The Cleavage of Neutrophil Leukosialin (CD43) by Cathepsin G Releases Its Extracellular Domain and Triggers Its Intramembrane Proteolysis by Presenilin/γ-Secretase

Abstract

The highly negatively charged membrane sialoglycoprotein leukosialin, CD43, is shed during neutrophil activation. This is generally thought to enhance cell adhesion. We here describe two novel consequences of this shedding, during neutrophil activation by phorbol esters or by chemoattractants after TNF-α priming. CD43 proteolysis was investigated by Western blotting, using a polyclonal antibody to CD43 intracellular domain. Our data emphasize the importance of a juxtamembranous cleavage of about 50% of membrane CD43 molecules by cathepsin G. Indeed, it is inhibited by α1-antichymotrypsin and cathepsin G inhibitor I and is reproduced by exogenous purified cathepsin G. The resulting membrane-anchored C-terminal fragment, CD43-CTF, becomes susceptible to presenilin/γ-secretase, which releases CD43 intracytoplasmic domain: preincubation with three different γ-secretase inhibitors, before PMN treatment by agonists or by purified cathepsin G, results in the accumulation of CD43-CTF. Because CD43 binds E-selectin, we also investigated the effect of the soluble extracellular domain CD43s, released by cathepsin G juxtamembranous cleavage, on neutrophil adhesion to endothelial cells. A recombinant CD43s-Fc fusion protein inhibited neutrophil E selectindependent adhesion to endothelial cells under flow conditions, while it had no effect on neutrophil static adhesion. We thus propose that, in addition to its potential pro-adhesive role, CD43 proteolysis results in: (i) the release, by cathepsin G, of CD43 extracellular domain, able to inhibit the adhesion of flowing neutrophils on endothelial cells and thus to participate to the natural control of inflammation; (ii) the release and/or the clearance, by presenilin/γ-secretase, of CD43 intracellular domain, thereby regulating CD43-mediated signaling. The highly negatively charged membrane sialoglycoprotein leukosialin, CD43, is shed during neutrophil activation. This is generally thought to enhance cell adhesion. We here describe two novel consequences of this shedding, during neutrophil activation by phorbol esters or by chemoattractants after TNF-α priming. CD43 proteolysis was investigated by Western blotting, using a polyclonal antibody to CD43 intracellular domain. Our data emphasize the importance of a juxtamembranous cleavage of about 50% of membrane CD43 molecules by cathepsin G. Indeed, it is inhibited by α1-antichymotrypsin and cathepsin G inhibitor I and is reproduced by exogenous purified cathepsin G. The resulting membrane-anchored C-terminal fragment, CD43-CTF, becomes susceptible to presenilin/γ-secretase, which releases CD43 intracytoplasmic domain: preincubation with three different γ-secretase inhibitors, before PMN treatment by agonists or by purified cathepsin G, results in the accumulation of CD43-CTF. Because CD43 binds E-selectin, we also investigated the effect of the soluble extracellular domain CD43s, released by cathepsin G juxtamembranous cleavage, on neutrophil adhesion to endothelial cells. A recombinant CD43s-Fc fusion protein inhibited neutrophil E selectindependent adhesion to endothelial cells under flow conditions, while it had no effect on neutrophil static adhesion. We thus propose that, in addition to its potential pro-adhesive role, CD43 proteolysis results in: (i) the release, by cathepsin G, of CD43 extracellular domain, able to inhibit the adhesion of flowing neutrophils on endothelial cells and thus to participate to the natural control of inflammation; (ii) the release and/or the clearance, by presenilin/γ-secretase, of CD43 intracellular domain, thereby regulating CD43-mediated signaling. The regulated proteolysis of transmembrane proteins represents an important mechanism of cell functions modulation. The inflammation resolution