Characterization of a secretory phospholipase A2 in human bronchoalveolar lavage fluid.
Phospholipase A2 (PLA2) is a pivotal enzyme involved in the synthesis of the potent lipid inflammatory mediators platelet activating factor (PAF) and the eicosanoids. This study characterizes a PLA2 recovered in the bronchoalveolar lavage fluid (BALF) of healthy adult human subjects. Human BALF PLA2 exhibited characteristics of secretory PLA2s that include an activity that is acid stable, sensitive to reducing agents, and optimally requires millimolar calcium. BALF PLA2 showed marked selectivity for phosphatidylcholine containing arachidonic acid (AA) over linoleic or palmitic acids. Size exclusion chromatography showed the BALF PLA2 protein to be approximately 14 kDa in mass, consistent with it being a secretory form of PLA2. The biological significance of BALF PLA2 was tested by applying BALF concentrates to cultures of the human bronchial epithelial cell line BEAS 2B. Cultures of BEAS 2B cells treated with BALF concentrates released increased amounts of AA and produced higher levels of PAF. These data show that the lining fluid of the human respiratory tract normally contains a secretory PLA2, which may be involved in the formation of lipid inflammatory mediators in normal and pathophysiologic states in the lung.
Samet
J M
JM
Center for Environmental Medicine and Lung Biology, University of North Carolina, Chapel Hill 27599-7310, USA.
Madden
M C
MC
Fonteh
A N
AN
D000293:Adolescent; D000328:Adult; D016718:Arachidonic Acid / Q000032:analysis / Q000378:metabolism; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry*; D002118:Calcium / Q000032:analysis; D002460:Cell Line / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D008168:Lung / Q000201:enzymology* / Q000378:metabolism; D010741:Phospholipases A / Q000032:analysis / Q000378:metabolism*; D054467:Phospholipases A2; D010972:Platelet Activating Factor / Q000032:analysis / Q000378:metabolism
D010972:Platelet Activating Factor; D016718:Arachidonic Acid; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium