Nuntana AroonrerkSathit PichyangkulKosol YongvanitchitMahisorn WisetchangNoppadol Sa-Ard-lamStitaya SirisinhaRangsini MahanondaMahidol UniversityArmed Forces Research Institute of Medical Sciences, ThailandFaculty of Medicine, Siriraj Hospital, Mahidol UniversityChulalongkorn University2018-07-242018-07-242003-01-01Journal of Periodontal Research. Vol.38, No.3 (2003), 262-268002234842-s2.0-0043261610https://repository.li.mahidol.ac.th/handle/20.500.14594/20841Objectives and background: It is well documented that in periodontitis lesions, most infiltrated gingival T cells are antigen-specific memory T cells. These cells play an important role as regulators and effector cells in the pathogenesis of periodontitis. In this study, we used dendritic cells (DCs) as antigen-presenting cells to generate human gingival T cell lines and clones specific for Porphyromonas gingivalis from periodontitis patients. Methods: Autologous DCs were derived from the patients' adherent monocytes using granulocyte-macrophage colony-stimulating factor and interleukin (IL)-4. Lymphocytes were isolated from gingival biopsies using collagenase enzyme digestion and the number was increased by subsequent culturing in IL-2-containing medium. T cells were then negatively sorted using flow cytometry, cocultured with P. gingivalis-pulsed DCs and subsequently expanded in the culture medium containing IL-2. T cells were kept viable and active by periodic exposure to antigen-pulsed DCs. The specificity of the T cell lines was tested against four plaque bacteria: P. gingivalis, Actinobacillus actinomycetemcomitans, Prevotella intermedia and Actinomyces viscosus. The established T cell lines were then cloned. Three P. gingivalis-specific T cell lines and 12 gingival T cell clones were generated. They all showed good specificity against P. gingivalis but not to other plaque bacteria. Results: All T cell clones were positive for CD4 and the majority of them produced interferon gamma, but a minimal or negligible amount of IL-5. Conclusions: The data obtained clearly showed that monocyte-derived DCs could be used as powerful antigen-presenting cells to generate antigen-specific T cells from periodontitis tissues.Mahidol UniversityDentistryArticleSCOPUS10.1034/j.1600-0765.2003.02658.x