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. 1988 May;81(5):1310–1316. doi: 10.1172/JCI113456

Fibronectin degradation products containing the cytoadhesive tetrapeptide stimulate human neutrophil degranulation.

Y T Wachtfogel 1, W Abrams 1, U Kucich 1, G Weinbaum 1, M Schapira 1, R W Colman 1
PMCID: PMC442557  PMID: 2966812

Abstract

We investigated whether adhesive glycoproteins, such as fibronectin or fibrinogen, could function to provide a nidus for neutrophil degranulation. Elastase release in recalcified plasma was normal in afibrinogenemic plasma, but 73% less in plasma depleted of fibronectin. Proteolytic digests of fibronectin, but not intact fibronectin (50-1,000 micrograms/ml), induced a concentration-dependent release of neutrophil elastase and lactoferrin. MAbs N293, which recognized the mid-molecule of fibronectin, N294, which was directed toward the 11-kD cell adhesive fragment, and N295, generated against the amino terminal of the 11-kD fragment, inhibited the release of elastase by 7, 24, and 60%, respectively. The cytoadhesive tetrapeptide portion of fibronectin, Arg-Gly-Asp-Ser (250-1,000 micrograms/ml), released 1.94 +/- 0.10 micrograms/ml of elastase from 10(7) neutrophils, in contrast to the lack of release by the control hexapeptide, Arg-Gly-Tyr-Ser-Leu-Gly. Plasmin appeared to be the enzyme responsible for fibronectin cleavage, since neutrophil elastase release in plasma that had been depleted of plasminogen was decreased and reconstitution of plasminogen-deficient plasma with purified plasminogen corrected the abnormal release. Plasmin cleaved fibronectin to multiple degradation products, each less than 200 kD. This fibronectin digest released 1.05 microgram/ml of elastase from 10(7) neutrophils. We suggest that the activation of plasminogen leads to the formation of fibronectin degradation products capable of functioning as agonists for neutrophils.

