Inflammation Research

, Volume 44, Issue 12, pp 529–534 | Cite as

Prevention of acute autoimmune encephalomyelitis and abrogation of relapses in murine models of multiple sclerosis by the protease inhibitor D-penicillamine

  • K. Norga
  • L. Paemen
  • S. Masure
  • C. Dillen
  • H. Heremans
  • A. Billiau
  • H. Carton
  • L. Cuzner
  • T. Olsson
  • J. Van Damme
  • G. Opdenakker


Thein vitro activity of gelatinase B, an enzyme whose appearance in the cerebrospinal fluid is associated with inflammatory diseases of the central nervous system, was dose-dependently inhibited by the antirheumatic D-penicillamine. Inhibition of gelatinase B in electrophoretically pure preparations and in cell culture supernatants and human body fluids was obtained at dosages reached in the circulation of patients treated with a peroral dosis of 750mg D-penicillamine per day. In mice, developing acute demyelination, D-penicillamine significantly reduced the mortality and morbidity rates of experimental allergic encephalomyelitis (EAE). In chronic relapsing EAE in Biozzi AB/H mice, an animal model for relapses in multiple sclerosis (MS), it attenuated the exacerbations, even when the treatment was started after the primary full-blown disease had developed. We infer protease inhibition as the mechanism of action of D-penicillamine and suggest that its use may be effective as peroral treatment for MS.

Key words

Matrix metalloproteinases D-penicillamine Experimental allergic encephalomyelitis Multiple sclerosis Demyelination 



