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The Journal of Immunology, 2000, 165: 6504-6510.
Copyright © 2000 by The American Association of Immunologists

Delayed Onset of Inflammation in Protease-Activated Receptor-2-Deficient Mice1

Jonathan R. Lindner*, Mark L. Kahn2,{ddagger}, Shaun R. Coughlin{ddagger}, Gilberto R. Sambrano{ddagger}, Eric Schauble§, Daniel Bernstein§, Dan Foy{dagger}, Ali Hafezi-Moghadam{dagger} and Klaus Ley3,{dagger}

* Cardiovascular Division and {dagger} Department of Biomedical Engineering, University of Virginia School of Medicine, Charlottesville, VA 22908; {ddagger} Cardiovascular Research Institute, University of California, San Francisco, CA 94143; and § Pediatric Cardiology, Stanford University Medical Center, Stanford, CA 94305

Endothelial surface expression of P-selectin and subsequent leukocyte rolling in venules can be induced by mast cell-derived histamine and binding of thrombin to protease-activated receptor-1 (PAR1). We hypothesized that activation of endothelial PAR2 by mast cell tryptase or other proteases also contributes to inflammatory responses. Leukocyte rolling flux and rolling velocity were assessed by intravital microscopy of the cremaster muscles of wild-type mice following perivenular micropipette injections of a control (LSIGRL) or PAR2-activating (SLIGRL) oligopeptide. Injection of SLIGRL increased mean rolling leukocyte flux fraction from 34 ± 11 to 71 ± 24% (p < 0.05) and decreased mean rolling velocity from 63 ± 29 to 32 ± 2 µm/s (p < 0.05). No significant changes occurred with control peptide injection. To further evaluate the role of PAR2 in inflammatory responses, PAR2-deficient mice were generated by gene targeting and homologous recombination. Perivenular injections of SLIGRL resulted in only a small increase in rolling leukocyte flux fraction (from 21 ± 8 to 30 ± 2%) and no change in rolling velocity. Leukocyte rolling after surgical trauma was assessed in 9 PAR2-deficient and 12 wild-type mice. Early (0–15 min) after surgical trauma, the mean leukocyte rolling flux fraction was lower (10 ± 3 vs 30 ± 6%, p < 0.05) and mean rolling velocity was higher (67 ± 46 vs 52 ± 36 µm/s, p < 0.01) in PAR2-deficient compared with control mice. The defect in leukocyte rolling in PAR2-deficient mice did not persist past 30 min following surgical trauma. These results indicate that activation of PAR2 produces microvascular inflammation by rapid induction of P-selectin-mediated leukocyte rolling. In the absence of PAR2, the onset of inflammation is delayed.




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