{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T03:14:15Z","timestamp":1789528455688,"version":"build-2803163510"},"reference-count":46,"publisher":"Oxford University Press (OUP)","issue":"3","license":[{"start":{"date-parts":[[2012,5,30]],"date-time":"2012-05-30T00:00:00Z","timestamp":1338336000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"name":"Hack Memorial Foundation, University of Washington"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2012,5,30]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>A novel oral mucosal gingival explant culture facilitates isolation of tissue resident mononuclear cells that represent tissue resident population, and are functional.<\/jats:p>\n               <jats:p>During CP, the gingival environment is primed to recruit and activate homing lymphocytes. However, detailed phenotypic and functional characterization of gingival tissue resident lymphocytes has been challenging as a result of limitations associated with available isolation methods and limited availability of human samples. This study aimed to develop a novel explant culture technique for effectively isolating human gingival lymphocytes. This technique takes advantage of the natural tendency of MNCs to migrate toward a chemokine gradient generated by the gingival fibroblasts. The explant system allowed isolation of MNCs with \u223c95\u00d7 higher yield relative to conventional approaches. The MNC yield correlates directly with wet weights of the tissues, and maximal MNCs are isolated during the 2nd day of the culture. The explant culture method and conventional approach produce similar MNC subpopulations such as Th, Tc, and B cells. Chemokines associated with MNC migration but not cytokines associated with MNC proliferation and differentiation were produced in the explant culture. Moreover, MNC migration in response to the secreted chemokines was inhibited by PTX. T cells did not undergo proliferation during the culture. However, the isolated T cells responded to mitogenic stimulation ex vivo. A statistically, significantly decreased Th cell with reduced CD25 expression along with increased NK and NKT cells in CP are shown. However, the number of na\u00efve NK cells was decreased significantly in MNCs, suggesting activation of NK cells in CP.<\/jats:p>","DOI":"10.1189\/jlb.0212103","type":"journal-article","created":{"date-parts":[[2012,5,31]],"date-time":"2012-05-31T05:55:49Z","timestamp":1338443749000},"page":"683-692","source":"Crossref","is-referenced-by-count":14,"title":["Technical Advance: Decreased helper T cells and increased natural killer cells in chronic periodontitis analyzed by a novel method for isolating resident lymphocytes"],"prefix":"10.1093","volume":"92","author":[{"given":"Manoj","family":"Muthukuru","sequence":"first","affiliation":[{"name":"University of Washington, Department of Periodontics, School of Dentistry , Seattle, Washington, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2012,5,30]]},"reference":[{"key":"2023012312545054300_B1","doi-asserted-by":"crossref","first-page":"3770","DOI":"10.1128\/JB.183.12.3770-3783.2001","article-title":"Bacterial diversity in human subgingival plaque","volume":"183","author":"Paster","year":"2001","journal-title":"J. Bacteriol"},{"key":"2023012312545054300_B2","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.imlet.2004.02.005","article-title":"Commensal bacteria (normal microflora), mucosal immunity and chronic inflammatory and autoimmune diseases","volume":"93","author":"Tlaskalova-Hogenova","year":"2004","journal-title":"Immunol Lett."},{"key":"2023012312545054300_B3","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1046\/j.1462-5822.2002.00192.x","article-title":"Involvement of integrins in fimbriae-mediated binding and invasion by","volume":"4","author":"Yilmaz","year":"2002","journal-title":"Porphyromonas gingivalis. Cell. Microbiol"},{"key":"2023012312545054300_B4","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1111\/j.1600-051X.1991.tb02310.x","article-title":"Importance of the host response in the periodontium","volume":"8","author":"Seymour","year":"1991","journal-title":"J. Clin. Periodontol. 1"},{"key":"2023012312545054300_B5","doi-asserted-by":"crossref","first-page":"14547","DOI":"10.1073\/pnas.96.25.14547","article-title":"Bacterial diversity within the human subgingival crevice","volume":"96","author":"Kroes","year":"1999","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2023012312545054300_B6","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1046\/j.0906-6713.2003.03203.x","article-title":"Microorganisms as risk indicators for periodontal disease","volume":"32","author":"Ezzo","year":"2003","journal-title":"Periodontol. 