sq-23377 and Rhinitis--Allergic--Seasonal

Human immune cells, included T helper cells and T cytotoxic/suppressor cells, may be divided into two distinct subtypes according to the profile of secreted cytokines. The allergic disorders are characterised by type 2 lymphocyte response, while the autoimmune diseases are associated with type 1 response. The aim of this study was to assess the usefulness of flow cytometric analysis of intracellular expression of cytokines typical for type 2 (IL-4) and type 1 response (IFN-gamma) in helper and cytotoxic/suppressor T cells in patients with chronic urticaria or intermittent allergic rhinitis. 10 subjects (6 male) with intermittent allergic rhinitis (a group ALER) and 6 subjects (1 male) with chronic urticaria (a group PP) were included into the study. The intracellular expression of IL-4 and IFN-gamma in CD4+ or CD8+ cells after stimulation with ionomycin/PMA was estimated by flow cytometer (FACSCalibur, Becton Dickinson) and serum levels of both cytokines were assessed with ELISA (R & D Systems) in all subjects. The percentage of CD4+ cell producing IFN-gamma was statistically higher in the PP group than in the ALER group, and the percentage of IL-4 producing CD8+ cells was also higher, although non significantly. The intracellular expression of IL-4 was comparable in both cell populations in the two examined groups. The surface expression of CD4 diminished after stimulation with PMA in all subjects. No correlations between serum level and intracellular expression of the examined cytokines were observed.. A tendency to the predominance of type 1 response of T lymphocytes was observed in chronic urticaria compared to intermittent allergic rhinitis.

Topics: Adult; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Humans; Interferon-gamma; Interleukin-4; Ionomycin; Male; Middle Aged; Rhinitis, Allergic, Seasonal; Th1 Cells; Th2 Cells; Urticaria

As a plasma membrane pore-forming protein, perforin is essential for T-cell cytotoxicity mediated by lytic granules. Recent studies on the immune system of perforin knockout mice demonstrated striking similarities to the immunopathology of atopic diseases.. We sought to investigate the perforin system of atopic patients.. Monoclonal antibodies were used to characterize perforin-positive PBMCs of patients with exacerbated atopic dermatitis (AD) and asymptomatic rhinoconjunctivitis allergica (RCA) by means of immunoflow cytometry. In addition, a perforin release assay was developed to quantify the velocity of ionomycin and phorbol 12-myristate 13-acetate-induced secretion of lytic granules.. In atopic patients significantly fewer lymphocytes contained perforin-positive lytic granules compared with those of healthy control subjects (patients with AD: 14% +/- 5%, n = 13, P <.0001; patients with RCA: 24% +/- 5%, n = 9, P <.01; healthy control subjects: 33% +/- 11%, n = 13). Of all CD8(hi+) cytotoxic T lymphocytes (CTLs), only 18% +/- 9% and 17% +/- 12% were perforin-positive in patients with AD and RCA, respectively, compared with 44% +/- 13% in control subjects (P <.0001). In addition, perforin-positive CD8(hi+) CTLs of atopic patients released their perforin twice as fast and more completely than control CTLs. This means that 50% of initially perforin-positive CD8(hi+) CTLs from patients with AD and RCA released their perforin completely within 32 +/- 16 and 36 +/- 19 minutes, respectively, and an over 85% release was reached within 113 +/- 41 and 118 +/- 60 minutes, respectively. In CTLs of healthy control subjects, however, it took 64 +/- 40 minutes to achieve a 50% release of lytic granules, and an 85% depletion was not reached in 60% of healthy control subjects, even after 180 minutes.. The perforin hyperreleasability explains, at least in part, the decreased percentage of perforin-positive CD8(hi+) CTLs in atopic patients. These distortions in the system of lytic granules of atopic patients may contribute to the functional defects observed in T-cell cytotoxicity in vivo and in vitro in patients with AD and RCA.

Topics: Antibodies, Monoclonal; Antigens, Differentiation, T-Lymphocyte; Antigens, Neoplasm; CD4-CD8 Ratio; Conjunctivitis, Allergic; Cytoplasmic Granules; Cytotoxicity, Immunologic; Dermatitis, Atopic; Flow Cytometry; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Ionomycin; Killer Cells, Natural; Membrane Glycoproteins; Perforin; Pore Forming Cytotoxic Proteins; Rhinitis, Allergic, Perennial; Rhinitis, Allergic, Seasonal; T-Lymphocytes, Cytotoxic; Tetradecanoylphorbol Acetate; Th1 Cells; Th2 Cells; Time Factors