Generation of gingival T cell lines/clones specific with Porphyromonas gingivalis pulsed dendritic cells from periodontitis patients.
OBJECTIVE
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.
Aroonrerk
Nuntana
N
Department of Microbiology, Faculty of Science, Mahidol University, Bangkok, Thailand.
Pichyangkul
Sathit
S
Yongvanitchit
Kosol
K
Wisetchang
Mahisorn
M
Sa-Ard-Iam
Noppadol
N
Sirisinha
Stitaya
S
Mahanonda
Rangsini
R
D016924:Actinomyces viscosus / Q000276:immunology; D000328:Adult; D016976:Aggregatibacter actinomycetemcomitans / Q000276:immunology; D000938:Antigen-Presenting Cells / Q000276:immunology; D000942:Antigens, Bacterial / Q000276:immunology; D015704:CD4 Antigens / Q000276:immunology; D002460:Cell Line; D002999:Clone Cells; D003713:Dendritic Cells / Q000276:immunology*; D003773:Dental Plaque / Q000382:microbiology; D000939:Epitopes / Q000276:immunology; D005881:Gingiva / Q000276:immunology*; D006801:Humans; D007156:Immunologic Memory / Q000276:immunology; D007371:Interferon-gamma / Q000276:immunology; D015848:Interleukin-5 / Q000276:immunology; D008875:Middle Aged; D009000:Monocytes / Q000276:immunology; D010518:Periodontitis / Q000276:immunology*; D016966:Porphyromonas gingivalis / Q000276:immunology*; D018713:Prevotella intermedia / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*
D000942:Antigens, Bacterial; D015704:CD4 Antigens; D000939:Epitopes; D015848:Interleukin-5; D007371:Interferon-gamma