Parasitology Research

, 100:131

A relevant enzyme in granulomatous reaction, active matrix metalloproteinase-9, found in bovine Echinococcus granulosus hydatid cyst wall and fluid

  • M. Marco
  • A. Baz
  • C. Fernandez
  • G. Gonzalez
  • U. Hellman
  • G. Salinas
  • A. Nieto
Original Paper

Abstract

In addition to the ability of matrix metalloproteinases (MMP) to degrade components of the extracellular matrix and their involvement in pathology-related processes of tissue remodeling, they were recently reported to enhance inflammation by activation of proinflammatory cytokines, or their release from the cell surface. In the work reported here, proteolytic activity previously found for hydatid cysts was further characterized as MMP-9. Active host MMP-9 was found in walls and fluids of bovine hydatid cysts of Echinococcus granulosus in the environment of granulomatous reaction. Pooled walls and fluids of hydatid cysts obtained from infected cattle were processed. Strong proteolytic activity was detected by zymography. The proteolytic fraction was purified by anion exchange and gelatin-agarose affinity chromatography. Major proteinases of the purified fraction were subjected to mass spectrometry and their identities were further confirmed by Western blotting using commercial anti-human MMP-9 monoclonal antibodies. Two proteinases were characterized as latent and active forms of host MMP-9. Using the same antibody for immunoblot, activity was localized, in paraffin-embedded sections of the parasite and the local host environment, to epithelioid and giant multinucleated cells. It is proposed here that MMP-9 is secreted by specialized host cells of monocytic lineage (epithelioid/giant cells) as an effector, in an attempt to digest the persistent foreign body. In vivo activation of MMP-9 suggests its involvement in inflammatory reaction and in the chemotaxis of inflammatory cells to the cyst. However, E. granulosus can deal efficiently with MMP-9. Research is suggested into possible immune evasion mechanisms, including the secretion of an inhibitory molecule.

