All patients received 200 mg of erythromycin stearate orally 3 times per day for more than 6 months until BAL was repeated. The patients were instructed at each outpatient attendance to take one tablet every 8 hours, at home, and they were given a 4-week supply. They were instructed to bring back any remaining tablets at each visit to that the numbers of remaining tablets could be checked. None of the patients received corticosteroids or antibiotics other than EM for the duration of this study read only.
The patients were premedicated intramuscularly with atropine (0.5 mg) and local anesthesia was produced with 2 percent lidocaine; airway examination was then carried out with a flexible fiberoptic bronchoscope (Olympas BF-P20 type; Olympus Corp, New Hyde Park, NY).
The bronchoscope was wedged securely into the subsegmental bronchus of the right middle lobe and 150 ml of sterile 0.9 percent NaCl at 37 °C was infused in three 50-ml aliquots, and gently aspirated immediately after each infusion. The recovered lavage fluid was pooled, passed through a double layer of gauze to remove gross mucus, then centrifuged. A differential cell count was performed on the pellet, and the supernatant was frozen at —80 °C until used.
Neutrophils for the chemotactic assay were isolated from a normal healthy volunteer by mono-poly resolving medium (M-PRM; Flow Laboratories, Irvine, Scotland) density gradient centrifugation. The neutrophils were suspended, at a density of 3X106 cells per ml, in Hanks’ solution (HBSS, Gibco), pH 7.2, containing 0.1 percent bovine serum albumin (BSA).
More than 98 percent of the cells were neutrophils; cell viability was in excess of 95 percent, as determined by trypan blue exclusion. Neutrophil chemotactic activity was assessed using a 48-well microchemotaxis chamber (Neuroprobe Inc, Bethesda, Md) as described elsewhere. To each of the bottom wells, 25-ц\ aliquots of samples were added. A polycarbonate filter sheet (3-/Ш1 pores), not containing polyvinylpyrrolidone (NFB3-PVPF, Costar Corp, Cambridge, Mass), was placed on top of the wells in the bottom plate.