involves, for example, the shedding of cytokine receptors and adhesion molecules, which down-regulates leukocyte adhesion to endothelium. These regulations result from a decreased membrane expression of receptors and/or from the release of soluble fragments, competing with their membrane counterparts. The majority of shed proteins identified to date are cleaved by metalloproteinases or by neutrophil-derived serine proteases (1Garton K.J. Gough P.J. Raines E.W. J. Leuk. Biol. 2006; 79: 1105-1116Crossref PubMed Scopus (191) Google Scholar). Leukosialin, CD43, is the predominant cell surface sialoprotein of leukocytes (2Nathan C. Xie Q.W. Halbwachs-Mecarelli L. Jin W.W. J. Cell Biol. 1993; 122: 243-256Crossref PubMed Scopus (115) Google Scholar) and has both anti-adhesive and adhesive properties. Its function has been mainly studied on lymphocytes, where CD43 behaves both as a negative regulator of T cell proliferation and adhesion and as a positive regulator of memory T cell trafficking (3Manjunath N. Correa M. Ardman M. Ardman B. Nature. 1995; 377: 535-538Crossref PubMed Scopus (194) Google Scholar, 4Mody P.D. Cannon J.L. Bandukwala H.S. Blaine K.M. Schilling A.B. Swier K. Sperling A.I. Blood. 2007; 110: 2974-2982Crossref PubMed Scopus (21) Google Scholar). Although its expression is normally restricted to leukocytes, CD43 is present on colon carcinomas and on several nonhematopoietic cell lines (5Sikut R. Nilsson O. Baeckstrom D. Hansson G.C. Biochem. Biophys. Res. Commun. 1997; 238: 612-616Crossref PubMed Scopus (28) Google Scholar, 6Fernandez-Rodriguez J. Andersson C.X. Laos S. Baeckstrom D. Sikut A. Sikut R. Hansson G.C. Tumour Biol. 2002; 23: 193-201Crossref PubMed Scopus (27) Google Scholar). In these cell lines, CD43 is processed by a presenilin/γ-secretase-mediated regulated intramembrane proteolysis (RIP) 3The abbreviations used are:RIPregulated intramembrane proteolysisα1PIα1-proteinase inhibitor (α1-antitrypsin)a1-CTα1 anti-chymotrypsinBSAbovine serum albuminCD43-CTFCD43 C-terminal fragmentCD43ssoluble CD43fMLPN-formyl-l-methionyl-l-leucyl-l-phenylalanineGalgpgalactoglycoproteinHBSSHanks balanced solutionHUVEChuman umbilical vein endothelial cellsMMPmatrix metalloproteinaseNEneutrophil elastasePMAphorbol myristate acetatePMNpolymorphonuclear neutrophilPR3proteinase 3TNFtumor necrosis factorFITCfluorescein isothiocyanatePSGLP-selectin glycoprotein ligandmAbmonoclonal antibodyPSpresenilin. (7Andersson C.X. Fernandez-Rodriguez J. Laos S. Baeckstrom D. Haass C. Hansson G.C. Biochem. J. 2005; 387: 377-384Crossref PubMed Scopus (39) Google Scholar). RIP refers to a sequential proteolysis of various type I membrane proteins, including the amyloid precursor protein of Alzheimer disease, the Notch receptor, CD44, and E-cadherin (8Selkoe D.J. Wolfe M.S. Cell. 2007; 131: 215-221Abstract Full Text Full Text PDF PubMed Scopus (315) Google Scholar). RIP is initiated usually by a metalloproteinase, which induces the shedding of the receptor ectodomain, followed by an intramembrane processing of the cell-bound fragment by a PS/γ secretase. This releases the receptor intracellular domain in the cytoplasm. In the case of CD43 in cancer cell lines, this domain translocates in the nucleus and causes the up-regulation of various genes (9Andersson C.X. Fernandez-Rodriguez J. Laos S. Sikut R. Sikut A. Baeckstrom D. Hansson G.C. Biochem. Biophys. Res. Commun. 2004; 316: 12-17Crossref PubMed Scopus (24) Google Scholar). regulated intramembrane proteolysis α1-proteinase inhibitor (α1-antitrypsin) α1 anti-chymotrypsin bovine serum albumin CD43 C-terminal fragment soluble CD43 N-formyl-l-methionyl-l-leucyl-l-phenylalanine galactoglycoprotein Hanks balanced solution human umbilical vein endothelial cells matrix metalloproteinase neutrophil elastase phorbol myristate acetate polymorphonuclear neutrophil proteinase 3 tumor necrosis factor fluorescein isothiocyanate P-selectin glycoprotein ligand monoclonal antibody presenilin. CD43 has been recently described as a ligand for E-selectin (10Fuhlbrigge R.C. King S.L. Sackstein R. Kupper T.S. Blood. 