The calcium binding protein S100A9 is essential for pancreatic leukocyte infiltration and induces disruption of cell–cell contacts

Leukocyte infiltration is an early and critical event in the development of acute pancreatitis. However, the mechanism of leukocyte transmigration into the pancreas and the function of leukocytes in initiating acute pancreatitis are still poorly understood. Here, we studied the role of S100A9 (MRP14), a calcium binding protein specifically released by polymorph nuclear leukocytes (PMN), in the course of acute experimental pancreatitis. Acute pancreatitis was induced by repeated supramaximal caerulein injections in S100A9 deficient or S100A9 wild‐type mice. We then determined S100A9 expression, trypsinogen activation peptide (TAP) levels, serum amylase and lipase activities, and tissue myeloperoxidase (MPO) activity. Cell–cell contact dissociation was analyzed in vitro with biovolume measurements of isolated acini after incubation with purified S100A8/A9 heterodimers, and in vivo as measurement of Evans Blue extravasation after intravenous application of S100A8/A9. Pancreatitis induced increased levels of S100A9 in the pancreas. However, infiltration of leukocytes and MPO activity in the lungs and pancreas during acute pancreatitis was decreased in S100A9‐deficient mice and associated with significantly lower serum amylase and lipase activities as well as reduced intrapancreatic TAP‐levels. Incubation of isolated pancreatic acini with purified S100A8/A9‐heterodimers resulted in a rapid dissociation of acinar cell–cell contacts which was highly calcium‐dependent. Consistent with these findings, in vivo application of S100A8/A9 in mice was in itself sufficient to induce pancreatic cell–cell contract dissociation as indicated by Evans Blue extravasation. These data show that the degree of intrapancreatic trypsinogen activation is influenced by the extent of leukocyte infiltration into the pancreas which, in turn, depends on the presence of S100A9 that is secreted from PMN. S100A9 directly affects leukocyte tissue invasion and mediates cell contact dissociation via its calcium binding properties. J. Cell. Physiol. 216: 558–567, 2008. © 2008 Wiley‐Liss, Inc.

[1]  P. Tessier,et al.  S100A9 mediates neutrophil adhesion to fibronectin through activation of beta2 integrins. , 2007, Biochemical and biophysical research communications.

[2]  J. Palefsky,et al.  Oxidation of methionine 63 and 83 regulates the effect of S100A9 on the migration of neutrophils in vitro , 2007, Journal of leukocyte biology.

[3]  J. Palefsky,et al.  S100A8 Triggers Oxidation-sensitive Repulsion of Neutrophils , 2006, Journal of dental research.

[4]  A. Remppis,et al.  Increased proinflammatory endothelial response to S100A8/A9 after preactivation through advanced glycation end products , 2006 .

[5]  S. Pedigo,et al.  Calcium-dependent stability studies of domains 1 and 2 of epithelial cadherin. , 2005, Biochemistry.

[6]  J. Mayerle,et al.  Extracellular cleavage of E-cadherin by leukocyte elastase during acute experimental pancreatitis in rats. , 2005, Gastroenterology.

[7]  Jürgen Schnekenburger,et al.  Protein tyrosine phosphatase κ and SHP-1 are involved in the regulation of cell-cell contacts at adherens junctions in the exocrine pancreas , 2005, Gut.

[8]  R. Schmid.  Pathophysiology of Acute Pancreatitis , 2005, Digestion.

[9]  M. Bhatia,et al.  Pathophysiology of Acute Pancreatitis , 2005, Pancreatology.

[10]  D. Foell,et al.  Myeloid-related proteins 8 and 14 induce a specific inflammatory response in human microvascular endothelial cells. , 2005, Blood.

[11]  I. Thorey,et al.  MRP8 and MRP14 control microtubule reorganization during transendothelial migration of phagocytes. , 2004, Blood.

[12]  W. Baumgartner,et al.  Ca2+ Dependency of N-Cadherin Function Probed by Laser Tweezer and Atomic Force Microscopy , 2003, The Journal of Neuroscience.

[13]  J. Mayerle,et al.  Pathophysiology of Alcohol-Induced Pancreatitis , 2003, Pancreas.

[14]  P. Rouleau,et al.  Blockade of S100A8 and S100A9 Suppresses Neutrophil Migration in Response to Lipopolysaccharide 1 , 2003, The Journal of Immunology.

[15]  W. Nacken,et al.  S100A9/S100A8: Myeloid representatives of the S100 protein family as prominent players in innate immunity , 2003, Microscopy research and technique.

[16]  N. Hogg,et al.  Myeloid Cell Function in MRP-14 (S100A9) Null Mice , 2003, Molecular and Cellular Biology.

[17]  P. Rouleau,et al.  Proinflammatory Activities of S100: Proteins S100A8, S100A9, and S100A8/A9 Induce Neutrophil Chemotaxis and Adhesion 1 , 2003, The Journal of Immunology.

[18]  W. Nacken,et al.  Loss of S100A9 (MRP14) Results in Reduced Interleukin-8-Induced CD11b Surface Expression, a Polarized Microfilament System, and Diminished Responsiveness to Chemoattractants In Vitro , 2003, Molecular and Cellular Biology.

