Download PDF
Thromb Haemost 1984; 52(03): 243-249
DOI: 10.1055/s-0038-1661188
Original Article
Schattauer GmbH Stuttgart

Plasminogen Activator and Plasminogen Activator Inhibitor Associated with Granulomatous Inflammation: A Study with Murine Leprosy

S Izaki
The Department of Dermatology, Iwate Medical University School of Medicine, Morloka, Iwate, Japan
,
T Hibino
The Department of Dermatology, Iwate Medical University School of Medicine, Morloka, Iwate, Japan
,
Y Isozaki
The Department of Dermatology, Iwate Medical University School of Medicine, Morloka, Iwate, Japan
,
P S Hsu
The Department of Dermatology, Iwate Medical University School of Medicine, Morloka, Iwate, Japan
,
M Izaki
The Department of Dermatology, Iwate Medical University School of Medicine, Morloka, Iwate, Japan
,
O Matsuo
*   The Department of Physiology, Kinki University School of Medicine, Sayama, Osaka, Japan
› Author Affiliations
Further Information

Publication History

Received 02 December 1983

Accepted 08 August 1984

Publication Date:
19 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

Plasminogen activator that is associated with the development of hypersensitivity granulomas (gPA) was partially purified from a saline soluble fraction of murine lepromas elicited in “resistant” mice, C57BL/6N. The gPA was shown to consist of two subspecies (23,000 and 48,000 in molecular weight) with essentially identical enzymologic properties. The gPA was found to be a relatively heat stable weakly alkaline serine proteinase with trypsin-like characteristics in the specificity for synthetic substrates and proteinase inhibitors. It showed a high affinity for H- D-Ile-Pro-Arg-pNA (Km = 1.4 × 10-4 M) H-D-Val-Leu-Lys- pNA (Km = 5.2 × 10-4 M), and L-pyroGlu-Gly-Arg-pNA (Km = 9.3 × 10-4 M). The gPA did not demonstrate antigenic cross reaction with urokinase-type or tissue-type plasminogen activator.

Two distinct enzymatic regulators of the gPA were also demonstrated in the saline soluble fraction of the hypersensitivity granulomas. The gPA and its regulation are assumed to be correlated with macrophage activation in the hypersensitivity granulomas

Key words

Plasminogen activator - Plasminogen activator inhibitor - Inflammation - Hypersensitivity granulomas - Murine leprosy
 

