Published online before print April 22, 2009
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* Department of Pediatrics, University of Colorado Denver, Aurora, Colorado, USA; and
Department of Pediatrics, National Jewish Medical and Research Center, Denver, Colorado, USA
2. Correspondence: University of Colorado Denver, Pediatrics, Mail Stop 8302, RC-1 North, 12800 E. 19th Ave., Room P18-4400, Aurora, CO 80045, USA. E-mail: doug.graham{at}ucdenver.edu
ABSTRACT
The MerTK plays several important roles in normal macrophage physiology, including regulation of cytokine secretion and clearance of apoptotic cells. Mer signaling in other cell types, including malignant cells that ectopically overexpress the RTK, leads to downstream prosurvival pathway activation. We explored the hypothesis that Mer has a prosurvival role in macrophages exposed to oxidative stress. H2O2 treatment of peritoneal exudate murine macrophages and J774 cells rapidly stimulated Mer phosphorylation in a concentration-dependent manner. Mer phosphorylation was dependent on the ligand Gas6, as treatment with warfarin or MerFc (a fusion protein of the extracellular domain of Mer and the Fc portion of human Ig), inhibitors of Gas6 activity, blocked H2O2-mediated activation of Mer. Antiapoptotic signals including pAkt and pErk 1/2 were increased dramatically (threefold and 4.5-fold, respectively) in WT Mer-positive macrophages compared with Mer KO macrophages stimulated with H2O2. In a consistent manner, Mer expression led to decreased cleavage of proapoptotic indicators PARP and Caspase-3. Furthermore, Mer provided up to twofold enhanced cellular survival to primary macrophages exposed to H2O2. These data represent the first report of Mer activation in response to oxidative stress and demonstrate the ability of Mer RTK to promote macrophage survival in disease states that involve an oxidative stress environment.
Key Words: hydrogen peroxide receptor leukocyte antiapoptotic signaling