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Category Archives: Profile of interleukin-10
Different profile of interleukin-10 production: DISCUSSION (3)
Stimulation with PMA and ionomycin increased the frequencies of IL-10-producing CD4+ and CD8+ T cells in healthy controls and modestly in atopic nonasthmatics, but not in asthmatics. These results were surprising because we had expected that additional stimulus would enhance T cell IL-10 production in asthmatics, in whom the frequency of IL-10-producing T cells was increased at baseline. (more…)
Different profile of interleukin-10 production: DISCUSSION (2)
At unstimulated baseline, the frequencies of IL-10-producing CD4+ and CD8+ T cells were significantly higher in atopic asthmatics than in healthy controls. A similar difference was noted in CD4+ T cells between atopic nonasthmatics and healthy controls. T cells circulate systemically from the affected organs to the blood stream via the lymphatic system . Our findings in the peripheral blood may reflect the increased numbers of IL-10-producing T cells in the airways of atopic asthmatics. (more…)
Different profile of interleukin-10 production: DISCUSSION (1)
This study has demonstrated that the frequencies of IL-10-producing CD4+ and CD8+ T cells were higher in atopic asthmatics than in healthy controls. In atopic nonasthmatics, the frequency of IL-10-producing CD4+, but not CD8+, T cells was higher than in healthy controls. Furthermore, stimulation with PMA and ionomycin increased the frequencies of IL-10-producing CD4+ and CD8+ T cells in healthy controls and atopic nonasthmatics, but not in asthmatics. (more…)
Different profile of interleukin-10 production: RESULTS (2)
Frequency of IL-10-producing T cells at baseline
To evaluate the spontaneous production of IL-10, the frequencies of IL-10-producing cells, cultured for 4h without stimulation, were compared among groups. The frequency of IL-10-producing CD3+ cells in asthmatics was significantly higher than in healthy controls (P=0.008) (Figure 2, Top). The values from atopic nonasthmatics showed an intermediate distribution between healthy controls and asthmatics. (more…)
Different profile of interleukin-10 production: RESULTS (1)
PBMCs from six healthy controls were examined for the relative frequencies of IL-10-producing cells after stimulation with PMA and ionomycin (Table 1). The frequencies of IL-10-producing CD3+ cells after 4 h, 8 h and 24 h of stimulation were all significantly higher than that of the control samples (P<0.05). A typical dot plot is shown in Figure 1. Similarly, the frequencies of IL-10-producing CD4+ cells after 4 h, 8 h and 24 h of stimulation were significantly higher than that of the control samples (P<0.05). (more…)
Different profile of interleukin-10 production: PATIENTS AND METHODS (5)
An analysis gate was set on the lymphocyte population based on the forward and side scatter plot. Firstly, CD3+-stained cells falling within the gated area were identified. The gated population was further analyzed for the identification of CD3+CD4+ (CD4+) or CD3+CD8+ (CD8+). Previous studies have shown that the stimulation with PMA and calcium ionophore downregulates surface CD4 expression on T cells, which may make it difficult to identify CD4+ T cells . (more…)
Different profile of interleukin-10 production: PATIENTS AND METHODS (3)
Cell cultures
PBMCs were isolated by Lymphoprep (Nycomed, Norway) density gradient separation and washed with RPMI-1640 (Gibco BRL, USA). Cells were suspended in RPMI-1640 with 10% fetal calf serum (Gibco BRL, USA) and 25 mM of Hepes (Gibco BRL) at a density of 2x106 cells/mL. Cells were stimulated in six-well culture plates (Becton Dickinson Labware, USA) in a volume of 5 mL with phorbol 12-myris-tate 13-acetate (PMA) (Sigma, USA) at a concentration of 10 ng/mL plus ionomycin (Sigma, USA) at a concentration of 2 pM for 4 h, 8 h and 24 h at 37°C in 5% carbon dioxide in an incubator. (more…)
Different profile of interleukin-10 production: PATIENTS AND METHODS (2)
The healthy control group consisted of six male and six female subjects (median age 35 years, range 27 to 49 years) who had no history of allergic diseases and negative skin tests. All subjects were nonsmokers, were not exposed to allergens and had not had exacerbations of symptoms or respiratory tract infections for at least two weeks before the investigation. (more…)
Different profile of interleukin-10 production: PATIENTS AND METHODS (1)
Patients
Thirty-five subjects were studied, of whom 12 were nonatopic, healthy subjects, 10 were atopic nonasthmatics and 13 were atopic asthmatics. All patients gave signed consent before the study. The study was approved by the Ethics Committee of the McMaster University Health Sciences Centre, Hamilton, Ontario. The asthmatic group consisted of nine male and four female patients with a median age of 35 years (range 22 to 50 years). (more…)
Different profile of interleukin-10 production in circulating T cells from atopic asthmatics compared with healthy subjects (2)
Several studies reported that IL-10 levels in bronchoalveolar lavage (BAL) fluid and in the supernatant of cultured alveolar macrophages were lower in asthmatics than in healthy subjects . However, other studies have shown that levels of IL-10 messenger ribonucleic acid (mRNA) in airway tissues and its protein in BAL fluid were higher in asthmatics . A decreased level of IL-10 protein in the supernatant of peripheral blood mononuclear cells (PBMCs) from asthmatics was reported by some investigators , but not by others . (more…)