title: Gingival cell IL-2 and IL-4 in early-onset periodontitis.
abstract: The purpose of this study was to compare, using cell blot analysis, the association of gingival tissue mononuclear cells (GTMC) isolated from lesions displaying histories of early-onset periodontitis (EOP; typically B-lymphocyte dominated) and gingivitis (typically T-lymphocyte dominated) with the B-cell stimulating cytokine, interleukin (IL)-4, and the T-cell stimulating cytokine, IL-2. Eleven EOP patients and 11 age- and gender-similar gingivitis control (GC) subjects participated. Gingival tissue adjacent to the alveolar crest normally removed during surgery was digested in collagenase-containing media and GTMC were isolated by density gradient centrifugation. Cells were separated into four aliquots. One was left unstimulated; the remainder were stimulated for 2 hours with Porphyromonas gingivalis outer membrane protein, mitogen Concanavalin A, or common antigen tetanus toxoid. Cells then were centrifuged onto transfer membranes and incubated in RPMI 1640 media for 6 hours to allow absorption of secreted cytokine. Membranes were treated with monoclonal anti-IL-2 or anti-IL-4, followed by a biotin-conjugated second layer, streptavidin-alkaline phosphatase and nitro blue tetrazolium/5-bromo-4-chloro-indolyl-phosphate (NBT/BCIP) color development. A higher percentage of GTMC from EOP patients were IL-2+ when stimulated with P. gingivalis compared with GTMC from GC patients (20 +/- 2% vs. 12 +/- 2%, P < 0.003). A higher percentage of non-stimulated GTMC from EOP patients produced IL-4 than from GC (22 +/- 4% vs. 6 +/- 3%, P < 0.00007), as well as when stimulated with P. gingivalis (22 +/- 3% vs. 13 +/- 2%, P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Surgical Specialties, University of Nebraska Medical Center, College of Dentistry, Lincoln.
authors.forename: S S
authors.initials: SS
authors.lastname: Manhart
authors.forename: R A
authors.initials: RA
authors.lastname: Reinhardt
authors.forename: J B
authors.initials: JB
authors.lastname: Payne
authors.forename: G J
authors.initials: GJ
authors.lastname: Seymour
authors.forename: E
authors.initials: E
authors.lastname: Gemmell
authors.forename: J K
authors.initials: JK
authors.lastname: Dyer
authors.forename: T M
authors.initials: TM
authors.lastname: Petro
mesh_terms: D000327:Adsorption; D000328:Adult; D010520:Aggressive Periodontitis / Q000276:immunology* / Q000473:pathology*; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology; D016022:Case-Control Studies; D002452:Cell Count; D002478:Cells, Cultured; D003208:Concanavalin A / Q000276:immunology; D003430:Cross-Sectional Studies; D005260:Female; D005881:Gingiva / Q000276:immunology* / Q000473:pathology*; D005891:Gingivitis / Q000276:immunology* / Q000473:pathology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007211:Indoles; D007376:Interleukin-2 / Q000032:analysis / Q000276:immunology*; D015847:Interleukin-4 / Q000032:analysis / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology; D008297:Male; D009580:Nitroblue Tetrazolium; D016966:Porphyromonas gingivalis / Q000276:immunology; D013745:Tetanus Toxoid / Q000276:immunology
chemical_list: D001425:Bacterial Outer Membrane Proteins; D007211:Indoles; D007376:Interleukin-2; D013745:Tetanus Toxoid; D003208:Concanavalin A; D015847:Interleukin-4; D009580:Nitroblue Tetrazolium; C035455:5-bromo-4-chloro-3-indoxyl phosphate