Characterization of phospholipase A2 from human nasal lavage.
A secreted form of phospholipase A2 (PLA2) has been implicated in inflammatory disorders such as rheumatoid arthritis and sepsis. To determine if PLA2 may also play a role in allergic rhinitis, we have measured enzymic activity in nasal lavage from allergic subjects. Enhanced activity of PLA2 in the lavage was observed following nasal challenge with antigen or histamine. The PLA2 in the nasal lavage was partially purified by acid extraction, size exclusion chromatography, and ion exchange chromatography. The partially purified enzyme from nasal lavage was subsequently compared to a recombinant form of human PLA2 identified in synovial fluid from arthritic patients. The two enzymes showed similar molecular weights (15 to 16 kD) on SDS-PAGE, and both reacted with a rabbit polyclonal antiserum raised to a galactokinase-PLA2 fusion protein. The enzymatic activities of the two PLA2s were indistinguishable when compared for ionic dependence, substrate selectivity, and sensitivity to inhibitors. These results suggest that the PLA2 induced in nasal lavage in response to challenge by antigen is very similar to the extracellular PLA2 found in synovial fluid from subjects with rheumatoid arthritis and may play a role in the inflammatory processes associated with allergic rhinitis.
Department of Pharmacology, SmithKline Beecham Pharmaceuticals, King of Prussia, Pennsylvania 19406.
J M
JM
Stadel
K
K
Hoyle
R M
RM
Naclerio
A
A
Roshak
F H
FH
Chilton
D000293:Adolescent; D000328:Adult; D000941:Antigens; D001168:Arthritis / Q000201:enzymology; D004229:Dithiothreitol / Q000494:pharmacology; D005260:Female; D006801:Humans; D008297:Male; D017817:Nasal Lavage Fluid / Q000737:chemistry* / Q000276:immunology; D009299:Nasal Provocation Tests; D010741:Phospholipases A / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D054467:Phospholipases A2; D011993:Recombinant Fusion Proteins / Q000276:immunology; D011994:Recombinant Proteins; D006255:Rhinitis, Allergic, Seasonal / Q000201:enzymology*; D013582:Synovial Fluid / Q000201:enzymology
D000941:Antigens; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D010741:Phospholipases A; D054467:Phospholipases A2; D004229:Dithiothreitol