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J Clin Invest. Aug 1, 1997; 100(3): 522–529.
PMCID: PMC508218

Prevention of In vitro neutrophil adhesion to endothelial cells through shedding of L-selectin by C-reactive protein and peptides derived from C-reactive protein.


C-Reactive protein (CRP), the classic acute-phase reactant in humans, diminishes accumulation of neutrophils at inflammatory sites. To evaluate the underlying mechanisms, we have studied whether CRP and peptides derived from CRP could affect the first step of neutrophil extravasation, the L-selectin-mediated interaction of neutrophils with endothelial cells. CRP markedly attenuated attachment of human neutrophils to cultured LPS-activated human coronary artery and pulmonary microvascular endothelial cells with apparent IC50 values of 20 and 22 microg/ml, respectively. At similar concentrations, CRP rapidly downregulated the expression of L-selectin on the neutrophil surface, whereas it failed to affect expression of CD11b and CD45 or to induce granule enzyme release. The loss of L-selectin was due to cleavage and shedding of the molecule from the cell surface, as quantitated by the soluble form of L-selectin in cell-free supernatants. The effects of CRP could be prevented by an anti-CRP antiserum and by KD-IX-73-4, which inhibits shedding of L-selectin. Inhibition of adhesion with CRP was additive with function-blocking anti-E-selectin and anti-CD18 antibodies, but was not additive with anti-L-selectin antibody. Neutrophil attachment and L-selectin expression were also diminished by CRP peptides 174-185 and 201-206, but not peptide 77-82, albeit these peptides showed a weaker inhibitory effect than the parent protein. These studies indicate a specific activation-independent action of CRP and CRP peptides 174-185 and 201-206 on expression of L-selectin, and suggest that by attenuating neutrophil adhesion to the endothelium and consequently neutrophil traffic into tissues, native CRP and peptides 174-185 and 201-206 may be major mechanisms to attenuate or limit the inflammatory response.