2000"},{"key":"2023012312545054300_B7","doi-asserted-by":"crossref","first-page":"736","DOI":"10.1177\/154405910308200915","article-title":"Multiple dendritic cell (DC) subpopulations in human gingiva and association of mature DCs with CD4+ T-cells in situ","volume":"82","author":"Jotwani","year":"2003","journal-title":"J. Dent. Res."},{"key":"2023012312545054300_B8","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1902\/jop.2002.73.1.45","article-title":"Lamina propria dendritic cells express activation markers and contact lymphocytes in chronic periodontitis","volume":"73","author":"Cirrincione","year":"2002","journal-title":"J. Periodontol."},{"key":"2023012312545054300_B9","doi-asserted-by":"crossref","first-page":"1263","DOI":"10.1016\/S0002-9440(10)64077-1","article-title":"Macrophage-derived chemokine (MDC\/CCL22) and CCR4 are involved in the formation of T lymphocyte-dendritic cell clusters in human inflamed skin and secondary lymphoid tissue","volume":"158","author":"Katou","year":"2001","journal-title":"Am. J. Pathol."},{"key":"2023012312545054300_B10","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1016\/S0002-9440(10)64756-6","article-title":"Immunological activation of dermal Langerhans cells in contact with lymphocytes in a model of human inflamed skin","volume":"156","author":"Katou","year":"2000","journal-title":"Am. J. Pathol."},{"key":"2023012312545054300_B11","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1111\/j.1600-0714.1992.tb00961.x","article-title":"Distribution and phenotype of immune cells in normal human gingiva: active immune response versus unresponsiveness","volume":"21","author":"Colasante","year":"1992","journal-title":"J. Oral Pathol Med."},{"key":"2023012312545054300_B12","doi-asserted-by":"crossref","first-page":"4693","DOI":"10.4049\/jimmunol.167.8.4693","article-title":"Mature dendritic cells infiltrate the T cell-rich region of oral mucosa in chronic periodontitis: in situ, in vivo, and in vitro studies","volume":"167","author":"Jotwani","year":"2001","journal-title":"J. Immunol."},{"key":"2023012312545054300_B13","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1177\/154405910608500301","article-title":"Protective and destructive immunity in the periodontium: Part 1\u2014innate and humoral immunity and the periodontium","volume":"85","author":"Teng","year":"2006","journal-title":"J. Dent. Res."},{"key":"2023012312545054300_B14","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1177\/154405910608500302","article-title":"Protective and destructive immunity in the periodontium: Part 2\u2014T-cell-mediated immunity in the periodontium","volume":"85","author":"Teng","year":"2006","journal-title":"J. Dent. Res."},{"key":"2023012312545054300_B15","doi-asserted-by":"crossref","first-page":"656","DOI":"10.2741\/1560","article-title":"Dendritic cells in rheumatoid arthritis","volume":"10","author":"Sarkar","year":"2005","journal-title":"Front. Biosci."},{"key":"2023012312545054300_B16","doi-asserted-by":"crossref","first-page":"877","DOI":"10.4049\/jimmunol.166.2.877","article-title":"CCR7 expression and memory T cell diversity in humans","volume":"166","author":"Campbell","year":"2001","journal-title":"J. Immunol."},{"key":"2023012312545054300_B17","doi-asserted-by":"crossref","first-page":"1265","DOI":"10.1084\/jem.20032177","article-title":"A skin-selective homing mechanism for human immune surveillance T cells","volume":"199","author":"Schaerli","year":"2004","journal-title":"J. Exp. Med."},{"key":"2023012312545054300_B18","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1006\/cyto.2001.0836","article-title":"Cytokine-augmented culture of haematopoietic progenitor cells in a novel three-dimensional cell growth matrix","volume":"13","author":"Banu","year":"2001","journal-title":"Cytokine"},{"key":"2023012312545054300_B19","doi-asserted-by":"crossref","first-page":"1431","DOI":"10.1128\/IAI.74.2.1431-1435.2006","article-title":"Upregulation of immunoregulatory Src homology 2 molecule containing inositol phosphatase and mononuclear cell hyporesponsiveness in oral mucosa during chronic periodontitis","volume":"74","author":"Muthukuru","year":"2006","journal-title":"Infect. Immun."},{"key":"2023012312545054300_B20","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1128\/IAI.00100-07","article-title":"Antigen capture of Porphyromonas gingivalis by human macrophages is enhanced but killing and antigen presentation are reduced by endotoxin tolerance","volume":"76","author":"Muthukuru","year":"2008","journal-title":"Infect. Immun."