The demographic data of study groups are presented in Table 1. The study was approved by the Ethics Committee of the Medical University of Warsaw.
The venous blood samples were collected before breakfast, early morning. All the analysis Selleckchem Paclitaxel were performed right after blood collection. First, anti-CD45-FITC and anti-CD14-PE was used for the lymphocyte gate setting at FSC/SSC graph. As a negative isotype controls the Ig2a-FITC and Ig2b-PE were applied. We analysed the proportion of following lymphocyte subtypes: T cells, B cells, T helper and T cytotoxic cells and the expression of CD25 and CTLA4 on CD4+ cells and CD25 on CD8+ cells with following mixtures of antibodies: CD3-FITC/CD19-PE (Becton-Dickinson Immunocytometry Systems, San Jose, CA, USA), CD4-FITC/CD8-PE, CD4-FITC/CD25-PE/CTLA4-Cy5, CD8-FITC/CD25-PE (Dako Cytomation, Glostrup, Denmark). The analyses were performed using three-colour flow cytometry method (FACS Calibur flow cytometer, Becton-Dickinson, San Jose, CA, USA). The cells were collected by Cellquest software. The analysis was performed in the same manner, with the same set of antibody and in the same conditions in patients and controls. The population of CD25high cells was gated manually and was well separated from
those with low CD25 expression (Fig. 1). The serum concentration of adiponectin PLX4032 mw was measured using Human Adiponectin/Acrp30 Immunoassay kit (R&D System, Minneapolis, MN, USA) and ELISA method according to the prescription by the producer. Statistical analysis. For data Rutecarpine comparison the Mann–Whitney U-test was used, P < 0.05 regarded as significant. The relationships between the data were examined by the Spearman rank correlation coefficient. Correlations with both R ≥ 0.4 and P < 0.05 were considered relevant. To present the data we used proportion of cells. The absolute number of cells depends on the number of gated events
and on absolute number of lymphocytes. To present the proportion is more common in the literature and seems to be more objective in the comparative studies. In the analysis of the main lymphocyte subpopulations we found that the proportion of T cells was significantly higher in patients than in controls, so was the proportion of T cytotoxic cells (Table 2). The Th/Tc ratio was significantly lower in patients than in healthy subjects (1.12 versus 2.0, P = 0.03). The proportion of all CD4+/CD25+ cells and the population with high expression of CD25 on CD4+ cells defined as CD25high cell were shown in the Table 2, and on Fig. 2. The proportion of CD4+/CD25+ of all lymphocytes was significantly lower in COPD patients when compared with controls (median value was 15.3 versus 17.9%, respectively, P = 0.03). The proportion of CD25high cells in the COPD group was significantly lower than in controls, median value: 0.79% versus 1.54%, P = 0.027.