References

  1. Bjorklund M, Koivunen E (2005) Gelatinase-mediated migration and invasion of cancer cells. Biochim Biophys Acta 1755:37–69PubMedGoogle Scholar
  2. Coltorti EA, Varela-Díaz VM (1972) IgG levels and host specificity in hydatid cyst fluid. J Parasitol 58:753PubMedCrossRefGoogle Scholar
  3. Coltorti EA, Varela-Díaz VM (1974) Echinococcus granulosus: penetration of macromolecules and their localization on the parasite membranes of cysts. Exp Parasitol 35:225–231CrossRefGoogle Scholar
  4. Chordi A, Kagan IG (1965) Identification and characterization of antigenic components of sheep hydatid fluid by immunoelectrophoresis. J Parasitol 51:63PubMedCrossRefGoogle Scholar
  5. Craig PS (1988) Immunology of human hydatid disease. In: Grimwade AM (ed) ISI atlas of science: immunology. Institute for Scientific Information, Philadelphia, p 95Google Scholar
  6. Díaz A, Ferreira AM, Sim RB (1997) Complement evasion by Echinococcusgranulosus: sequestration of host factor H in the hydatid cyst wall. J Immunol 158:3779–3786PubMedGoogle Scholar
  7. Díaz A, Ibarguren S, Breijo M, Willis A, Sim R (2000a) Host-derived annexin II at the host–parasite interface of the Echinococcus granulosus hydatid cyst. Mol Biochem Parasitol 110:171–176PubMedCrossRefGoogle Scholar
  8. Díaz A, Willis A, Sim R (2000b) Expression of the proteinase specialized in bone resorption, cathepsin K, in granulomatous inflammation. Mol Med 6(8):648–659PubMedGoogle Scholar
  9. Eckert J, Deplazes P (2004) Biological, epidemiological, and clinical aspects of Echinococcosis a zoonosis of increasing concern. Clin Microbiol Rev 17(1):107–135 (Jan)PubMedCrossRefGoogle Scholar
  10. Egeblad M, Werb Z (2002) New functions for the matrix metalloproteinases in cancer progression. Nat Rev 2:161–174Google Scholar
  11. Fernández C (1992) Studies on the glutathione S-transferase(s) from Echinococcus granulosus protoscoleces. Doctoral dissertation, University of Cambridge, Cambridge, UKGoogle Scholar
  12. Fridman R, Toth M, Pena D, Mobashery S (1995) Activation of progelatinase B (MMP-9) by gelatinase A (MMP-2). Cancer Res 55:2548–2555PubMedGoogle Scholar
  13. Fridman R, Toth M, Chvyrkova I, Meroush O, Mobashery S (2003) Cell surface association of matrix metalloproteinase-9 (gelatinase B). Cancer Metastasis Rev 22:153–166PubMedCrossRefGoogle Scholar
  14. Gómez D, De Lorenzo M, Alonso D, Andrade Z (1999) Expression of metalloproteinases (MMP-1, MMP-2 and MMP-9) and their inhibitors (TIMP-1 and TIMP-2) in schistosomal portal fibrosis. Am J Trop Med Hyg 61(1):9–13 (Jul)PubMedGoogle Scholar
  15. Heath DD (1995) Immunology and Echinococcus infections. In: Thompson RCA, Lymberly AJ (eds) Echinococcus and hydatid disease. CAB International, Wallingford, UK, p 183Google Scholar
  16. Izzo A, Izzo L, Kasimos J, Majka S (2004) A matrix metalloproteinase inhibitor promotes granuloma formation during early phase of Mycobacterium tuberculosis pulmonary infection. Tuberculosis 84:387–396PubMedCrossRefGoogle Scholar
  17. Kagan IG, Norman L (1963) The isolation and characterization of two host antigens in hydatid cyst fluid of Echinococcus granulosus. Am J Trop Med Hyg 12:346PubMedGoogle Scholar
  18. Kovaleva ES, Masler EP, Skantar AM, Chitwood DJ (2004) Novel matrix metalloproteinase from the cyst nematodes Heterodera glycines and Globodera rostochiensis. Mol Biochem Parasitol 136(1):109–112 (Jul)PubMedCrossRefGoogle Scholar
  19. Lightowlers MW (1990) Immunology and molecular biology of Echinococcus infections. Int J Parasitol 20:471PubMedCrossRefGoogle Scholar
  20. Lightowlers MW, Gottstein B (1995) Echinococcosis/hydatidosis: antigens, immunological and molecular diagnosis. In: Thompson RCA, Lymbery AJ (eds) Echinococcus and hydatid disease. CAB International, Wallingford UK, pp 355–410Google Scholar
  21. Marco M, Nieto A (1991) Metalloproteinases in the larvae of Echinococcus granulosus. Int J Parasitol 21(6):743–746PubMedCrossRefGoogle Scholar
  22. Matthews BE (1982) Behaviour and enzyme release by Anisakis sp larvae (Nematoda: Ascaridida). J Helminthol 56:177–183PubMedCrossRefGoogle Scholar
  23. McCawley LJ, Matrisian LM (2001) Matrix metalloproteinases: they’re not just for matrix anymore! Curr Opin Cell Biol 13:534–540PubMedCrossRefGoogle Scholar
  24. McKerrow (1989) Parasite proteases. Exp Parasitol 68:111–115PubMedCrossRefGoogle Scholar
  25. McKerrow JH, Doenhoff MJ (1988) Schistosoma proteases. Parasitol Today 12:334–449CrossRefGoogle Scholar
  26. McKerrow JH, Brindley P, Brown M, Gam AA, Stunton C, Neva FA (1990) Strongyloides stercoralis: identification of a protease that facilitates penetration of skin by the infective larvae. Exp Parasitol 70:134–143PubMedCrossRefGoogle Scholar
  27. Morris SR, Sakanari JA (1994) Characterization of the serine protease and serine protease inhibitor from the tissue-penetrating nematode Anisakis simplex. J Biol Chem 269:27650–27656PubMedGoogle Scholar
