured":"Davis WC, Brown WC, Hamilton MJ, Wyatt CR, Orden JA, Khalid AM, Naessens J (1996) Analysis of monoclonal antibodies specific for the gamma delta TcR. Vet Immunol Immunopathol 52:275\u2013283. https:\/\/doi.org\/10.1016\/0165-2427(96)05578-x","journal-title":"Vet Immunol Immunopathol"},{"key":"925_CR24","doi-asserted-by":"publisher","first-page":"340","DOI":"10.1046\/j.1365-2567.1997.00350.x","volume":"92","author":"ND Machugh","year":"1997","unstructured":"Machugh ND, Mburu JK, Carol MJ, Wyatt CR, Orden JA, Davis WC (1997) Identification of two distinct subsets of bovine gamma delta T cells with unique cell surface phenotype and tissue distribution. Immunology 92:340\u2013345. https:\/\/doi.org\/10.1046\/j.1365-2567.1997.00350.x","journal-title":"Immunology"},{"key":"925_CR25","doi-asserted-by":"publisher","first-page":"2089","DOI":"10.1128\/IAI.00779-10","volume":"79","author":"M Robinson","year":"2011","unstructured":"Robinson M, O\u2019Brien R, Mackintosh C, Clark R, Griffin J (2011) Immunoregulatory cytokines are associated with protection from immunopathology following Mycobacterium avium subspecies paratuberculosis infection in red deer. Infect Immun 79:2089\u20132097","journal-title":"Infect Immun"},{"key":"925_CR26","doi-asserted-by":"publisher","first-page":"137","DOI":"10.1038\/labinvest.3700189","volume":"85","author":"M Oki","year":"2005","unstructured":"Oki M, Ohtani H, Kinouchi Y, Sato E, Nakamura S, Matsumoto T, Nagura H, Yoshie O, Shimosegawa T (2005) Accumulation of CCR5+ T cells around RANTES+ granulomas in Crohn\u2019s disease: a pivotal site of Th1-shifted immune response? Lab Invest 85:137\u2013145","journal-title":"Lab Invest"},{"key":"925_CR27","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1186\/1471-2172-9-25","volume":"9","author":"U Singh","year":"2008","unstructured":"Singh U, Singh R, Singh S, Karls R, Quinn F, Taub D, Lilliard J Jr (2008) CXCL10+ T cells and NK cells assist in the recruitment and activation of CXCR3+ and CXCL11+ leukocytes during Mycobacteria-enhanced colitis. BMC Immunol 9:25","journal-title":"BMC Immunol"},{"key":"925_CR28","doi-asserted-by":"publisher","first-page":"46695","DOI":"10.1038\/srep46695","volume":"7","author":"A Gossner","year":"2017","unstructured":"Gossner A, Watkins C, Chianini F, Hopkins J (2017) Pathways and genes associated with immune dysfunction in sheep paratuberculosis. Sci Rep 7:46695. https:\/\/doi.org\/10.1038\/srep46695","journal-title":"Sci Rep"},{"key":"925_CR29","doi-asserted-by":"publisher","first-page":"974","DOI":"10.1128\/CVI.00324-15","volume":"22","author":"WJ Goosen","year":"2015","unstructured":"Goosen WJ, Cooper D, Miller MA, van Helden PD, Parsons SDC (2015) IP-10 is a sensitive biomarker of antigen recognition in whole-blood stimulation assays used for the diagnosis of Mycobacterium bovis infection in african buffaloes (Syncerus caffer). Clin Vaccine Immunol 22:974\u2013978. https:\/\/doi.org\/10.1128\/CVI.00324-15","journal-title":"Clin Vaccine Immunol"},{"key":"925_CR30","doi-asserted-by":"publisher","first-page":"198","DOI":"10.1016\/j.vetimm.2014.10.008","volume":"162","author":"WJ Goosen","year":"2014","unstructured":"Goosen WJ, Cooper D, Warren RM, Miller MA, van Helden PD, Parsons SDC (2014) The evaluation of candidate biomarkers of cell-mediated immunity for the diagnosis of Mycobacterium bovis infection in African buffaloes (Syncerus caffer). Vet Immunol Immunopathol 162:198\u2013202. https:\/\/doi.org\/10.1016\/j.vetimm.2014.10.008","journal-title":"Vet Immunol Immunopathol"}],"container-title":["Veterinary Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13567-021-00925-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s13567-021-00925-x\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13567-021-00925-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,4,13]],"date-time":"2021-04-13T09:13:19Z","timestamp":1618305199000},"score":1,"resource":{"primary":{"URL":"https:\/\/veterinaryresearch.biomedcentral.com\/articles\/10.1186\/s13567-021-00925-x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,13]]},"references-count":30,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2021,12]]}},"alternative-id":["925"],"URL":"https:\/\/doi.org\/10.1186\/s13567-021-00925-x","relation":{},"ISSN":["1297-9716"],"issn-type":[{"value":"1297-9716","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,13]]},"assertion":[{"value":"25 August 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 March 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"13 April 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"All animals were housed in American Association for Accreditation of Laboratory Animal Care-accredited facilities, and all animal related procedures were approved by the IACUC (National Animal Disease Center, Ames, IA) under protocol#ARS-722.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"The authors declare that they have no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"55"}}