2006; 107: 1421-1426Crossref PubMed Scopus (95) Google Scholar, 11Matsumoto M. Atarashi K. Umemoto E. Furukawa Y. Shigeta A. Miyasaka M. Hirata T. J. Immunol. 2005; 175: 8042-8050Crossref PubMed Scopus (101) Google Scholar). Its expression on polymorphonuclear neutrophils (PMN) is 10-fold higher than P-selectin glycoprotein ligand-1, PSGL-1, 4L. Halbwachs-Mecarelli, unpublished observation. the main leukocyte ligand for P-selectin, which also binds E-selectin. Despite CD43 abundance, data on its precise role in PMN responses are scarce. CD43 is shed during PMN activation (12Campanero M.R. Pulido R. Alonso J.L. Pivel J.P. Pimentel-Muinos F.X. Fresno M. Sanchez-Madrid F. Eur. J. Immunol. 1991; 21: 3045-3048Crossref PubMed Scopus (106) Google Scholar, 13Kuijpers T.W. Hoogerwerf M. Kuijpers K.C. Schwartz B.R. Harlan J.M. J. Immunol. 1992; 149: 998-1003PubMed Google Scholar, 14Rieu P. Porteu F. Bessou G. Lesavre P. Halbwachs-Mecarelli L. Eur. J. Immunol. 1992; 22: 3021-3026Crossref PubMed Scopus (67) Google Scholar, 15Remold-O'Donnell E. Parent D. J. Immunol. 1994; 152: 3595-3605PubMed Google Scholar) and during adhesion and spreading (2Nathan C. Xie Q.W. Halbwachs-Mecarelli L. Jin W.W. J. Cell Biol. 1993; 122: 243-256Crossref PubMed Scopus (115) Google Scholar, 16Lopez S. Seveau S. Lesavre P. Robinson M.K. Halbwachs-Mecarelli L. Cell Adhes. Commun. 1998; 5: 151-160Crossref PubMed Scopus (24) Google Scholar), together with E-selectin and J.L. J. Immunol. 1994; 152: Google Scholar, M. T. Blood. PubMed Scopus (67) Google Scholar). A soluble of CD43, CD43s, the extracellular domain, has been described in and identified to galactoglycoprotein K. R. T. J. K. 1992; PubMed Scopus Google Scholar). CD43 proteolysis on PMN was to serine proteases and The precise of the proteolysis mechanism was by the CD43 soluble on E. Parent D. J. Immunol. 1994; 152: 3595-3605PubMed Google Scholar) and an of neutrophil We a polyclonal antibody a recombinant CD43 intracellular domain to the proteolysis in the of data (i) the γ-secretase processing of CD43 in cancer (ii) the of CD43 as a leukocyte ligand for endothelial E-selectin. We here describe for the in human PMN by a cathepsin G and γ-secretase processing of CD43 with important and and from from and from The was as described P. C. M. Seveau S. J.M. Lesavre P. Halbwachs-Mecarelli L. J. Biol. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar). neutrophil elastase cathepsin G, and proteinase 3 purified as described P. Halbwachs-Mecarelli L. P. Lesavre P. M. J. Immunol. 1994; 152: Google Scholar). cathepsin G was also from and purified on protein from an human The CD43s-Fc fusion protein was in as described R. S. M. J. Biol. 1994; Full Text PDF PubMed Google Scholar) and purified on protein necrosis factor was from human was from cathepsin G inhibitor and from E from and from while and a from and from neutrophils and endothelial cells as described P. C. M. Seveau S. J.M. Lesavre P. Halbwachs-Mecarelli L. J. Biol. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar, J. PubMed Scopus Google Scholar) from and umbilical from in Hanks balanced solution with and bovine serum albumin In this has no effect on CD43 proteolysis (2Nathan C. Xie Q.W. Halbwachs-Mecarelli L. Jin W.W. J. Cell Biol. 1993; 122: 243-256Crossref PubMed Scopus (115) Google Scholar). on in with endothelial cell and and cells used cells with CD43 or with both CD43 and as described K. M. J. Biol. 1994; Full Text PDF PubMed Google Scholar), to enhance protein and Western neutrophils in and during their activation with phorbol myristate acetate or with TNF-α and N-formyl-l-methionyl-l-leucyl-l-phenylalanine as described in the The was by and to CD43 PMN for and to Western or flow as described P. C. M. Seveau S. J.M. Lesavre P. Halbwachs-Mecarelli L. J. Biol. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar). for PMN to in for with TNF-α and with PMN of with and to for cells by and PMN and with for with to PMN from endothelial cells. PMN by flow with a adhesion was as described P. A. J. O. N. D. J. 2006; PubMed Scopus Google Scholar), by with a in a flow a with which had been for with TNF-α and to the flow a of to two resulting in sequential of and PMN for the endothelial cells This was followed by a of with and and as described D.J. P. PubMed Scopus Google Scholar). The the of cells in three using a was as and CD43 by Western a E. Parent D. J. Immunol. 1994; 152: 3595-3605PubMed Google Scholar), we extracellular soluble CD43 to membrane charged or We thus an antibody to by Western the CD43 intracellular and membrane-anchored after The antibody was by Western with cell lines with CD43 and with resulting in to the and of CD43 K. M. J. Biol. 1994; Full Text PDF PubMed Google Scholar) of PMN it in to the soluble CD43 by of neutrophils E. Parent D. J. Immunol. 1994; 152: 3595-3605PubMed Google Scholar, L. Bessou G. Lesavre P. P. M. Blood. PubMed Google (i) a cell-bound resulting from A and to the soluble (ii) a cell-bound CD43 C-terminal fragment, CD43-CTF, of resulting from and to the soluble The of the was and to to the activation of PMN during the we had with the soluble L. Bessou G. Lesavre P. P. M. Blood. PubMed Google Scholar), it was on cell and was by the of and by the of in is the represents two or a resulting in by as the than the and was the main fragment after of PMN activation by TNF-α or CD43, as P. Porteu F. Bessou G. Lesavre P. Halbwachs-Mecarelli L. Eur. J. Immunol. 1992; 22: 3021-3026Crossref PubMed Scopus (67) Google Scholar), while with TNF-α for followed by of activation with in CD43 cleavage to the was present or CD43 in CD43 proteolysis the of the CD43 and the of CD43 by to a by TNF-α with This by was Although neutrophil activation in in or no of the as with control cells in with cells This the extracellular domain of CD43 from while the juxtamembranous cleavage is to of to the fragment, in for the CD43 of PMN a of CD43 activation in with and of in CD43 of CD43 proteolysis Because the main are α1-proteinase inhibitor and we the of these on CD43 Western to cleavage, the of a fragment and the of the is to inhibit the serine proteases released by PMN cathepsin G, and proteinase with the for cathepsin G elastase J. Biochem. PubMed Scopus Google Scholar). These results thus cathepsin G as a for the cleavage of the fragment a cell-bound of CD43 membrane after neutrophil with exogenous purified serine no cleavage by while elastase and cathepsin G in to and CD43 in and in with data E. Parent D. Blood. 1995; PubMed Google Scholar), Western of PMN with purified elastase in a to with also a cathepsin G a juxtamembranous cleavage, a fragment to the after PMN activation by in with cathepsin G results with two cathepsin G the cathepsin G inhibitor inhibited the of the resulting from activation We thus propose CD43 proteolysis (i) A and by and resulting in and fragments, (ii) a juxtamembranous by cathepsin G, the CD43-CTF. Our results also to followed by and thus is by of cancer an intramembrane cleavage of CD43 by has been described (7Andersson C.X. Fernandez-Rodriguez J. Laos S. Baeckstrom D. Haass C. Hansson G.C. Biochem. J. 2005; 387: 377-384Crossref PubMed Scopus (39) Google Scholar). We thus investigated a role of γ-secretase in CD43 processing in In intracytoplasmic are to to the nucleus or are by the is thus by the accumulation of membrane in the of γ-secretase The of three different of or during neutrophil activation by the of the on Western in the of γ-secretase inhibitors, the on Western from neutrophils by with γ-secretase in the of fragment thus the of a juxtamembranous cleavage, PMN activation by which was of by We intracytoplasmic resulting from this cell in to proteins before Western of in CD43 proteolysis is usually by matrix We thus the of various on CD43 CD43 cleavage was inhibited by metalloproteinase inhibitors, as and and various including and and the and inhibitor A and In the shedding of on the cells CD43 was inhibited by and also by the data J.L. J. Immunol. 