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A more recent version of this article appeared on March 10, 2006
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Papers In Press, published online ahead of print January 1, 2006
J. Biol. Chem, 10.1074/jbc.M508314200
Submitted on July 29, 2005
Accepted on January 1, 2006

Group V secretory phospholipase A2 translocates to the phagosome after zymosan stimulation of mouse peritoneal macrophages and regulates phagocytosis

Barbara Balestrieri, Victor W. Hsu, Huiya Gilbert, Christina C. Leslie, Won K. Han, Joseph V. Bonventre, and Jonathan P. Arm

Division of Rheumatology Immunology and Allergy, Brigham and Women's Hospital, Boston, MA 02115

Corresponding Author: jarm{at}rics.bwh.harvard.edu

We have previously reported that group V secretory phospholipase A2 (sPLA2) amplifies the action of cytosolic phospholipase A2 (cPLA2) alpha in regulating eicosanoid biosynthesis by mouse peritoneal macrophages stimulated with zymosan (J Biol Chem; 2004: 279:16488-94). To further understand the role of group V sPLA2 we studied its localization in resting mouse peritoneal macrophages before and after stimulation with zymosan and the effect of deletion of the gene encoding group V sPLA2 on phagocytosis of zymosan. We report that group V sPLA2 is present in the Golgi apparatus and recycling endosome in the juxtanuclear region of resting peritoneal macrophages. Upon ingestion of zymosan by mouse peritoneal macrophages, group V sPLA2 is recruited to the phagosome. There it co-localizes with cPLA2alpha , 5-lipoxygenase, 5-lipoxygenase activating protein, and leukotriene C4 synthase. Using immunostaining for the cysteinyl leukotrienes in carbodiimide-fixed cells we show, for the first time, that the phagosome is a site of cysteinyl leukotriene formation. Furthermore, peritoneal macrophages from group V sPLA2-null mice demonstrated a >50% attenuation in phagocytosis of zymosan particles, which was restored by adenoviral expression of group V sPLA2 but not group IIA sPLA2. These data demonstrate that group V sPLA2 contributes to the innate immune response both through regulation of eicosanoid generation in response to a phagocytic stimulus and also as a component of the phagocytic machinery.


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