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Selected References

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  1. Alexander J. W., McClellan M. A., Ogle C. K., Ogle J. D. Consumptive opsoninopathy: possible pathogenesis in lethal and opportunistic infections. Ann Surg. 1976 Dec;184(6):672–678. doi: 10.1097/00000658-197612000-00002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Becker E. L. Some interrelations of neutrophil chemotaxis, lysosomal enzyme secretion, and phagocytosis as revealed by synthetic peptides. Am J Pathol. 1976 Nov;85(2):385–394. [PMC free article] [PubMed] [Google Scholar]
  3. Bretz U., Baggiolini M. Biochemical and morphological characterization of azurophil and specific granules of human neutrophilic polymorphonuclear leukocytes. J Cell Biol. 1974 Oct;63(1):251–269. doi: 10.1083/jcb.63.1.251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brower M. S., Levin R. I., Garry K. Human neutrophil elastase modulates platelet function by limited proteolysis of membrane glycoproteins. J Clin Invest. 1985 Feb;75(2):657–666. doi: 10.1172/JCI111744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brower M. S., Walz D. A., Garry K. E., Fenton J. W., 2nd Human neutrophil elastase alters human alpha-thrombin function: limited proteolysis near the gamma-cleavage site results in decreased fibrinogen clotting and platelet-stimulatory activity. Blood. 1987 Mar;69(3):813–819. [PubMed] [Google Scholar]
  6. Collen D. Identification and some properties of a new fast-reacting plasmin inhibitor in human plasma. Eur J Biochem. 1976 Oct 1;69(1):209–216. doi: 10.1111/j.1432-1033.1976.tb10875.x. [DOI] [PubMed] [Google Scholar]
  7. Cosio F. G., Bakaletz A. P. Abnormal plasma fibronectin levels in patients with proteinuria. J Lab Clin Med. 1984 Dec;104(6):867–872. [PubMed] [Google Scholar]
  8. Craddock P. R., Hammerschmidt D., White J. G., Dalmosso A. P., Jacob H. S. Complement (C5-a)-induced granulocyte aggregation in vitro. A possible mechanism of complement-mediated leukostasis and leukopenia. J Clin Invest. 1977 Jul;60(1):260–264. doi: 10.1172/JCI108763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Deutsch D. G., Mertz E. T. Plasminogen: purification from human plasma by affinity chromatography. Science. 1970 Dec 4;170(3962):1095–1096. doi: 10.1126/science.170.3962.1095. [DOI] [PubMed] [Google Scholar]
  10. Doolittle R. F., Watt K. W., Cottrell B. A., Strong D. D., Riley M. The amino acid sequence of the alpha-chain of human fibrinogen. Nature. 1979 Aug 9;280(5722):464–468. doi: 10.1038/280464a0. [DOI] [PubMed] [Google Scholar]
  11. Fletcher M. P., Gallin J. I. Degranulating stimuli increase the availability of receptors on human neutrophils for the chemoattractant f-met-leu-phe. J Immunol. 1980 Apr;124(4):1585–1588. [PubMed] [Google Scholar]
  12. Fraker P. J., Speck J. C., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
  13. Gardner J. M., Hynes R. O. Interaction of fibronectin with its receptor on platelets. Cell. 1985 Sep;42(2):439–448. doi: 10.1016/0092-8674(85)90101-1. [DOI] [PubMed] [Google Scholar]
  14. Gartner T. K., Bennett J. S. The tetrapeptide analogue of the cell attachment site of fibronectin inhibits platelet aggregation and fibrinogen binding to activated platelets. J Biol Chem. 1985 Oct 5;260(22):11891–11894. [PubMed] [Google Scholar]
  15. Ginsberg M., Pierschbacher M. D., Ruoslahti E., Marguerie G., Plow E. Inhibition of fibronectin binding to platelets by proteolytic fragments and synthetic peptides which support fibroblast adhesion. J Biol Chem. 1985 Apr 10;260(7):3931–3936. [PubMed] [Google Scholar]
  16. Goetzl E. J., Austen K. F. Stimulation of human neutrophil leukocyte aerobic glucose metabolism by purified chemotactic factors. J Clin Invest. 1974 Feb;53(2):591–599. doi: 10.1172/JCI107594. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Goldstein I. M., Weissmann G. Generation of C5-derived lysosomal enzyme-releasing activity (C5a) by lysates of leukocyte lysosomes. J Immunol. 1974 Nov;113(5):1583–1588. [PubMed] [Google Scholar]
  18. Goodnough L. T., Saito H., Ratnoff O. D. Fibronectin levels in congenital thrombocytopenia: Schulman's syndrome. N Engl J Med. 1982 Apr 15;306(15):938–939. doi: 10.1056/NEJM198204153061516. [DOI] [PubMed] [Google Scholar]
