Gingival cell IL-2 and IL-4 in early-onset periodontitis

S. S. Manhart, R. A. Reinhardt, J. B. Payne, G. J. Seymour, E. Gemmell, J. K. Dyer, T. M. Petro

Research output: Contribution to journalArticle

61 Scopus citations

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 adsorption 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). Analysis of IL-2/IL-4 positive GTMC ratios revealed significant differences between EOP and GC for non-stimulated cultures (0.5 ± 0.2 vs. 2.9 ± 1.0, P < 0.02). Digital analysis indicated significantly greater areas of IL-4 staining for EOP than GC GTMC in non-stimulated cultures (P < 0.006). These data would support the hypothesis that altered IL-2/IL-4 activities are associated with the periodontitis lesion.

Original languageEnglish (US)
Pages (from-to)807-813
Number of pages7
JournalJournal of periodontology
Volume65
Issue number9
DOIs
StatePublished - 1994

Keywords

  • Porphyromonas gingivalis
  • cytokines
  • early onset/etiology
  • interleukin-4
  • periodontitis

ASJC Scopus subject areas

  • Periodontics

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