  • References

  • 1 Epstein WL, Krasnobrod H. The origin of epithelioid cells in experimental granulomas of man. Lab Invest 1968; 18: 190-195
    PubMedGoogle Scholar
  • 2 Black M, Epstein WL. Formation of multinucleate giant cells in organized epithelioid cell granulomas. Am J Pathol 1974; 74: 263-274
    PubMedGoogle Scholar
  • 3 Hsu WY, Hsu HF, Lust GL, Davis JR. Organized epithelioid cell granulomata elicited by schistosome eggs in experiemental animals. J Reticuloendothel Soc 1972; 12: 418-435
    PubMedGoogle Scholar
  • 4 von Lichtenberg F, Erickson DG, Sadun EH. Comparative histopathology of schistosome granulomas in the hamster. Am J Pathol 1973; 72: 149-168
    PubMedGoogle Scholar
  • 5 Epstein WL, Fukuyama K, Danno K, Kwan-Wong E. Granulomatous inflammation in normal and athymic mice infected with Schistosoma mansoni: an ultrastructural study. J Pathol 1979; 127: 207-215
    CrossrefPubMedGoogle Scholar
  • 6 Williams WJ, Erasmus DA, Jenkins D, Valerie-James EM, Davies T. A comparative study of the ultrastructure and histochemistry of sarcoid and tuberculous granulomas. In: Fifth International Conference on Sarcoidosis. Leninsky L, Macholda F. (Eds) Universita Karlova; Praha: 1971. pp 115-120
    Google Scholar
  • 7 Closs O, Haugen OA. Experimental murine leprosy 4. The gross appearance and microscopic features of the local infiltrate after subcutaneous inoculation of C3H and C57/BL mice with Mycobacterium lepraemurium. Acta Pathol Microbiol Scand [A] 1975; 83: 59-68
    PubMedGoogle Scholar
  • 8 Izaki S, Goldstein SM, Fukuyama K, Epstein WL. Fibrin deposition and clearance in chronic granulomatous inflammation: Correlation with T-cell function and proteinase inhibitor activity in tissue. J Invest Dermatol 1979; 73: 561-565
    CrossrefPubMedGoogle Scholar
  • 9 Izaki S, Isozaki Y, Satoh M, Hibino T, Kon S, Izaki M. Comparative study with two polar types of murine leprosy: An involvement of plasminogen activator and its possible regulating factor in the granulomatous tissue reaction. J Invest Dermatol 1983; 80: 81-85
    CrossrefPubMedGoogle Scholar
  • 10 Satoh M, Izaki S, Kon S, Izaki M. Fibrin deposition and fibroblasts proliferation in granuloma of murine leprosy. Comparison of two mouse strains with different immune reactions Experientia 1983; 39: 899-902
    CrossrefPubMedGoogle Scholar
  • 11 Kawaguchi Y, Matsuoka M. Observation of host reactions to murine leprosy bacilli in spread subcutaneous tissue preparations of various strains of mice. Jpn J Exp Med 1977; 47: 71-79
    PubMedGoogle Scholar
  • 12 Haugen OA, Skjorten F, Closs O. Experimental murine leprosy 8. Ultrastructural features of inflammatory exudate and bacterial morphology in C3H and C57BL mice after footpad inoculation with Mycobacterium lepraemurium. Acta Pathol Microbiol Scand [A] 1975; 83: 693-703
    PubMedGoogle Scholar
  • 13 Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193: 265-275
    PubMedGoogle Scholar
  • 14 Okamoto U, Nagamatsu Y, Tsuda Y, Okada Y. A new polypeptide substrate Suc-Tyr-Leu-Val-pNA, specific for spleen fibrinolytic proteinase (SFP). Biochem Biophys Res Commun 1980; 97: 28-32
    CrossrefPubMedGoogle Scholar
  • 15 Hibino T, Izaki S, Izaki M. Detection of serine proteinase inhibitors in human comified cells. Biochem Biophys Res Commun 1981; 101: 948-955
    CrossrefPubMedGoogle Scholar
  • 16 Segel IH. Enzyme kinetics: behaviour and analysis of rapid equilibrium and steady state enzyme systems. A Wiley-Interscience Publication; New York: 1975. pp 46-51
    Google Scholar
  • 17 Unkeless JC, Tobia A, Ossowski L, Quigley JP, Rifkin DB, Reich E. An enzymatic function associated with transformation of fibroblasts by oncogenic viruses. I. Chick embryo fibroblast cultures transformed by avian RNA tumor viruses. J Exp Med 1973; 137: 85-111
    CrossrefPubMedGoogle Scholar
  • 18 Deutsch DG, Mertz ET. Plasminogen; purification from human plasma by affinity chromatography. Science 1970; 170: 1095-1096
    CrossrefPubMedGoogle Scholar