},{"key":"2023012312545054300_B21","doi-asserted-by":"crossref","first-page":"687","DOI":"10.1128\/IAI.73.2.687-694.2005","article-title":"Oral mucosal endotoxin tolerance induction in chronic periodontitis","volume":"73","author":"Muthukuru","year":"2005","journal-title":"Infect. Immun."},{"key":"2023012312545054300_B22","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1002\/jlb.57.2.335","article-title":"Modulation of IL-8 receptor expression on purified human T lymphocytes is associated with changed chemotactic responses to IL-8","volume":"57","author":"Xu","year":"1995","journal-title":"J. Leukoc. Biol."},{"key":"2023012312545054300_B23","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1002\/(SICI)1521-4141(199801)28:01<104::AID-IMMU104>3.0.CO;2-F","article-title":"Transendothelial chemotaxis of human \u03b1\/\u03b2 and \u03b3\/\u03b4 T lymphocytes to chemokines","volume":"28","author":"Roth","year":"1998","journal-title":"Eur. J. Immunol."},{"key":"2023012312545054300_B24","doi-asserted-by":"crossref","first-page":"776","DOI":"10.1038\/23495","article-title":"The chemokine receptor CCR4 in vascular recognition by cutaneous but not intestinal memory T cells","volume":"400","author":"Campbell","year":"1999","journal-title":"Nature"},{"key":"2023012312545054300_B25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1101\/SQB.1989.054.01.003","article-title":"Approaching the asymptote? Evolution and revolution in immunology","volume":"54","author":"Janeway","year":"1989","journal-title":"Cold Spring Harb. Symp. Quant. Biol."},{"key":"2023012312545054300_B26","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/0167-5699(92)90198-G","article-title":"The immune system evolved to discriminate infectious nonself from noninfectious self","volume":"13","author":"Janeway","year":"1992","journal-title":"Immunol. Today"},{"key":"2023012312545054300_B27","doi-asserted-by":"crossref","first-page":"2444","DOI":"10.4049\/jimmunol.166.4.2444","article-title":"Microbial lipopeptides stimulate dendritic cell maturation via Toll-like receptor 2","volume":"166","author":"Hertz","year":"2001","journal-title":"J. Immunol."},{"key":"2023012312545054300_B28","doi-asserted-by":"crossref","first-page":"2039","DOI":"10.4049\/jimmunol.172.4.2039","article-title":"Inhibitory effect of Toll-like receptor 4 on fusion between phagosomes and endosomes\/lysosomes in macrophages","volume":"172","author":"Shiratsuchi","year":"2004","journal-title":"J. Immunol."},{"key":"2023012312545054300_B29","doi-asserted-by":"crossref","first-page":"3804","DOI":"10.4049\/jimmunol.165.7.3804","article-title":"Activation of Toll-like receptor 2 on human dendritic cells triggers induction of IL-12, but not IL-10","volume":"165","author":"Thoma-Uszynski","year":"2000","journal-title":"J. Immunol."},{"key":"2023012312545054300_B30","doi-asserted-by":"crossref","first-page":"7016","DOI":"10.4049\/jimmunol.151.12.7016","article-title":"Inhibition of C3 and IgG proteolysis enhances phagocytosis of","volume":"151","author":"Cutler","year":"1993","journal-title":"Porphyromonas gingivalis. J. Immunol."},{"key":"2023012312545054300_B31","doi-asserted-by":"crossref","first-page":"1725","DOI":"10.1128\/IAI.72.3.1725-1732.2004","article-title":"Fimbriated Porphyromonas gingivalis is more efficient than fimbria-deficient P. gingivalis in entering human dendritic cells in vitro and induces an inflammatory Th1 effector response","volume":"72","author":"Jotwani","year":"2004","journal-title":"Infect. Immun."},{"key":"2023012312545054300_B32","doi-asserted-by":"crossref","first-page":"542","DOI":"10.1111\/j.1600-051X.1990.tb01103.x","article-title":"Immunohistological analysis of gingival lymphocytes in adult periodontitis","volume":"17","author":"Celenligil","year":"1990","journal-title":"J. Clin. Periodontol."},{"key":"2023012312545054300_B33","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1902\/jop.1987.58.8.569","article-title":"Phenotypic and functional analysis of T cells extracted from chronically inflamed human periodontal tissues","volume":"58","author":"Cole","year":"1987","journal-title":"J. Periodontol."},{"key":"2023012312545054300_B34","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1902\/jop.1993.64.2.120","article-title":"In situ characterization of gingival mononuclear cells in rapidly progressive periodontitis","volume":"64","author":"Celenligil","year":"1993","journal-title":"J. Periodontol."},{"key":"2023012312545054300_B35","first-page":"889","article-title":"T cell regulation of the immune response to infection in periodontal diseases","volume":"18","author":"Yamazaki","year":"2003","journal-title":"Histol. Histopathol."