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Proc Natl Acad Sci U S A. Aug 1, 1993; 90(15): 7158–7162.

Binding to heparan sulfate or heparin enhances neutrophil responses to interleukin 8.


The interaction of interleukin 8 (IL-8) with heparin was studied by using synthetic IL-8 analogs with C- and N-terminal truncations. Elimination of the N-terminal region preceding the first cysteine, which constitutes the IL-8 receptor binding site, did not affect the affinity to heparin-Sepharose. Affinity, however, decreased with progressive truncation at the C terminus, and no binding was observed when the C-terminal alpha-helix was eliminated. The effect of heparin and other glycosaminoglycans on IL-8 activity was also tested. When IL-8 was applied together with heparan sulfate, neutrophil chemotaxis in vitro was enhanced up to 4-fold, and the stimulus-dependent increase in cytosolic free Ca2+ increased markedly in both rate and peak value. Heparin had a similar effect on the Ca2+ response but did not enhance chemotaxis. The glycosaminoglycans by themselves did not elicit neutrophil responses. Their enhancing effect was restricted to stimulation with IL-8 and was not observed when the unrelated chemoattractant fMet-Ile-Phe-Leu was used as the stimulus. Elastase released from stimulated neutrophils was inhibited by heparin, heparan sulfate, and, to a lesser extent, chondroitin sulfate B, confirming previous observations. Taken together, these results suggest that heparan sulfate, which is present on the endothelial cell surface and in the basement membrane, may have a dual function in diapedesis, promotion of IL-8-dependent transmigration of neutrophils, and protection of the tissue microenvironment from damage by lytic enzymes released from the migrating cells.