1994; 152: Google Scholar) and an effect to metalloproteinase thus to in CD43 by as a of G thus cathepsin G the function of in the RIP processing and CD43 for γ-secretase in exogenous purified cathepsin G in a fragment, which was Indeed, the of the fragment, resulting from of cathepsin G, was by PMN preincubation with γ-secretase inhibitor E the of cathepsin G we these with two cathepsin G and the results the of CD43 released by exogenous elastase by E PMN treatment by cathepsin G, in the of the γ-secretase in the of the cathepsin G inhibitor by the inhibitor This result the cleavage of CD43 by purified cathepsin G, in the of PMN is to CD43 processing by of in the of had no effect on neutrophil by or by as by flow data had no effect on the with the data inhibited neutrophil adhesion to by or by of PMN to by CD43s, the of CD43 after Cell the effect of a recombinant on neutrophil adhesion to endothelial cells. the function of the natural was in which the for a to neutrophils K. M. J. Biol. 1994; Full Text PDF PubMed Google Scholar). We used on K. R. T. J. K. 1992; PubMed Scopus Google Scholar). a using and and with the recombinant in a of of in data Our results has no effect on PMN static adhesion to extracellular matrix data or to endothelial cells We with this adhesion is PMN adhesion to under flow conditions, by was also inhibited by with human purified from an as a control for the The was the with and In conditions, recombinant had no effect on PMN adhesion These results with the adhesion of PMN to endothelial cells under Our of CD43 resulting from neutrophil several E. Parent D. J. Immunol. 1994; 152: 3595-3605PubMed Google Scholar). Indeed, the of membrane-anchored C-terminal fragments, with are of described for soluble We CD43 proteolysis three or sequential in and A and from the membrane and a juxtamembranous B. The described as a cleavage E. Parent D. J. Immunol. 1994; 152: 3595-3605PubMed Google Scholar) to a protein domain to This cleavage was PMN in of and it with the cell This it is by proteases present on normally by or or on PMN membrane by the This cleavage was by or by and we it results in CD43 to with exogenous elastase L. Bessou G. Lesavre P. P. M. Blood. PubMed Google Scholar, E. Parent D. Blood. 1995; PubMed Google Scholar). In purified elastase results in a cleavage to the membrane and a A fragment was during PMN was in the of α1-antichymotrypsin which its These results that, during PMN CD43 A and from the cell membrane and resulting in cell-bound and The A is and The fragment to processed by the in The of as the extracellular CD43 domain released in the K. R. T. J. K. 1992; PubMed Scopus Google Scholar) it the juxtamembranous is in the shedding of This had been by the of a and to by E. Parent D. J. Immunol. 1994; 152: 3595-3605PubMed Google Scholar). Our to for the the of juxtamembranous which results in a membrane C-terminal fragment We also this cleavage is by also by as TNF-α followed by activation. In the the resulting is in the of γ-secretase inhibitors, as also during PMN we described a membrane-anchored CD43 fragment on P. C. M. Seveau S. J.M. Lesavre P. Halbwachs-Mecarelli L. J. Biol. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar). The juxtamembranous had been to result from an to serine or metalloproteinase E. Parent D. J. Immunol. 1994; 152: 3595-3605PubMed Google Scholar). We here that, which of a activation by or by together with while it is with TNF-α or This is of the of which PMN together with serine which are are cathepsin G and are to with a membrane protein as Our flow data elastase and cathepsin G result in CD43 shedding, while has no effect E. Parent D. Blood. 