Published online before print May 3, 2004

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© by The Society for Leukocyte Biology
Journal of Leukocyte Biology, doi:10.1189/jlb.1003453

Received for publication October 1, 2003.
Revised March 10, 2004.
Accepted for publication March 13, 2004.

Article

Cytosolic phospholipase A₂ is responsible for prostaglandin E₂ and leukotriene B₄ formation in phagocyte-like PLB-985 cells: studies of differentiated cPLA₂-deficient PLB-985 cells

I. Furstenberg Liberty ^*, L. Raichel ^*, Z. Hazan-Eitan ^*, I. Pessach ^*, N. Hadad ^*, F. Schlaeffer , and R. Levy ^*@

Infectious Diseases Laboratory, Departments of *Clinical Biochemistry and Internal Medicine, Faculty of Health Sciences, Ben-Gurion University of the Negev and Soroka Medical Center, Beer Sheva, Israel

^@ To whom correspondence should be addressed. E-mail: ral{at}bgumail.bgu.ac.il.

Abstract

Our previously established model of cytosolic phospholipase A₂ (cPLA₂)-deficient, differentiated PLB-985 cells (PLB-D cells) was used to determine the physiological role of cPLA₂ in eicosanoid production. Parent PLB-985 (PLB) cells and PLB-D cells were differentiated toward the monocyte or granulocyte lineages using 5 x 10^-8 M 1,25 dihydroxyvitamin D₃ or 1.25% dimethyl sulfoxide, respectively. Parent monocyte- or granulocyte-like PLB cells released prostaglandin E₂ (PGE₂) when stimulated by ionomycin, A23187, opsonized zymosan, phorbol 12-myristate 13-acetate, or formyl-Met-Leu-Phe (fMLP), and monocyte- or granulocyte-like PLB-D cells did not release PGE₂ with any of the agonists. The kinetics of cPLA₂ translocation to nuclear fractions in monocyte-like PLB cells stimulated with fMLP or ionomycin was in correlation with the kinetics of PGE₂ production. Granulocyte-like PLB cells, but not granulocyte-like PLB-D cells, secreted leukotriene B₄ (LTB₄) after stimulation with ionomycin or A23187. Preincubation of monocyte-like parent PLB cells with 100 ng/ml lipopolysaccharide (LPS) for 16 h enhanced stimulated PGE₂ production, which is in correlation with the increased levels of cPLA₂ detected in these cells. LPS preincubation was less potent in increasing PGE₂ and LTB₄ secretion and did not affect cPLA₂ expression in granulocyte-like PLB cells, which may be a result of their lower levels of surface LPS receptor expression. LPS had no effect on monocyte- or granulocyte-like PLB-D cells. The lack of eicosanoid formation in stimulated, differentiated cPLA₂-deficient PLB cells indicates that cPLA₂ contributes to stimulated eicosanoid formation in monocyte- and granulocyte-like PLB cells.

Key Words: PGE₂ • LTB₄

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