  28. Nagase H (1997) Activation mechanisms of matrix metalloproteinases. Biol Chem 378(3–4):151–160 (Mar)PubMedGoogle Scholar
  29. Nagase H, Woesner JF (1999) Matrix metalloproteinases. J Biol Chem 274:21491–21494PubMedCrossRefGoogle Scholar
  30. Okada Y, Gonoji Y, Naka K, Tomita K, Nakanishi I, Iwata K, Yamashita K, Hayakawa T (1992) Matrix metalloproteinase 9 (92-kDa gelatinase/type IV collagenase) from HT 1080 human fibrosarcoma cells. Purification and activation of the precursor and enzymic properties. J Biol Chem 267(30):21712–21719 (Oct 25)PubMedGoogle Scholar
  31. Opdenakker G, Van den Steen P, Van Damme J (2001) Gelatinase B: a tuner and amplifier of immune functions. Trends Immunol 22(10):571–579 (Oct)PubMedCrossRefGoogle Scholar
  32. Opdenakker G, Nelissen I, Van Damme J (2003) Functional roles and therapeutic targeting of gelatinase B and chemokines in multiple sclerosis. Lancet Neurol 2:747–756 (Dec)PubMedCrossRefGoogle Scholar
  33. Parks W, Wilson C, Lopez-Boado Y (2004) Matrix metalloproteinases as modulators of inflammation and innate immunity. Nat Rev Immunol 4:617–629 (Aug)PubMedCrossRefGoogle Scholar
  34. Petralanda I, Yarzabal L, Piessens WF (1986) Studies on a filarial antigen with collagenase activity. Mol Biochem Parasitol 19:51–59PubMedCrossRefGoogle Scholar
  35. Quiding-Jarbrink M, Smith D, Bancroft G (2001) Production of matrix metalloproteinases in response to mycobacterial infection. Infect Immun 69(9):5661–5670 (Sep)PubMedCrossRefGoogle Scholar
  36. Rasheed S, Nelson-Rees W, Toth E, Arnstein P, Gardner M (1974) Characterization of a newly derived human sarcoma cell line (HT1080). Cancer 33:1027–1033PubMedCrossRefGoogle Scholar
  37. Robertson BD, Bianco AT, Mc Kerrow JH, Maizels RM (1989) Toxocara canis: proteolytic enzymes secreted by the infective larvae in vitro. Exp Parasitol 69:30–36PubMedCrossRefGoogle Scholar
  38. Shapiro SZ, Bahr GM, Hira PR (1992) Analysis of host components in hydatid cyst fluid and immunoblot diagnosis of human Echinococcus granulosus infection. Ann Trop Med Parasitol 86:503PubMedGoogle Scholar
  39. Taggart C, Greene C, Carrol T, O’Neill S, McElvaney N (2005) Elastolytic proteases, inflammation resolution and deregulation in chronic infective lung disease. Am J Respir Crit Care Med 171:1070–1076PubMedCrossRefGoogle Scholar
  40. Thomas J, Kothare SN (1975) Tissue response in hydatidosis. Indian J Med Res 63(12):1761–1766 (Dec)PubMedGoogle Scholar
  41. Thompson RCA (1995) Biology and systematics of Echinococcus. In: Thompson RCA, Lymbery AJ (eds) Echinococcus and hydatid disease. CAB International, Wallingford, UK, pp 1–50Google Scholar
  42. Towbin H, Stahelin T, Gordon J (1979) Electroforetic transfer of protein from polyacrylamide gels to nitrocellulose sheets. Proc Natl Acad Sci USA 76:4350PubMedCrossRefGoogle Scholar
  43. Trancart MM, Chalmeigne N, Girardot C, Zarpanelian C, Prigent D (1992) Gelatinase is the main matrix metalloproteinase involved in granuloma-induced cartilage degradation. Int J Tissue React 14(6):287–294PubMedGoogle Scholar
  44. Turk J (1989) Current status review: a comparison of secretory epithelioid cells and phagocytosing macrophages in experimental mycobacterial granulomas. Br J Path 70:589–596Google Scholar
  45. Uparanukraw P, Morakote N, Harnnoi T, Dantrakool A (2001) Molecular cloning of a gene encoding matrix metalloproteinase-like protein from Gnathostoma spinigerum. Parasitol Res 87:751–757PubMedCrossRefGoogle Scholar
  46. Van den Steen P, Dubois B, Nelissen I, Rudd P, Dwek R, Opdenakker G (2002) Biochemistry and molecular biology of gelatinase B or matrix metalloproteinase-9 (MMP-9). Crit Rev Biochem Mol Biol 37(6):375–536PubMedCrossRefGoogle Scholar
  47. Varela-Díaz VM, Coltorti EA (1972) Further evidence of the passage of host immunoglobulins into hydatid cysts. J Parasitol 58:1015–1016PubMedCrossRefGoogle Scholar
  48. Varela-Díaz VM, Coltorti EA (1973) The presence of host immunoglobulins in hydatid cyst membranes. J Parasitol 59:484–488PubMedCrossRefGoogle Scholar
  49. Wada K, Sato H, Kinoh H, Kajita M, Yamamoto H, Seiki M (1998) Cloning of three Caenorhabditis elegans genes potentially encoding novel matrix metalloproteinases. Gene 211:57–62PubMedCrossRefGoogle Scholar
  50. Williams G, Williams W (1983) Granulomatous inflammation—a review. J Clin Pathol 36:723–733PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • M. Marco
    • 1
  • A. Baz
    • 1
  • C. Fernandez
    • 1
  • G. Gonzalez
    • 1
  • U. Hellman
    • 2
  • G. Salinas
    • 1
  • A. Nieto
    • 1
  1. 1.Cátedra de Inmunología, Facultad de Química, Facultad de Ciencias, Instituto de HigieneUniversidad de la RepúblicaMontevideoUruguay
  2. 2.Ludwig Institute for Cancer Research (Uppsala Branch)UppsalaSweden

Personalised recommendations