[19]  D. Vestweber.  Regulation of endothelial cell contacts during leukocyte extravasation. , 2002, Current opinion in cell biology.

[20]  S. Holland,et al.  Neutrophils and NADPH oxidase mediate intrapancreatic trypsin activation in murine experimental acute pancreatitis. , 2002, Gastroenterology.

[21]  M. Lerch,et al.  Trypsin activity is not involved in premature, intrapancreatic trypsinogen activation. , 2002, American journal of physiology. Gastrointestinal and liver physiology.

[22]  M. Aurrand-Lions,et al.  The last molecular fortress in leukocyte trans-endothelial migration , 2002, Nature Immunology.

[23]  M. Büchler,et al.  Role of activation peptides from pancreatic proenzymes in the diagnosis and prognosis of acute pancreatitis. , 2002, Surgery.

[24]  A. Varki,et al.  Two Proteins Modulating Transendothelial Migration of Leukocytes Recognize Novel Carboxylated Glycans on Endothelial Cells1 , 2001, The Journal of Immunology.

[25]  H Lippert,et al.  Role of cathepsin B in intracellular trypsinogen activation and the onset of acute pancreatitis. , 2000, The Journal of clinical investigation.

[26]  M. Lerch,et al.  The role of intracellular calcium signaling in premature protease activation and the onset of pancreatitis. , 2000, The American journal of pathology.

[27]  H Schindler,et al.  Cadherin interaction probed by atomic force microscopy. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[28]  C. Kerkhoff,et al.  The Regulatory Role of MRP8 (S100A8) and MRP14 (S100A9) in the Transendothelial Migration of Human Leukocytes , 2000, Pathobiology.

[29]  C. Kerkhoff,et al.  The Two Calcium-binding Proteins, S100A8 and S100A9, Are Involved in the Metabolism of Arachidonic acid in Human Neutrophils* , 1999, The Journal of Biological Chemistry.

[30]  D. Hume,et al.  A null mutation in the inflammation-associated S100 protein S100A8 causes early resorption of the mouse embryo. , 1999, Journal of immunology.

[31]  A. Saluja,et al.  The role of intercellular adhesion molecule 1 and neutrophils in acute pancreatitis and pancreatitis-associated lung injury. , 1999, Gastroenterology.

[32]  C. Kerkhoff,et al.  Novel insights into structure and function of MRP8 (S100A8) and MRP14 (S100A9). , 1998, Biochimica et biophysica acta.

[33]  N. Hogg,et al.  The human S100 protein MRP-14 is a novel activator of the beta 2 integrin Mac-1 on neutrophils. , 1998, Journal of immunology.

[34]  S. Pandol,et al.  Pancreatic acinar cells produce, release, and respond to tumor necrosis factor-alpha. Role in regulating cell death and pancreatitis. , 1997, The Journal of clinical investigation.

[35]  M. Hartmann,et al.  Myeloid-related Protein (MRP) 8 and MRP14, Calcium-binding Proteins of the S100 Family, Are Secreted by Activated Monocytes via a Novel, Tubulin-dependent Pathway* , 1997, The Journal of Biological Chemistry.

[36]  P. Braquet,et al.  The role of neutrophils and platelet-activating factor in mediating experimental pancreatitis. , 1996, Gastroenterology.

[37]  J. Engel,et al.  Conformational changes of the recombinant extracellular domain of E-cadherin upon calcium binding. , 1994, European journal of biochemistry.

[38]  M. Lerch,et al.  The effect of chloroquine administration on two experimental models of acute pancreatitis. , 1993, Gastroenterology.

[39]  B. Austen,et al.  Development of radioimmunoassays for free tetra-L-aspartyl-L-lysine trypsinogen activation peptides (TAP). , 1988, Journal of immunological methods.

[40]  C. Sorg,et al.  Two calcium-binding proteins associated with specific stages of myeloid cell differentiation are expressed by subsets of macrophages in inflammatory tissues. , 1988, Clinical and experimental immunology.

[41]  C. Niederau,et al.  Diagnosis of chronic pancreatitis. , 1985, Gastroenterology.

[42]  L. Ferrell,et al.  Caerulein-induced acute necrotizing pancreatitis in mice: protective effects of proglumide, benzotript, and secretin. , 1985, Gastroenterology.

[43]  A. Saria,et al.  Evans blue fluorescence: quantitative and morphological evaluation of vascular permeability in animal tissues , 1983, Journal of Neuroscience Methods.

[44]  M. Korc,et al.  Action of secretagogues on a new preparation of functionally intact, isolated pancreatic acini. , 1978, The American journal of physiology.

[45]  P. Rouleau,et al.  Proinflammatory Activities of S 100 : Proteins S 100 A 8 , S 100 A 9 , and S 100 A 8 / A 9 Induce Neutrophil Chemotaxis and Adhesion 1 , 2003 .

[46]  T. Keck,et al.  Matrix metalloproteinase-9 promotes neutrophil migration and alveolar capillary leakage in pancreatitis-associated lung injury in the rat. , 2002, Gastroenterology.