title: Ex vivo CD(+) T-cell cytokine expression from patients with Sjögren's syndrome following in vitro stimulation to induce proliferation.
abstract: OBJECTIVE
  To assess ex vivo CD4(+) T-cell cytokine expression from patients with primary Sjögren's syndrome (SS) following in vitro stimulation to induce proliferation, as proliferation is closely related to differentiation of cytokine-producing cells.
  
  
  METHODS
  Peripheral blood mononuclear cells (PBMCs) separated from primary SS patients (n = 28) and controls (n = 25) were analysed. PBMCs were stimulated with concanavalin A followed by phorbol 12-myristate 13-acetate and ionomycin. Intracellular interferon-gamma (IFN-gamma) and interleukin-4 (IL)-4 in proliferating CD4(+) T cells were assessed by flow cytometry. The proportion of cytokine-producing cells and proliferating cells in each division cycle was assessed using [5(and 6)-carboxyfluorescein diacetate, succinimidyl ester]-labelled CD4(+/-) T cells.
  
  
  RESULTS
  The proportion of IFN-gamma+ proliferating CD4(+) T cells in each cell division cycle from extraglandular SS was increased in glandular SS patients compared glandular SS patients with controls (P<0.05 approximately 0.01). The percentage of IFN-gamma single positive proliferating CD4(+) T cells was greater in extraglandular SS patients (26.7+/-14.1%) compared with glandular SS (9.9 +/- 9.1%) (P<0.01) and controls (9.4 +/- 5.8%) (P<0.001). There was no significant difference in the percentages of IL-4(+) proliferating CD4(+) T cells among the groups. However, the proliferating response of CD4(+) T cells was significantly decreased in extraglandular SS patients (percentage of proliferating cells 38.4 +/- 18.6%) compared with that in glandular SS patients (64.2 +/- 17.2%) (P<0.05) and controls (63.1+/-10.6%) (P<0.01).
  
  
  CONCLUSIONS
  CD4(+) T cells from extraglandular SS patients may have a predisposition for entry into the IFN-gamma-producing effector pathway as a result of the stimulations. These results are helpful for understanding the immunological difference between glandular and extraglandular SS and the mechanisms of disease progression.
authors.affiliation: Division of Rheumatology, Saga Medical School, 5-1-1 Nabeshima, Saga 849-8501, Japan. koarada@med.saga-u.ac.jp
authors.forename: S
authors.initials: S
authors.lastname: Koarada
authors.forename: Y
authors.initials: Y
authors.lastname: Haruta
authors.forename: M
authors.initials: M
authors.lastname: Mitamura
authors.forename: F
authors.initials: F
authors.lastname: Morito
authors.forename: Y
authors.initials: Y
authors.lastname: Tada
authors.forename: A
authors.initials: A
authors.lastname: Ohta
authors.forename: K
authors.initials: K
authors.lastname: Nagasawa
mesh_terms: D000328:Adult; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002455:Cell Division / Q000276:immunology; D002478:Cells, Cultured; D003208:Concanavalin A / Q000276:immunology; D016207:Cytokines / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology; D015847:Interleukin-4 / Q000276:immunology; D015759:Ionomycin / Q000276:immunology; D007476:Ionophores / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008297:Male; D008875:Middle Aged; D008934:Mitogens / Q000276:immunology; D012859:Sjogren's Syndrome / Q000276:immunology*; D013755:Tetradecanoylphorbol Acetate / Q000276:immunology
chemical_list: D016207:Cytokines; D007476:Ionophores; D008934:Mitogens; D003208:Concanavalin A; D015847:Interleukin-4; D015759:Ionomycin; D007371:Interferon-gamma; D013755:Tetradecanoylphorbol Acetate