Effects of Platelet Activating Factor on Various Physiological Parameters of Neutrophils, Alveolar Macrophages, and Alveolar Type II Cells (5e24da3c-a90e-4d19-95ee-e3e898aac14c)

"INTRODCJCTJONPlatelet activating factor (PAF) is a glycerophospholipid (1-0-alkyl-2-acctyl-sn-glycerol-3-phosphoryl choline) which has been shown to mediate a broad range of biological activities. 1·3 Its pulmonary actions include contraction of pulmonary tissue,4 secretion of leukotrienes from leukocytcs,5 airway constriction,6 pulmonary edema,7 and enhanced migration of neutrophils into the airspaces of thelungs.8-9PAF can be released from several different cell types, such as, basophils, neutrophils and alveolar macrophages, in response to a variety of particulates or membrane stimulants which include zymosan, calcium ionophore, phorbol esters, chemotactic agents, and endotoxin. 1~1 3 Therefore, PAF may play an important role in the development of pneumoconioses by mediating pulmonary responses of lung cells to a variety of occupational dusts. To investigate this possibility, we determined the effects of P AF on several physiological parameters of neutrophils, alveolar macrophages, and alveolar type Il epithelial cells.METHODSIsolation of CellsNeutrophils were isolated from human blood by dextran settling and centrifugal elutriation.14 Isolated neutrophils (93% pure) were resuspended in HEPES-buffered medium (145 mM NaCl, 5 mM KCI, 10 mM HEPES, 5 mM glucose, and 1 mM CaC12; pH = 7.4). Cell number and volume were determined with an electronic cell counter equipped with a sizing attachment.Rat alveolar macrophages were obtained by pulmonary lavage with ea2+, Mg2+-free Hanks balanced salts solution. 15 Alveolar macrophages (94% pure) were resuspended in HEPES-buffered medium, counted, and sized electronically.Rat alveolar type Il cells were isolated by enzymatic digestion for 35 minutes at 37°C with 40 µ/ml type I elastasc and 0.1 % collagenase and purified by centrifugal elutriation. 16.17 Type Il cells (92 % pure) were resuspended in HEPES-buffered medium for measurement of membrane potential, oxygen consumption, and trypan blue exclusion. Type Il pneumocytcs were resuspended in 0.1 M NaCl plus 0.05 M HEPES (pH = 7.8) to measure cytochrome P450-dependent activities and aggregation. Cell size and number were determined electronically."