cerebrospinal fluid


experimental allergic encephalomyelitis




matrix metalloproteinase


demultiple sclerosis


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. [1]
    Woessner JF. Matrix metalloproteinases and their inhibitors in connective tissue remodeling. FASEB J 1991;5:2145–54.PubMedGoogle Scholar
  2. [2]
    Opdenakker G, Masure S, Grillet B, Van Damme J. Cytokinemediated regulated of human leukocyte gelatinases and role in arthritis. Lymphokine Cytokine Res 1991;10:317–24.PubMedGoogle Scholar
  3. [3]
    Gijbels K, Masure S, Carton H, Opdenakker G. Gelatinase in the cerebrospinal fluid of patients with multiple sclerosis and other inflammatory neurological diseases. J Neuroimmunol 1992;41:29–34.PubMedGoogle Scholar
  4. [4]
    Paemen L, Olsson T, Söderström M, Van Damme J, Opdenakker G. Evaluation of gelatinases and IL-6 in the cerebrospinal fluid of patients with optic neuritis, multiple sclerosis and other inflammatory neurological diseases. Eur J Neurol 1994;1:55–63.Google Scholar
  5. [5]
    Proost P, Van Damme J, Opdenakker G. Leukocyte gelatinase B cleavage releases encephalitogens from human myelin basic protein. Biochem Biophys Res Commun 1993;192:1175–81.PubMedGoogle Scholar
  6. [6]
    Gijbels K, Proost P, Masure S, Carton H, Billiau A, Opdenakker G. Gelatinase B is present in the cerebrospinal fluid during experimental autoimmune encephalomyelitis and cleaves myelin basic protein. J Neurosci Res 1993;36:432–40.PubMedGoogle Scholar
  7. [7]
    Masure S, Proost P, Van Damme J, Opdenakker G. Purification of 91-kDa neutrophil gelatinase. Release by the activating peptide interleukin-8. Eur J Biochem 1991;198:391–8.PubMedGoogle Scholar
  8. [8]
    Proost P, De Wolf-Peeters C, Conings R, Opdenakker G, Billiau A, Van Damme J. Identification of a novel granulocyte chemotactic protein (GCP-2) from human tumor cells. J Immunol 1992;150:1000–10.Google Scholar
  9. [9]
    Montgomery AMP, Sabzevari H, Reisfeld RA. Production and regulation of gelatinase B by human T-cells. Biochim Biophys Acta 1993;1176:265–8.PubMedGoogle Scholar
  10. [10]
    Edwards DR, Murphy G, Reynolds JJ, Whitham SE, Docherty AJP, Angel P, et al. Transforming growth factor beta modulates the expression of collagenase and metalloproteinase inhibitor. EMBO J 1987;6:1899–904.PubMedGoogle Scholar
  11. [11]
    Van Damme J, Proost P, Lenaerts J-P, Opdenakker G. Structural and functional identification of two human, tumor-derived monocyte chemotactic proteins (MCP-2 and MCP-3) belonging to the chemokine family. J Exp Med 1992;176:59–65.PubMedGoogle Scholar
  12. [12]
    Opdenakker G, Van Damme J. Cytokine-regulated proteases in autoimmune diseases. Immunol Today 1994;15:103–7.PubMedGoogle Scholar
  13. [13]
    Roche PC, Campeau JD, Shaw ST. Comparative electrophoretic analysis of human and porcine plasminogen activators in SDS-polyacrylamide gels containing plasminogen and casein. Biochim Biophys Acta 1983;745:82–9.PubMedGoogle Scholar
  14. [14]
    Wilhelm SM, Collier IE, Marmer BL, Eisen AZ, Grant GA, Goldberg GI. SV40-transformed human lung fibroblasts secrete a 92-kDa type IV collagenase which is identical to that secreted by normal human macrophages. J Biol Chem 1989;264:17213–21.PubMedGoogle Scholar
  15. [15]
    Houde M, De Bruyne G, Bracke M, Ingelman-Sundberg M, Skoglund G, Masure S, et al. Differential regulation of gelatinase B and tissue-type plasminogen activator expression in human Bowes melanoma cells. Int J Cancer 1993;53:395–400.PubMedGoogle Scholar
  16. [16]
    Baker D, O'Neill JK, Davison AN, Turk JL. Control of immune-mediated disease of the central nervous system requires the use of a neuroactive agent: elucidation by the action of mitoxantrone. Clin Exp Immunol 1992;90:124–8.PubMedGoogle Scholar
  17. [17]
    Jaffe IJ. Penicillamine. In: Kelly, Harris, Ruddy, Sledge, editors. Textbook of Rheumatology. 4th ed. W. B. Saunders Company, 1994;763–4.Google Scholar
  18. [18]
    Tremblay P, Houde M, Arbour N, Rochefort D, Masure S, Mandeville R, et al. Differential effects of PKC inhibitors on gelatinase B and interleukin-6 production in the mouse macrophage. Cytokine 1995;7:130–6.PubMedGoogle Scholar
  19. [19]
    Opdenakker G, Van Damme J. Cytokines and proteases in invasive processes: molecular similarities between inflammation and cancer. Cytokine 1992;4:251–8.PubMedGoogle Scholar
  20. [20]
    Rinne UK, Riekkinen P. Esterase, peptidase and proteinase activities of human cerebrospinal fluid in multiple sclerosis. Acta Neurol Scand 1968;44:156–67.PubMedGoogle Scholar
  21. [21]
    Cuzner ML, Davison AN, Rudge P. Proteolytic enzyme activity of blood leukocytes and cerebrospinal fluid in multiple sclerosis. Ann Neurol 1978;4:337–44.PubMedGoogle Scholar
  22. [22]
    Brosnan CF, Cammer W, Norton WT, Bloom BR. Proteinase inhibitors suppress the development of experimental allergic encephalomyelitis. Nature 1980;285:235–7.PubMedGoogle Scholar