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  • Oppenheim JJ, Zachariae CO, Mukaida N, Matsushima K. Properties of the novel proinflammatory supergene "intercrine" cytokine family. Annu Rev Immunol. 1991;9:617–648. [PubMed]
  • Sherry B, Cerami A. Small cytokine superfamily. Curr Opin Immunol. 1991 Feb;3(1):56–60. [PubMed]
  • Baggiolini M, Clark-Lewis I. Interleukin-8, a chemotactic and inflammatory cytokine. FEBS Lett. 1992 Jul 27;307(1):97–101. [PubMed]
  • Barber AJ, Käser-Glanzmann R, Jakábová M, Lüscher EF. Characterization of a chondroitin 4 -sulfate proteoglycan carrier for heparin neutralizing activity (platelet factor 4 ) released from human blood platelets. Biochim Biophys Acta. 1972 Dec 29;286(2):312–329. [PubMed]
  • Handin RI, Cohen HJ. Purification and binding properties of human platelet factor four. J Biol Chem. 1976 Jul 25;251(14):4273–4282. [PubMed]
  • Loscalzo J, Melnick B, Handin RI. The interaction of platelet factor four and glycosaminoglycans. Arch Biochem Biophys. 1985 Jul;240(1):446–455. [PubMed]
  • Zucker MB, Katz IR, Thorbecke GJ, Milot DC, Holt J. Immunoregulatory activity of peptides related to platelet factor 4. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7571–7574. [PMC free article] [PubMed]
  • Talpas CJ, Walz DA, Lee L. 1H-NMR studies of bovine platelet factor 4: histidine assignments and interactions with heparin. Biochim Biophys Acta. 1991 Jun 24;1078(2):208–218. [PubMed]
  • Clore GM, Appella E, Yamada M, Matsushima K, Gronenborn AM. Determination of the secondary structure of interleukin-8 by nuclear magnetic resonance spectroscopy. J Biol Chem. 1989 Nov 15;264(32):18907–18911. [PubMed]
  • Clore GM, Appella E, Yamada M, Matsushima K, Gronenborn AM. Three-dimensional structure of interleukin 8 in solution. Biochemistry. 1990 Feb 20;29(7):1689–1696. [PubMed]
  • Baldwin ET, Weber IT, St Charles R, Xuan JC, Appella E, Yamada M, Matsushima K, Edwards BF, Clore GM, Gronenborn AM, et al. Crystal structure of interleukin 8: symbiosis of NMR and crystallography. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):502–506. [PMC free article] [PubMed]
  • St Charles R, Walz DA, Edwards BF. The three-dimensional structure of bovine platelet factor 4 at 3.0-A resolution. J Biol Chem. 1989 Feb 5;264(4):2092–2099. [PubMed]
  • Hébert CA, Vitangcol RV, Baker JB. Scanning mutagenesis of interleukin-8 identifies a cluster of residues required for receptor binding. J Biol Chem. 1991 Oct 5;266(28):18989–18994. [PubMed]
  • Clark-Lewis I, Schumacher C, Baggiolini M, Moser B. Structure-activity relationships of interleukin-8 determined using chemically synthesized analogs. Critical role of NH2-terminal residues and evidence for uncoupling of neutrophil chemotaxis, exocytosis, and receptor binding activities. J Biol Chem. 1991 Dec 5;266(34):23128–23134. [PubMed]
  • Rot A, Henderson LE, Copeland TD, Leonard EJ. A series of six ligands for the human formyl peptide receptor: tetrapeptides with high chemotactic potency and efficacy. Proc Natl Acad Sci U S A. 1987 Nov;84(22):7967–7971. [PMC free article] [PubMed]
  • Eggleton P, Gargan R, Fisher D. Rapid method for the isolation of neutrophils in high yield without the use of dextran or density gradient polymers. J Immunol Methods. 1989 Jul 6;121(1):105–113. [PubMed]
  • Grynkiewicz G, Poenie M, Tsien RY. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed]
  • Kernen P, Wymann MP, von Tscharner V, Deranleau DA, Tai PC, Spry CJ, Dahinden CA, Baggiolini M. Shape changes, exocytosis, and cytosolic free calcium changes in stimulated human eosinophils. J Clin Invest. 1991 Jun;87(6):2012–2017. [PMC free article] [PubMed]
  • Dewald B, Baggiolini M. Evaluation of PAF antagonists using human neutrophils in a microtiter plate assay. Biochem Pharmacol. 1987 Aug 1;36(15):2505–2510. [PubMed]
  • Ferran DS, Sobel M, Harris RB. Design and synthesis of a helix heparin-binding peptide. Biochemistry. 1992 Jun 2;31(21):5010–5016. [PubMed]
  • Pratt CW, Church FC. Antithrombin: structure and function. Semin Hematol. 1991 Jan;28(1):3–9. [PubMed]
  • Eriksson AE, Cousens LS, Weaver LH, Matthews BW. Three-dimensional structure of human basic fibroblast growth factor. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3441–3445. [PMC free article] [PubMed]
  • Zhang JD, Cousens LS, Barr PJ, Sprang SR. Three-dimensional structure of human basic fibroblast growth factor, a structural homolog of interleukin 1 beta. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3446–3450. [PMC free article] [PubMed]
  • McCaffrey TA, Falcone DJ, Du B. Transforming growth factor-beta 1 is a heparin-binding protein: identification of putative heparin-binding regions and isolation of heparins with varying affinity for TGF-beta 1. J Cell Physiol. 1992 Aug;152(2):430–440. [PubMed]
  • von Tscharner V, Prod'hom B, Baggiolini M, Reuter H. Ion channels in human neutrophils activated by a rise in free cytosolic calcium concentration. Nature. 324(6095):369–372. [PubMed]
  • Kjellén L, Lindahl U. Proteoglycans: structures and interactions. Annu Rev Biochem. 1991;60:443–475. [PubMed]
  • Hardingham TE, Fosang AJ. Proteoglycans: many forms and many functions. FASEB J. 1992 Feb 1;6(3):861–870. [PubMed]
  • Gallagher JT, Lyon M, Steward WP. Structure and function of heparan sulphate proteoglycans. Biochem J. 1986 Jun 1;236(2):313–325. [PMC free article] [PubMed]
  • Rot A. Neutrophil attractant/activation protein-1 (interleukin-8) induces in vitro neutrophil migration by haptotactic mechanism. Eur J Immunol. 1993 Jan;23(1):303–306. [PubMed]
  • Nathan C, Sporn M. Cytokines in context. J Cell Biol. 1991 Jun;113(5):981–986. [PMC free article] [PubMed]
  • Burgess WH, Maciag T. The heparin-binding (fibroblast) growth factor family of proteins. Annu Rev Biochem. 1989;58:575–606. [PubMed]
  • Roberts R, Gallagher J, Spooncer E, Allen TD, Bloomfield F, Dexter TM. Heparan sulphate bound growth factors: a mechanism for stromal cell mediated haemopoiesis. Nature. 1988 Mar 24;332(6162):376–378. [PubMed]
  • Lantz M, Thysell H, Nilsson E, Olsson I. On the binding of tumor necrosis factor (TNF) to heparin and the release in vivo of the TNF-binding protein I by heparin. J Clin Invest. 1991 Dec;88(6):2026–2031. [PMC free article] [PubMed]
  • Lortat-Jacob H, Grimaud JA. Interferon-gamma C-terminal function: new working hypothesis. Heparan sulfate and heparin, new targets for IFN-gamma, protect, relax the cytokine and regulate its activity. Cell Mol Biol. 1991;37(3):253–260. [PubMed]
  • Ruoslahti E, Yamaguchi Y. Proteoglycans as modulators of growth factor activities. Cell. 1991 Mar 8;64(5):867–869. [PubMed]
  • Sudhalter J, Folkman J, Svahn CM, Bergendal K, D'Amore PA. Importance of size, sulfation, and anticoagulant activity in the potentiation of acidic fibroblast growth factor by heparin. J Biol Chem. 1989 Apr 25;264(12):6892–6897. [PubMed]
  • Saksela O, Moscatelli D, Sommer A, Rifkin DB. Endothelial cell-derived heparan sulfate binds basic fibroblast growth factor and protects it from proteolytic degradation. J Cell Biol. 1988 Aug;107(2):743–751. [PMC free article] [PubMed]
  • Yayon A, Klagsbrun M, Esko JD, Leder P, Ornitz DM. Cell surface, heparin-like molecules are required for binding of basic fibroblast growth factor to its high affinity receptor. Cell. 1991 Feb 22;64(4):841–848. [PubMed]
  • Rot A. Endothelial cell binding of NAP-1/IL-8: role in neutrophil emigration. Immunol Today. 1992 Aug;13(8):291–294. [PubMed]
  • Rot A. Binding of neutrophil attractant/activation protein-1 (interleukin 8) to resident dermal cells. Cytokine. 1992 Sep;4(5):347–352. [PubMed]
  • Huber AR, Kunkel SL, Todd RF, 3rd, Weiss SJ. Regulation of transendothelial neutrophil migration by endogenous interleukin-8. Science. 1991 Oct 4;254(5028):99–102. [PubMed]
  • Redini F, Tixier JM, Petitou M, Choay J, Robert L, Hornebeck W. Inhibition of leucocyte elastase by heparin and its derivatives. Biochem J. 1988 Jun 1;252(2):515–519. [PMC free article] [PubMed]
  • Avila JL, Convit J. Physicochemical characteristics of the glycosaminoglycan-lysosomal enzyme interaction in vitro. A model of control of leucocytic lysosomal activity. Biochem J. 1976 Nov 15;160(2):129–136. [PMC free article] [PubMed]

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