1995; PubMed Google Scholar). Western of CD43 proteolysis by cathepsin G, which had been investigated cathepsin G a for the juxtamembranous B. (i) both which cathepsin G elastase and a cathepsin G inhibitor inhibited CD43 proteolysis and the of the (ii) PMN treatment with purified cathepsin G from three different in a juxtamembranous cleavage, a to after PMN activation by the of CD43 E. S. A. PubMed Scopus Google Scholar) a cathepsin G K. M. J. Biol. Full Text PDF PubMed Google Scholar), from the transmembrane a cleavage release an extracellular soluble fragment with the C-terminal as galactoglycoprotein the soluble CD43 described in K. R. T. J. K. 1992; PubMed Scopus Google Scholar). The cleavage of CD43, resulting from neutrophil is thus to by cathepsin G. The of of CD43 proteolysis by or of elastase and cathepsin G, and different been (2Nathan C. Xie Q.W. Halbwachs-Mecarelli L. Jin W.W. J. Cell Biol. 1993; 122: 243-256Crossref PubMed Scopus (115) Google Scholar, 15Remold-O'Donnell E. Parent D. J. Immunol. 1994; 152: 3595-3605PubMed Google Scholar). The of proteases released by neutrophils and is to by PMN and C. J. Immunol. 2004; PubMed Scopus Google Scholar, J. D. Cell Res. 2004; PubMed Scopus Google Scholar), and to PMN during cell adhesion. a neutrophil elastase and cathepsin G for has been resulting in a of control of cathepsin G by this inhibitor elastase is present B. P. E. M. S. J. Cell Biol. 2005; PubMed Scopus Google Scholar). the of effect of to their to the membrane Indeed, releases proteases the of CD43 juxtamembranous cleavage The or of their the cell to this cleavage This also the by the cathepsin G inhibitor the of a RIP of CD43 in cancer cell lines (7Andersson C.X. Fernandez-Rodriguez J. Laos S. Baeckstrom D. Haass C. Hansson G.C. Biochem. J. 2005; 387: 377-384Crossref PubMed Scopus (39) Google Scholar), we investigated a cleavage of CD43 during neutrophil activation. The γ-secretase has been described in neutrophil J. L. J. L. K. N. 2002; PubMed Scopus (21) Google Scholar). A result from fusion with the γ-secretase recently been to neutrophil Jin 2006; PubMed Scopus Google Scholar), of the γ-secretase PMN activation by or by we a accumulation of the CD43-CTF, cells with three different γ-secretase This effect of γ-secretase was also on the CD43-CTF, resulting from PMN treatment by exogenous cathepsin G. The juxtamembranous by cathepsin G thus to CD43 susceptible to γ-secretase This role of cathepsin G is in it is by A role for in CD43 shedding had been CD43 shedding was inhibited by We here various metalloproteinase inhibitors, able to inhibit and/or membrane as of had effect on CD43 by for a was with the as E. Parent D. J. Immunol. 1994; 152: 3595-3605PubMed Google Scholar). thus to in CD43 proteolysis on We the of the role of the release of CD43 intracellular fragments, by this release with Our results γ-secretase no effect on neutrophil or inhibit neutrophil adhesion. Although this CD43, to membrane receptors processed by these results neutrophil adhesion the release of intracellular from shed transmembrane PMN activation by results in the shedding of of membrane The CD43 molecules with proteins and are in the and participate to cell P. M.R. Sanchez-Madrid F. Blood. 1995; PubMed Google Scholar, S. F. Halbwachs-Mecarelli L. Blood. PubMed Google Scholar). The proteolysis by of fragments, in the membrane after CD43 shedding, as the of by a R. 2004; 5: Scopus Google Scholar). with the E-cadherin cleavage by from the P. J. G. A. S. L. P. S. T. J. 2002; 21: PubMed Scopus Google Scholar), γ-secretase effect on CD43 the release of with CD43 the of with functions P.D. Cannon J.L. Bandukwala H.S. Blaine K.M. Schilling A.B. Swier K. Sperling A.I. Blood. 2007; 110: 2974-2982Crossref PubMed Scopus (21) Google Scholar, J. J.M. J. Immunol. Google Scholar) to this on CD43 molecules with Indeed, has been to both the and the intracellular domain of CD43 P.D. Cannon J.L. Bandukwala H.S. Blaine K.M. Schilling A.B. Swier K. Sperling A.I. Blood. 