  23. [23]
    Chantry A, Gregson N, Glynn P. Degradation of myelin basic protein by a membrane-associated metalloprotease: neural distribution of the enzyme. Neurochem Res 1990;17:861–7.Google Scholar
  24. [24]
    McGeehan GM, Becherer JD, Bast Jr RC, Boyer CM, Champion B, Connolly KM, et al. Regulation of tumour necrosis factor-alfa processing by a metalloproteinase inhibitor. Nature 1994;370:558–61.PubMedGoogle Scholar
  25. [25]
    Gearing AR, Beckett P, Christodoulou M, Churchill M, Clements J, Davidson AH, et al. Processing of tumour necrosis factor-alfa precursor by metalloproteinases. Nature 1994;370:555–7.PubMedGoogle Scholar
  26. [26]
    Mohler KM, Sleath PR, Fitzner JN, Cerretti DP, Alderson M, Kerwar SS, et al. Protection against a lethal dose of endotoxin by an inhibitor of tumour necrosis factor processing. Nature 1994;370:218–20.PubMedGoogle Scholar
  27. [27]
    Lefebvre V, Peeters-Joris C, Vaes G. Production of gelatindegrading matrix metalloproteinases (type IV collagenases) and inhibitors by articular chondrocytes during their dedifferentiation by serial subcultures and under stimulation by interleukin-1 and tumor necrosis factor-alpha. Biochim Biophys Acta 1991;1094:8–18.PubMedGoogle Scholar
  28. [28]
    Partridge CA, Jeffrey JJ, Malik AB. A 96kDa gelatinase induced by TNF-alpha contributes to increased microvascular endothelial permeability. Am J Physiol 1993;265:L438–47.PubMedGoogle Scholar
  29. [29]
    Raine CS. Multiple sclerosis: TNF revisited, with promise. Nature Med 1995;1:211–3.PubMedGoogle Scholar
  30. [30]
    Zamvil SS, Steinman L. The T lymphocyte in experimental allergic encephalomyelitis. Ann Rev Immunol 1990;8:579–621.Google Scholar
  31. [31]
    Cohen IR, Mor F. On the regulation of EAE. Int Rev Immunol 1993;9:243–9.PubMedGoogle Scholar
  32. [32]
    Fabry Z, Raine CS, Hart MN. Nervous tissue as an immune compartment: the dialect of the immune response in the CNS. Immunol Today 1994;15:218–24.PubMedGoogle Scholar
  33. [33]
    Zhang J, Medaer R, Stinissen P, Hafler D, Raus J. MHC-restricted depletion of human myelin basic protein-reactive T cells by T cell vaccination. Science 1993;261:1451–4.PubMedGoogle Scholar
  34. [34]
    Waldor MK, Sriram S, Hardy R, Herzenberg LA, Lanier L, Lim M, et al. Reversal of experimental allergic encephalomyelitis with monoclonal antibody to a T-cell subset marker. Science 1985;227:415–7.PubMedGoogle Scholar
  35. [35]
    Lindsey JW, Hodgkinson S, Mehta R, Siegel RC, Mitchell DJ, Lim M, et al. Phase 1 clinical trial of chimeric monoclonal anti-CD4 antibody in multiple sclerosis. Neurol 1994;44:413–9.Google Scholar
  36. [36]
    Whitacre CC, Gienapp IE, Orosz CG, Bitar D. Oral tolerance in experimental autoimmune encephalomyelitis. III. Evidence for clonal anergy. J Immunol 1991;147:2155–63.PubMedGoogle Scholar
  37. [37]
    Weiner HL, Mackin GA, Matsui M, Orav EJ, Khoury SJ, Dawson DM, et al. Double-blind pilot trial of oral tolerization with myelin antigens in multiple sclerosis. Science 1993;259:1321–4.PubMedGoogle Scholar
  38. [38]
    Adorini L, Muller S, Cardinaux F, Lehmann PV, Falcioni F, Nagy ZA. In vivo competition between self peptides and foreign antigens in T-cell activation. Nature 1988;334:623–35.PubMedGoogle Scholar
  39. [39]
    Jameson BM, McDonnell JM, Marini JC, Korngold R. A rationally designed CD4 analogue inhibits experimental allergic encephalomyelitis. Nature 1994;368:744–6.PubMedGoogle Scholar
  40. [40]
    Sommer N, Loschmann PA, Northoff GH, Weller M, Steinbrecher A, Steinbach JP, et al. The antidepressant rolipram suppresses cytokine production and prevents autoimmune encephalomyelitis. Nature Med 1995;1:244–8.PubMedGoogle Scholar
  41. [41]
    Gijbels K, Galardy RE, Steinman L. Reversal of experimental autoimmune encephalomyelitis with a hydroxamate inhibitor of matrix metalloproteases. J Clin Invest 1994;94:2177–82.PubMedGoogle Scholar
  42. [42]
    Hewson AK, Smith TS, Leonard JP, Cuzner ML. Suppression of experimental allergic encephalomyelitis by the matrix metalloproteinase inhibitor Ro31-9790. Inflamm Res 1995;44:345–9.PubMedGoogle Scholar

Copyright information

© Birkhäuser Verlag, Basel 1995

Authors and Affiliations

  • K. Norga
    • 1
  • L. Paemen
    • 1
  • S. Masure
    • 1
  • C. Dillen
    • 2
  • H. Heremans
    • 2
  • A. Billiau
    • 2
  • H. Carton
    • 3
  • L. Cuzner
    • 4
  • T. Olsson
    • 5
  • J. Van Damme
    • 1
  • G. Opdenakker
    • 1
  1. 1.Laboratory of Molecular ImmunologyUniversity of LeuvenLeuvenBelgium
  2. 2.Laboratory of Immunobiology, Rega Institute for Medical ResearchUniversity of LeuvenLeuvenBelgium
  3. 3.Department of NeurologyLeuven University Medical SchoolLeuvenBelgium
  4. 4.Institute of Neurology, Multiple Sclerosis LaboratoryUniversity of LondonLondonUK
  5. 5.Department of NeurologyKarolinska Institute and Huddinge University HospitalHuddingeSweden

Personalised recommendations