  19. HIRSCH J. G., COHN Z. A. Degranulation of polymorphonuclear leucocytes following phagocytosis of microorganisms. J Exp Med. 1960 Dec 1;112:1005–1014. doi: 10.1084/jem.112.6.1005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Harpel P. C., Mosesson M. W. Degradation of human fibrinogen by plasms alpha2-macroglobulin-enzyme complexes. J Clin Invest. 1973 Sep;52(9):2175–2184. doi: 10.1172/JCI107402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hawiger J., Timmons S., Kloczewiak M., Strong D. D., Doolittle R. F. gamma and alpha chains of human fibrinogen possess sites reactive with human platelet receptors. Proc Natl Acad Sci U S A. 1982 Mar;79(6):2068–2071. doi: 10.1073/pnas.79.6.2068. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Heiple J. M., Ossowski L. Human neutrophil plasminogen activator is localized in specific granules and is translocated to the cell surface by exocytosis. J Exp Med. 1986 Sep 1;164(3):826–840. doi: 10.1084/jem.164.3.826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hummer L., Munkner T., Sørensen S. S., Brandt N. J., Jacobsen B. B. Nation-wide TSH-screening with low recall rate. Laboratory results of a two-year study. Scand J Clin Lab Invest. 1982 Feb;42(1):49–55. doi: 10.1080/00365518209168049. [DOI] [PubMed] [Google Scholar]
  24. Humphries M. J., Olden K., Yamada K. M. A synthetic peptide from fibronectin inhibits experimental metastasis of murine melanoma cells. Science. 1986 Jul 25;233(4762):467–470. doi: 10.1126/science.3726541. [DOI] [PubMed] [Google Scholar]
  25. Hörmann H., Richter H., Jelinić V. Fibrinmonomer binding to macrophages mediated by fibrin-binding fibronectin fragments. Thromb Res. 1985 Jul 15;39(2):183–194. doi: 10.1016/0049-3848(85)90106-9. [DOI] [PubMed] [Google Scholar]
  26. Janoff A., Sloan B., Weinbaum G., Damiano V., Sandhaus R. A., Elias J., Kimbel P. Experimental emphysema induced with purified human neutrophil elastase: tissue localization of the instilled protease. Am Rev Respir Dis. 1977 Mar;115(3):461–478. doi: 10.1164/arrd.1977.115.3.461. [DOI] [PubMed] [Google Scholar]
  27. Jilek F., Hörmann H. Cold-insoluble globulin, II. Plasminolysis of cold-insoluble globulin. Hoppe Seylers Z Physiol Chem. 1977 Jan;358(1):133–136. [PubMed] [Google Scholar]
  28. Kaplan A. P., Kay A. B., Austen K. F. A prealbumin activator of prekallikrein. 3. Appearance of chemotactic activity for human neutrophils by the conversion of human prekallikrein to kallikrein. J Exp Med. 1972 Jan;135(1):81–97. doi: 10.1084/jem.135.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Keren G., Gordon P. A., Lee S. F., Stewart M., Gelfand E. T. Alterations in plasma-, monocyte-, and lymphocyte-associated fibronectin during cardiopulmonary bypass surgery. Am J Clin Pathol. 1985 May;83(5):629–633. doi: 10.1093/ajcp/83.5.629. [DOI] [PubMed] [Google Scholar]
  30. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  31. LeRoy E. C., Ager A., Gordon J. L. Effects of neutrophil elastase and other proteases on porcine aortic endothelial prostaglandin I2 production, adenine nucleotide release, and responses to vasoactive agents. J Clin Invest. 1984 Sep;74(3):1003–1010. doi: 10.1172/JCI111467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Marguerie G. A., Plow E. F., Edgington T. S. Human platelets possess an inducible and saturable receptor specific for fibrinogen. J Biol Chem. 1979 Jun 25;254(12):5357–5363. [PubMed] [Google Scholar]
  33. Matsuda M., Yamanaka T., Matsuda A. Distribution of fibronectin in plasma and liver in liver diseases. Clin Chim Acta. 1982 Feb 5;118(2-3):191–199. doi: 10.1016/0009-8981(82)90006-7. [DOI] [PubMed] [Google Scholar]
  34. McDonald J. A., Kelley D. G. Degradation of fibronectin by human leukocyte elastase. Release of biologically active fragments. J Biol Chem. 1980 Sep 25;255(18):8848–8858. [PubMed] [Google Scholar]
  35. McGuire W. W., Spragg R. G., Cohen A. B., Cochrane C. G. Studies on the pathogenesis of the adult respiratory distress syndrome. J Clin Invest. 1982 Mar;69(3):543–553. doi: 10.1172/JCI110480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Miekka S. I., Ingham K. C., Menache D. Rapid methods for isolation of human plasma fibronectin. Thromb Res. 1982 Jul 1;27(1):1–14. doi: 10.1016/0049-3848(82)90272-9. [DOI] [PubMed] [Google Scholar]
  37. Mosher D. F., Williams E. M. Fibronectin concentration is decreased in plasma of severely ill patients with disseminated intravascular coagulation. J Lab Clin Med. 1978 May;91(5):729–735. [PubMed] [Google Scholar]
  38. Naccache P. H., Showell H. J., Becker E. L., Sha'afi R. I. Transport of sodium, potassium, and calcium across rabbit polymorphonuclear leukocyte membranes. Effect of chemotactic factor. J Cell Biol. 1977 May;73(2):428–444. doi: 10.1083/jcb.73.2.428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Nakajima K., Powers J. C., Ashe B. M., Zimmerman M. Mapping the extended substrate binding site of cathepsin G and human leukocyte elastase. Studies with peptide substrates related to the alpha 1-protease inhibitor reactive site. J Biol Chem. 1979 May 25;254(10):4027–4032. [PubMed] [Google Scholar]
  40. Norris D. A., Clark R. A., Swigart L. M., Huff J. C., Weston W. L., Howell S. E. Fibronectin fragment(s) are chemotactic for human peripheral blood monocytes. J Immunol. 1982 Oct;129(4):1612–1618. [PubMed] [Google Scholar]