},{"key":"2023012312545054300_B36","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1034\/j.1600-051X.1999.260309.x","article-title":"Relative proportions of mononuclear cell types in periodontal lesions analyzed by immunohistochemistry","volume":"26","author":"Lappin","year":"1999","journal-title":"J. Clin. Periodontol."},{"key":"2023012312545054300_B37","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1177\/10454411010120020301","article-title":"Involvement of T-lymphocytes in periodontal disease and in direct and indirect induction of bone resorption","volume":"12","author":"Taubman","year":"2001","journal-title":"Crit. Rev. Oral Biol. Med."},{"key":"2023012312545054300_B38","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1080\/00016350152509247","article-title":"T-cell contributions to alveolar bone loss in response to oral infection with","volume":"59","author":"Baker","year":"2001","journal-title":"Porphyromonas gingivalis. Acta Odontol. Scand."},{"key":"2023012312545054300_B39","doi-asserted-by":"crossref","first-page":"R59","DOI":"10.1172\/JCI10763","article-title":"Functional human T-cell immunity and osteoprotegerin ligand control alveolar bone destruction in periodontal infection","volume":"106","author":"Teng","year":"2000","journal-title":"J. Clin. Invest."},{"key":"2023012312545054300_B40","doi-asserted-by":"crossref","first-page":"4652","DOI":"10.4049\/jimmunol.161.9.4652","article-title":"Stat3 activation is responsible for IL-6-dependent T cell proliferation through preventing apoptosis: generation and characterization of T cell-specific Stat3-deficient mice","volume":"161","author":"Takeda","year":"1998","journal-title":"J. Immunol."},{"key":"2023012312545054300_B41","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/S0065-2776(08)60509-X","article-title":"Interleukin-8 and related chemotactic cytokines\u2014CXC and CC chemokines","volume":"55","author":"Baggiolini","year":"1994","journal-title":"Adv. Immunol."},{"key":"2023012312545054300_B42","doi-asserted-by":"crossref","first-page":"1059","DOI":"10.1038\/sj.jid.5700199","article-title":"A novel method for the isolation of skin resident T cells from normal and diseased human skin","volume":"126","author":"Clark","year":"2006","journal-title":"J. Invest. Dermatol."},{"key":"2023012312545054300_B43","doi-asserted-by":"crossref","first-page":"1391","DOI":"10.1016\/S0002-9440(10)64090-4","article-title":"Elevated proportion of natural killer T cells in periodontitis lesions: a common feature of chronic inflammatory diseases","volume":"158","author":"Yamazaki","year":"2001","journal-title":"Am. J. Pathol."},{"key":"2023012312545054300_B44","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1111\/j.1600-0765.2005.00837.x","article-title":"Increased infiltration of CD1d and natural killer T cells in periodontal disease tissues","volume":"41","author":"Amanuma","year":"2006","journal-title":"J. Periodontal. Res."},{"key":"2023012312545054300_B45","doi-asserted-by":"crossref","first-page":"858","DOI":"10.1177\/154405910508400915","article-title":"Dendritic-NK cell interactions in P. gingivalis-specific responses","volume":"84","author":"Kikuchi","year":"2005","journal-title":"J. Dent. Res."},{"key":"2023012312545054300_B46","doi-asserted-by":"crossref","first-page":"5089","DOI":"10.1128\/IAI.72.9.5089-5096.2004","article-title":"Dendritic cells stimulated with Actinobacillus actinomycetemcomitans elicit rapid \u03b3 interferon responses by natural killer cells","volume":"72","author":"Kikuchi","year":"2004","journal-title":"Infect. Immun."}],"container-title":["Journal of Leukocyte Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/jleukbio\/article-pdf\/92\/3\/683\/48766391\/jlb0683.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jleukbio\/article-pdf\/92\/3\/683\/48766391\/jlb0683.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,2,27]],"date-time":"2023-02-27T13:13:59Z","timestamp":1677503639000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jleukbio\/article\/92\/3\/683\/6936116"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,5,30]]},"references-count":46,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2012,5,30]]},"published-print":{"date-parts":[[2012,5,30]]}},"URL":"https:\/\/doi.org\/10.1189\/jlb.0212103","relation":{},"ISSN":["1938-3673","0741-5400"],"issn-type":[{"value":"1938-3673","type":"electronic"},{"value":"0741-5400","type":"print"}],"subject":[],"published-other":{"date-parts":[[2012,9]]},"published":{"date-parts":[[2012,5,30]]}}}