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Originally published In Press as doi:10.1074/jbc.M006690200 on November 27, 2000

J. Biol. Chem., Vol. 276, Issue 8, 5467-5475, February 23, 2001
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Prostaglandin Levels in Stimulated Macrophages Are Controlled by Phospholipase A2-activating Protein and by Activation of Phospholipase C and D*

Deborah A. RibardoDagger , Sheila E. Crowe§, Kristine R. Kuhl, Johnny W. Peterson, and Ashok K. Chopra

From the Department of Microbiology and Immunology and § Internal Medicine, University of Texas Medical Branch, Galveston, Texas 77555-1070

Prostaglandins (PG), which are responsible for a large array of biological functions in eukaryotic cells, are produced from arachidonic acid by phospholipases and cyclooxygenase enzymes COX-1 and COX-2. We demonstrated that PG levels in cells were partly controlled by a regulatory protein, phospholipase A2 (PLA2)-activating protein (PLAA). Treatment of murine macrophages with lipopolysaccharide, interleukin-1beta , and tumor necrosis factor-alpha increased PLAA levels at early time points (2-30 min), which correlated with an up-regulation in cytosolic PLA2 and PGE2 levels. Both COX-2 and secretory PLA2 were also increased in lipopolysaccharide-stimulated macrophages, however, at later time points of 4-24 h. The role of PLAA in eicosanoid formation in macrophages was confirmed by the use of an antisense plaa oligonucleotide. Within amino acid residues 503-538, PLAA exhibited homology with melittin, and increased PGE2 production was noted in macrophages stimulated with melittin. In addition to PLA2, we demonstrated that activation of phospholipase C and D significantly controlled PGE2 production. Finally, increased antigen levels of PLAA, COX-2, and phospholipases were demonstrated in biopsy specimens from patients with varying amounts of intestinal mucosal inflammation, which corresponded to increased levels of phospholipase activity. These results could provide a basis for the development of new therapeutic tools to control inflammation.


* This work was supported in part by a grant from the Crohn's and Colitis Foundation of America and American Heart Association (to A. K. C.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Supported by The James W. McLaughlin Predoctoral Fellowship Fund.

To whom correspondence should be addressed: Dept. of Microbiology and Immunology, 301 University Blvd., University of Texas Medical Branch, Galveston, TX 77555-1070. Tel.: 409-747-0578; Fax: 409-747-6869; E-mail: achopra@utmb.edu.


Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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