  41. O'Flaherty J. T., Ward P. A. Leukocyte aggregation induced by chemotactic factors: a review. Inflammation. 1978 Jun;3(2):177–194. doi: 10.1007/BF00910738. [DOI] [PubMed] [Google Scholar]
  42. Pierschbacher M. D., Ruoslahti E. Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule. Nature. 1984 May 3;309(5963):30–33. doi: 10.1038/309030a0. [DOI] [PubMed] [Google Scholar]
  43. Plow E. F., Freaney D. E., Plescia J., Miles L. A. The plasminogen system and cell surfaces: evidence for plasminogen and urokinase receptors on the same cell type. J Cell Biol. 1986 Dec;103(6 Pt 1):2411–2420. doi: 10.1083/jcb.103.6.2411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Plow E. F., Gramse M., Havemann K. Immunochemical discrimination of leukocyte elastase from plasmic degradation products of fibrinogen. J Lab Clin Med. 1983 Dec;102(6):858–869. [PubMed] [Google Scholar]
  45. Plow E. F. Leukocyte elastase release during blood coagulation. A potential mechanism for activation of the alternative fibrinolytic pathway. J Clin Invest. 1982 Mar;69(3):564–572. doi: 10.1172/JCI110482. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Proctor R. A., Prendergast E., Mosher D. F. Fibronectin mediates attachment of Staphylococcus aureus to human neutrophils. Blood. 1982 Apr;59(4):681–687. [PubMed] [Google Scholar]
  47. Pytela R., Pierschbacher M. D., Ginsberg M. H., Plow E. F., Ruoslahti E. Platelet membrane glycoprotein IIb/IIIa: member of a family of Arg-Gly-Asp--specific adhesion receptors. Science. 1986 Mar 28;231(4745):1559–1562. doi: 10.1126/science.2420006. [DOI] [PubMed] [Google Scholar]
  48. Rourke F. J., Blumenstock F. A., Kaplan J. E. Effect of fibronectin fragments on macrophage phagocytosis of gelatinized particles. J Immunol. 1984 Apr;132(4):1931–1936. [PubMed] [Google Scholar]
  49. Rubli E., Büssard S., Frei E., Lundsgaard-Hansen P., Pappova E. Plasma fibronectin and associated variables in surgical intensive care patients. Ann Surg. 1983 Mar;197(3):310–317. doi: 10.1097/00000658-198303000-00011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Ruggeri Z. M., Bader R., de Marco L. Glanzmann thrombasthenia: deficient binding of von Willebrand factor to thrombin-stimulated platelets. Proc Natl Acad Sci U S A. 1982 Oct;79(19):6038–6041. doi: 10.1073/pnas.79.19.6038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Schapira M., Despland E., Scott C. F., Boxer L. A., Colman R. W. Purified human plasma kallikrein aggregates human blood neutrophils. J Clin Invest. 1982 May;69(5):1199–1202. doi: 10.1172/JCI110557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Scott R. L., Sohmer P. R., MacDonald M. G. The effect of starvation and repletion on plasma fibronectin in man. JAMA. 1982 Oct 22;248(16):2025–2027. [PubMed] [Google Scholar]
  53. Shirakami A., Shigekiyo T., Hirai Y., Takeichi T., Kawauchi S., Saito S., Miyoshi K. Plasma fibronectin deficiency in eight members of one family. Lancet. 1986 Mar 1;1(8479):473–474. doi: 10.1016/s0140-6736(86)92931-4. [DOI] [PubMed] [Google Scholar]
  54. Spitznagel J. K., Dalldorf F. G., Leffell M. S., Folds J. D., Welsh I. R., Cooney M. H., Martin L. E. Character of azurophil and specific granules purified from human polymorphonuclear leukocytes. Lab Invest. 1974 Jun;30(6):774–785. [PubMed] [Google Scholar]
  55. ULMER D. D., VALLEE B. L., WACKER W. E. Metalloenzymes and myocardial infarction. II. Malic and lactic dehydrogenase activities and zinc concentrations in serum. N Engl J Med. 1956 Sep 6;255(10):450–456. doi: 10.1056/NEJM195609062551001. [DOI] [PubMed] [Google Scholar]
  56. Wachtfogel Y. T., Kucich U., James H. L., Scott C. F., Schapira M., Zimmerman M., Cohen A. B., Colman R. W. Human plasma kallikrein releases neutrophil elastase during blood coagulation. J Clin Invest. 1983 Nov;72(5):1672–1677. doi: 10.1172/JCI111126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Wachtfogel Y. T., Pixley R. A., Kucich U., Abrams W., Weinbaum G., Schapira M., Colman R. W. Purified plasma factor XIIa aggregates human neutrophils and causes degranulation. Blood. 1986 Jun;67(6):1731–1737. [PubMed] [Google Scholar]
  58. Wicki A. N., Clemetson K. J. Structure and function of platelet membrane glycoproteins Ib and V. Effects of leukocyte elastase and other proteases on platelets response to von Willebrand factor and thrombin. Eur J Biochem. 1985 Nov 15;153(1):1–11. doi: 10.1111/j.1432-1033.1985.tb09259.x. [DOI] [PubMed] [Google Scholar]
  59. Yoder M. C., Douglas S. D., Gerdes J., Kline J., Polin R. A. Plasma fibronectin in healthy newborn infants: respiratory distress syndrome and perinatal asphyxia. J Pediatr. 1983 May;102(5):777–780. doi: 10.1016/s0022-3476(83)80257-1. [DOI] [PubMed] [Google Scholar]

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