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Selected References

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  • Pepys MB. C-reactive protein fifty years on. Lancet. 1981 Mar 21;1(8221):653–657. [PubMed]
  • Kushner I. The acute phase response: an overview. Methods Enzymol. 1988;163:373–383. [PubMed]
  • Müller H, Fehr J. Binding of C-reactive protein to human polymorphonuclear leukocytes: evidence for association of binding sites with Fc receptors. J Immunol. 1986 Mar 15;136(6):2202–2207. [PubMed]
  • Buchta R, Pontet M, Fridkin M. Binding of C-reactive protein to human neutrophils. FEBS Lett. 1987 Jan 26;211(2):165–168. [PubMed]
  • Kew RR, Hyers TM, Webster RO. Human C-reactive protein inhibits neutrophil chemotaxis in vitro: possible implications for the adult respiratory distress syndrome. J Lab Clin Med. 1990 Mar;115(3):339–345. [PubMed]
  • Heuertz RM, Piquette CA, Webster RO. Rabbits with elevated serum C-reactive protein exhibit diminished neutrophil infiltration and vascular permeability in C5a-induced alveolitis. Am J Pathol. 1993 Jan;142(1):319–328. [PMC free article] [PubMed]
  • Ahmed N, Thorley R, Xia D, Samols D, Webster RO. Transgenic mice expressing rabbit C-reactive protein exhibit diminished chemotactic factor-induced alveolitis. Am J Respir Crit Care Med. 1996 Mar;153(3):1141–1147. [PubMed]
  • Filep J, Földes-Filep E. Effects of C-reactive protein on human neutrophil granulocytes challenged with N-formyl-methionyl-leucyl-phenylalanine and platelet-activating factor. Life Sci. 1989;44(8):517–524. [PubMed]
  • Shephard EG, Beer SM, Anderson R, Strachan AF, Nel AE, de Beer FC. Generation of biologically active C-reactive protein peptides by a neutral protease on the membrane of phorbol myristate acetate-stimulated neutrophils. J Immunol. 1989 Nov 1;143(9):2974–2981. [PubMed]
  • Shephard EG, Anderson R, Rosen O, Myer MS, Fridkin M, Strachan AF, De Beer FC. Peptides generated from C-reactive protein by a neutrophil membrane protease. Amino acid sequence and effects of peptides on neutrophil oxidative metabolism and chemotaxis. J Immunol. 1990 Sep 1;145(5):1469–1476. [PubMed]
  • Heuertz RM, Ahmed N, Webster RO. Peptides derived from C-reactive protein inhibit neutrophil alveolitis. J Immunol. 1996 May 1;156(9):3412–3417. [PubMed]
  • Butcher EC. Leukocyte-endothelial cell recognition: three (or more) steps to specificity and diversity. Cell. 1991 Dec 20;67(6):1033–1036. [PubMed]
  • Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994 Jan 28;76(2):301–314. [PubMed]
  • Lawrence MB, Springer TA. Leukocytes roll on a selectin at physiologic flow rates: distinction from and prerequisite for adhesion through integrins. Cell. 1991 May 31;65(5):859–873. [PubMed]
  • Ley K, Gaehtgens P, Fennie C, Singer MS, Lasky LA, Rosen SD. Lectin-like cell adhesion molecule 1 mediates leukocyte rolling in mesenteric venules in vivo. Blood. 1991 Jun 15;77(12):2553–2555. [PubMed]
  • Kishimoto TK, Jutila MA, Berg EL, Butcher EC. Neutrophil Mac-1 and MEL-14 adhesion proteins inversely regulated by chemotactic factors. Science. 1989 Sep 15;245(4923):1238–1241. [PubMed]
  • Jutila MA, Rott L, Berg EL, Butcher EC. Function and regulation of the neutrophil MEL-14 antigen in vivo: comparison with LFA-1 and MAC-1. J Immunol. 1989 Nov 15;143(10):3318–3324. [PubMed]
  • Kishimoto TK, Warnock RA, Jutila MA, Butcher EC, Lane C, Anderson DC, Smith CW. Antibodies against human neutrophil LECAM-1 (LAM-1/Leu-8/DREG-56 antigen) and endothelial cell ELAM-1 inhibit a common CD18-independent adhesion pathway in vitro. Blood. 1991 Aug 1;78(3):805–811. [PubMed]
  • Leeuwenberg JF, von Asmuth EJ, Jeunhomme TM, Buurman WA. IFN-gamma regulates the expression of the adhesion molecule ELAM-1 and IL-6 production by human endothelial cells in vitro. J Immunol. 1990 Oct 1;145(7):2110–2114. [PubMed]
  • Walcheck B, Kahn J, Fisher JM, Wang BB, Fisk RS, Payan DG, Feehan C, Betageri R, Darlak K, Spatola AF, et al. Neutrophil rolling altered by inhibition of L-selectin shedding in vitro. Nature. 1996 Apr 25;380(6576):720–723. [PubMed]
  • Dewald B, Bretz U, Baggiolini M. Release of gelatinase from a novel secretory compartment of human neutrophils. J Clin Invest. 1982 Sep;70(3):518–525. [PMC free article] [PubMed]