  • 19 Chase Jr T, Shaw E. Titration of trypsin, plasmin and thrombin with p-nitrophenyl-p’-guanidinobenzoate HC1. In: Methods in Enzymology 19. Perlman GE, Lorand L. (Eds) Academic Press; New York: 1970. pp 20-27
    Google Scholar
  • 20 Laskowski M. Trypsinogen and trypsin. In: Methods in Enzymology 2. Colowick SP, Kaplan NO. (Eds) Academic Press; New York: 1955. pp 26-36
    Google Scholar
  • 21 Unkeless JC, Gordon S, Reich E. Secretion of plasminogen activator by stimulated macrophages. J Exp Med 1974; 139: 834-850
    CrossrefPubMedGoogle Scholar
  • 22 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
    CrossrefPubMedGoogle Scholar
  • 23 Heussen C, Dowdle EB. Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulphate and copolymerized substrates. Anal Biochem 1980; 102: 196-202
    CrossrefPubMedGoogle Scholar
  • 24 Sumi H, Robbins KC. A functionally active heavy chain derived from human high molecular weight urokinase. J Biol Chem 1983; 258: 8014-8019
    PubMedGoogle Scholar
  • 25 Holmberg L, Bladh B, Åstedt B. Purification of urokinase by affinity chromatography. Biochim Biophys Acta 1976; 445: 215-222
    CrossrefPubMedGoogle Scholar
  • 26 Rijken DC, Collen D. Purification and characterization of the plasminogen activator secreted by human melanoma cells in culture. J Biol Chem 1981; 256: 7035-7041
    PubMedGoogle Scholar
  • 27 Burnette WN. »Western blotting«: electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 1981; 112: 195-203
    CrossrefPubMedGoogle Scholar
  • 28 Hsu S-M, Raine L, Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabelled antibody (PAP) procedures. J Histochem Cytochem 1981; 29: 577-580
    CrossrefPubMedGoogle Scholar
  • 29 Friberger P. Chromogenic peptide substrates: their use for the assay of factors in the fibrinolytic and the plasma kallikrein-kinin systems. Scand J Clin Lab Invest 1982; 42 Suppl (Suppl. 162) 9-98
    PubMedGoogle Scholar
  • 30 Chapman Jr HA, Vavrin Z, Hibbs Jr JB. Macrophage fibrinolytic activity: identification of two pathways of plasmin formation by intact cells and of a plasminogen activator inhibitor. Cell 1982; 28: 653-662
    CrossrefPubMedGoogle Scholar
  • 31 Gordon S, Cohn Z. Bacille Calmette-Guerin infection in the mouse. Regulation of macrophage plasminogen activator by T-lymphocytes and specific antigen J Exp Med 1978; 147: 1175-1187
    CrossrefPubMedGoogle Scholar
  • 32 Issernoff RR, Fusenig NE, Rifkin DB. Plasminogen activator in differentiating mouse keratinocytes. J Invest Dermatol 1983; 80: 217-222
    CrossrefPubMedGoogle Scholar
  • 33 Bykowska K, Levin EG, Rijken DC, Loskutoff DJ, Collen D. Characterization of a plasminogen activator secreted by cultured bovine aortic endothelial cells. Biochim Biophys Acta 1982; 703: 113-115
    CrossrefPubMedGoogle Scholar
  • 34 Matsuo O, Rijken DC, Collen D. Comparison of the relative fibrinogenolytic, fibrinolytic and thrombolytic properties of tissue plasminogen activator and urokinase in vitro. Throm Haemostas 1981; 45: 225-229
    PubMedGoogle Scholar
  • 35 Christman JK, Silverstein SC, Acs G. Plasminogen activators. In: Proteinases in Mammalian Cells and Tissues. Barrett AJ. (Ed) North- Holland Pub Comp; Amsterdam: 1977. pp 91-149
    Google Scholar
  • 36 Moroi M, Aoki N. Isolation and characterization of alpha2-plasmin inhibitor from human plasma. A novel proteinase inhibitor which inhibits activator-induced clot lysis J Biol Chem 1976; 251: 5956-5965
    PubMedGoogle Scholar
  • 37 Izaki S, Goldstein SM, Fukuyama K, Epstein WL. Tissue induction of a urokinase inhibitor associated with chronic granulomatous inflammation: Correlation with local fibrin deposition and clearing (in Japanese with English abstract). Blood and Vessel 1980; 11: 675-678
    PubMedGoogle Scholar
  • 38 Goldstein SM, Izaki S, Epstein WL. Inhibition of plasminogen activator associated with chronic granulomatous inflammation. Thromb Res 1979; 16: 727-735
    CrossrefPubMedGoogle Scholar
  • 39 Izaki S, Fukuyama K, Epstein WL. Modulation of antithrombin and anti-fibrinolytic activities in tissue during the development of granulomas induced by Schistosoma mansoni. J Reticuloendothel Soc 1979; 26: 507-514
    PubMedGoogle Scholar