Fractions 2 to 6 were pooled (BAL fluid) and analyzed for differential cell count and the subsets and subpopulation of lymphocytes. Bronchoscopies and BAL were performed identically and by the same examiner (E.M.) in all patients and control subjects.

Cell Analysis in Canadian pharmacy

Differential cell count was performed as previously de-scribed. The frequency of T lymphocytes (CD3 + ), T-helper lymphocytes (CD4 + ), T-cytotoxic lymphocytes (CD8 + ), and activated T lymphocytes HLA-DR+ was assessed in BAL fluid and blood by dual-color flow cytometry (FACScan; Becton Dickinson; Franklin Lakes, NJ). A minimum of 10,000 cells was acquired and analyzed using software (Lysis II; Becton Dickinson). The lymphocyte gating to determinate the lymphocyte populations in BAL fluid and blood was made using forward- and side-scatter parameters and was controlled by using a combination of anti-CD45/CD14 monoclonal antibodies and CD3 + stained cells. The cells in BAL fluid and blood were stained according to identical protocols.

Lung Function

Spirometry was performed with a wedge spirometer (Vitalo-graph; Buckingham, UK) according to the American Thoracic Society recommendations. Bronchial histamine provocation was also performed as was described by Cockcroft. Airway resistance was measured in a constant-volume body plethysmo-graph (2000 TB; Cardio-Pulmonary Instruments; Houston, TX) Generic viagra nz. The subjects inhaled increased concentration of histamine chloride, starting at 0.063 mg/mL up to 64 mg/mL as maximal concentration, each step representing a doubling of the previous concentration. The provocation was stopped if airway resistance increased by 100%, or if stipulated highest histamine concentration (64 mg/mL) was reached, or if the challenge induced symptoms that made further histamine inhalations impossible. The outcome of histamine challenge was compared to the reference values of the clinic.

Statistical Analysis

Results are presented as median (25 to 75th percentiles) if not otherwise stated. Comparisons between the groups were assessed by the Kruskal-Wallis test followed by the Mann-Whitney U test when appropriate; p values < 0.05 were considered significant.