  • Spertini O, Luscinskas FW, Kansas GS, Munro JM, Griffin JD, Gimbrone MA, Jr, Tedder TF. Leukocyte adhesion molecule-1 (LAM-1, L-selectin) interacts with an inducible endothelial cell ligand to support leukocyte adhesion. J Immunol. 1991 Oct 15;147(8):2565–2573. [PubMed]
  • Spertini O, Luscinskas FW, Gimbrone MA, Jr, Tedder TF. Monocyte attachment to activated human vascular endothelium in vitro is mediated by leukocyte adhesion molecule-1 (L-selectin) under nonstatic conditions. J Exp Med. 1992 Jun 1;175(6):1789–1792. [PMC free article] [PubMed]
  • Díaz-González F, González-Alvaro I, Campanero MR, Mollinedo F, del Pozo MA, Muñoz C, Pivel JP, Sánchez-Madrid F. Prevention of in vitro neutrophil-endothelial attachment through shedding of L-selectin by nonsteroidal antiinflammatory drugs. J Clin Invest. 1995 Apr;95(4):1756–1765. [PMC free article] [PubMed]
  • Robey FA, Ohura K, Futaki S, Fujii N, Yajima H, Goldman N, Jones KD, Wahl S. Proteolysis of human C-reactive protein produces peptides with potent immunomodulating activity. J Biol Chem. 1987 May 25;262(15):7053–7057. [PubMed]
  • Shephard EG, Anderson R, Rosen O, Fridkin M. C-reactive protein (CRP) peptides inactivate enolase in human neutrophils leading to depletion of intracellular ATP and inhibition of superoxide generation. Immunology. 1992 May;76(1):79–85. [PMC free article] [PubMed]
  • Goetzl EJ, Austen KF. Structural determinants of the eosinophil: chemotactic activity of the acidic tetrapeptides of eosinophil chemotactic factor of anaphylaxis. J Exp Med. 1976 Dec 1;144(6):1424–1437. [PMC free article] [PubMed]
  • Barna BP, Eppstein DA, Thomassen MJ, Nestor JJ, Jr, Ho T, Medendorp SV, Deodhar SD. Therapeutic effects of a synthetic peptide of C-reactive protein in pre-clinical tumor models. Cancer Immunol Immunother. 1993;36(3):171–176. [PubMed]
  • Barna BP, Thomassen MJ, Zhou P, Pettay J, Singh-Burgess S, Deodhar SD. Activation of alveolar macrophage TNF and MCP-1 expression in vivo by a synthetic peptide of C-reactive protein. J Leukoc Biol. 1996 Mar;59(3):397–402. [PubMed]
  • Berg M, James SP. Human neutrophils release the Leu-8 lymph node homing receptor during cell activation. Blood. 1990 Dec 1;76(11):2381–2388. [PubMed]
  • Miller LJ, Bainton DF, Borregaard N, Springer TA. Stimulated mobilization of monocyte Mac-1 and p150,95 adhesion proteins from an intracellular vesicular compartment to the cell surface. J Clin Invest. 1987 Aug;80(2):535–544. [PMC free article] [PubMed]
  • Kahn J, Ingraham RH, Shirley F, Migaki GI, Kishimoto TK. Membrane proximal cleavage of L-selectin: identification of the cleavage site and a 6-kD transmembrane peptide fragment of L-selectin. J Cell Biol. 1994 Apr;125(2):461–470. [PMC free article] [PubMed]
  • Tedder TF, Steeber DA, Chen A, Engel P. The selectins: vascular adhesion molecules. FASEB J. 1995 Jul;9(10):866–873. [PubMed]
  • Palecanda A, Walcheck B, Bishop DK, Jutila MA. Rapid activation-independent shedding of leukocyte L-selectin induced by cross-linking of the surface antigen. Eur J Immunol. 1992 May;22(5):1279–1286. [PubMed]
  • Migaki GI, Kahn J, Kishimoto TK. Mutational analysis of the membrane-proximal cleavage site of L-selectin: relaxed sequence specificity surrounding the cleavage site. J Exp Med. 1995 Aug 1;182(2):549–557. [PMC free article] [PubMed]
  • Chen A, Engel P, Tedder TF. Structural requirements regulate endoproteolytic release of the L-selectin (CD62L) adhesion receptor from the cell surface of leukocytes. J Exp Med. 1995 Aug 1;182(2):519–530. [PMC free article] [PubMed]
  • Du Clos TW, Mold C, Paterson PY, Alroy J, Gewurz H. Localization of C-reactive protein in inflammatory lesions of experimental allergic encephalomyelitis. Clin Exp Immunol. 1981 Mar;43(3):565–573. [PMC free article] [PubMed]
  • Vigushin DM, Pepys MB, Hawkins PN. Metabolic and scintigraphic studies of radioiodinated human C-reactive protein in health and disease. J Clin Invest. 1993 Apr;91(4):1351–1357. [PMC free article] [PubMed]
  • Shields MJ. A hypothesis resolving the apparently disparate activities of native and altered forms of human C-reactive protein. Immunol Res. 1993;12(1):37–47. [PubMed]

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