2007; 110: 2974-2982Crossref PubMed Scopus (21) Google Scholar). as a of by transmembrane molecules after the shedding of their extracellular R. 2004; 5: Scopus Google Scholar). processing also release CD43 intracellular with and properties. Indeed, a has been described in CD43 in cell lines, CD43 intracellular domain was to in the nucleus and an up-regulation of genes (9Andersson C.X. Fernandez-Rodriguez J. Laos S. Sikut R. Sikut A. Baeckstrom D. Hansson G.C. Biochem. Biophys. Res. Commun. 2004; 316: 12-17Crossref PubMed Scopus (24) Google Scholar). is to in neutrophils by which also CD43 CD43 intracytoplasmic these in the in neutrophils in the or of γ-secretase We here A is by to of CD43 shedding has been to in on PMN during Pulido R. M.R. Sanchez-Madrid F. 1991; Full Text PDF PubMed Scopus (67) Google Scholar) and during PMN to inflammation S. Halbwachs-Mecarelli L. P. Bessou G. M. Porteu F. Immunol. 1995; PubMed Scopus Google Scholar). Our data the juxtamembranous This cleavage was by Western blotting, of the of by the of CD43, by or in or by of exogenous cathepsin G, a of about 50% of the PMN CD43 This is of the 50% of CD43 in during PMN adhesion and S. Seveau S. Lesavre P. Robinson M.K. Halbwachs-Mecarelli L. Cell Adhes. Commun. 1998; 5: 151-160Crossref PubMed Scopus (24) Google Scholar) or in during PMN to S. Halbwachs-Mecarelli L. P. Bessou G. M. Porteu F. Immunol. 1995; PubMed Scopus Google Scholar). This an CD43 molecules on of to cathepsin G cleavage, to by membrane PMN activation in CD43 juxtamembranous cleavage by cathepsin G, while from the this release the extracellular domain of CD43, has been described in human and identified to the protein K. R. T. J. K. 1992; PubMed Scopus Google Scholar). We the of a function for CD43s, by with soluble PSGL-1, which has been to functions J. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar). soluble has been to inhibit the adhesion of colon cells to endothelial cells R. S. M. J. Biol. 1994; Full Text PDF PubMed Google Scholar). We CD43s, as an E-selectin ligand (10Fuhlbrigge R.C. King S.L. Sackstein R. Kupper T.S. Blood. 2006; 107: 1421-1426Crossref PubMed Scopus (95) Google Scholar, 11Matsumoto M. Atarashi K. Umemoto E. Furukawa Y. Shigeta A. Miyasaka M. Hirata T. J. Immunol. 2005; 175: 8042-8050Crossref PubMed Scopus (101) Google Scholar), with cell surface to leukocyte Our data this we the adhesion of neutrophils to under flow conditions, while it with their static adhesion. a with in where neutrophils to the a to result in CD43 shedding (2Nathan C. Xie Q.W. Halbwachs-Mecarelli L. Jin W.W. J. Cell Biol. 1993; 122: 243-256Crossref PubMed Scopus (115) Google Scholar, 16Lopez S. Seveau S. Lesavre P. Robinson M.K. Halbwachs-Mecarelli L. Cell Adhes. Commun. 1998; 5: 151-160Crossref PubMed Scopus (24) Google Scholar). The effect of is of the of leukocyte by recombinant a with a of and J. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar). CD43 shedding during neutrophil activation results in with adhesion The of PMN adhesion to by of participate to the of PMN for the resolution of as was for shedding (1Garton K.J. Gough P.J. Raines E.W. J. Leuk. Biol. 2006; 79: 1105-1116Crossref PubMed Scopus (191) Google Scholar). This has to to the of membrane receptors in responses S. M. 23: PubMed Google Scholar). cathepsin G together with to the shedding of various adhesion molecules or cytokine receptors and has been to with neutrophil by several in Immunol. 2006; PubMed Scopus Google Scholar). Our data a function for cathepsin G, CD43 which important for leukocyte in of CD43 to leukocyte and trafficking P.D. Cannon J.L. Bandukwala H.S. Blaine K.M. Schilling A.B. Swier K. Sperling A.I. Blood. 2007; 110: 2974-2982Crossref PubMed Scopus (21) Google Scholar, P. M.R. Sanchez-Madrid F. Blood. 1995; PubMed Google Scholar, S. F. Halbwachs-Mecarelli L. Blood. PubMed Google Scholar). We and from for metalloproteinase