Abstract
The major antigens secreted by the parasite Fasciola hepatica are cathepsin-L cysteine proteases. These enzymes can be isolated from the parasite excretory-secretory products in sufficient quantities for use as an antigen for the serodiagnosis of human fasciolosis. The methods illustrated in this chapter will explain the isolation of cysteine proteases from F. hepatica excretory-secretory products by gel filtration and anion exchange chromatography, and their subsequent characterization and employment in an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-fasciola antibodies in the serum of infected humans.
Key Words
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Chen, M. G. and Mott, K. E. (1990) Progress in morbidity due to Fasciola hepatica infection. Trop. Dis. Bull. 87, 1–37.
Bjorland, J., McAuley, J. B., Strauss, W., Richer, G., Antezana Eguez, A., and Hillyer, G. (1995) An outbreak of acute fasciolosis among Aymara indians in the Bolivian Altiplano. Clin. Infect. Dis. 21, 1228–1233.
Hillyer, G. V., Soler de Galanes, M., Rodriguez-Perez, J., Bjorland, J., Silva de Lagrava, M., and Ramirez Guzman, S. (1992) Use of the Falcon assay screening test-enzyme-linked immunosorbent assay (FAST-ELISA) and the enzyme-linked immnoelectro transfer blot (ETIB) to determine the prevalence of human fascioliasis in the Bolivian Altiplano. Am. J. Trop. Med. Hyg. 46, 603–609.
Stork, M. G., Venables, G. S., Jennings, S. M. F., Beesley, J. R., Bendezu, P., and Capron, A. (1973) An investigation of endemic fascioliasis in Peruvian village children. J. Trop. Med. Hyg. 76, 231–235.
Espino, A. M. and Finlay, C. M. (1994) Sandwich enzyme-linked immunosorbent assay for detection of excretory-secretory antigens in humans with fascioliasis. J. Clin. Microbiol. 32, 190–193.
Massoud, J. (1989) Fascioliasis outbreak in man and drug test (triclabendazole) in Caspian littoral, northern part of Iran. Bull. Soc. Fran. Parasitol. 8, 438–438.
Rokni, M. B., Massoud, J., O’Neill, S. M., Parkinson, M., and Dalton, J. P. (2002) Diagnosis of human fasciolosis in the Gilan province of northern Iran: application of cathepsin L-ELISA. Diagn. Microbiol. Infect. Dis. 44, 175–179.
Haseeb, A. N., el-Shazly, A. M., Arafa, M. A., and Morsy, A. T. J. (2002) A review on fascioliasis in Egypt. Egypt Soc. Parasitol. 32, 317–354.
Esteban, J. G., Gonzalez, C., Curtale, F., Munoz-Antoli, C., Valero, M. A., and Bargues, M. D. (2003) Hyperendemic fascioliasis associated with schistosomiasis in villages in the Nile Delta of Egypt. Am. J. Trop. Med. Hyg. 69, 429–437.
Tort, J., Brindley, P. J., Knox, D., Wolfe, K. H., and Dalton, J. P. (1999) Proteinases and associated genes of parasitic helminths. Adv. Parasitol. 43, 161–166.
Berasain, P., Goni, F., McGonigle, S., et al. (1997) Fasciola hepatica: parasite-secreted proteinases degrade all human IgG subclasses: determination of the specific cleavage sites and identification of the immunoglobulin fragments produced. J. Parasitol. 83, 1–5.
Smith, A., Dowd, A., McGonigle, S., et al. (1993) Purification of a cathepsin L-like proteinase secreted by adult Fasciola hepatica. Mol. Biochem. Parasitol. 62, 1–8.
Carmona, C., Dowd, A., Smith, A., and Dalton, J. P. (1993) Cathepsin L proteinase secreted by Fasciola hepatica in vitro prevents antibody-mediated eosinophil attachment to newly excysted juveniles. Mol. Biochem. Parasitol. 62, 9–18.
Brady, M. T., O’Neill, S. M., Dalton, J. P., and Mills, K. H. G. (1999) Fasciola hepatica suppresses a protective Th1 response against Bordetella pertussis. Infect. Immun. 67, 5372–5378.
O’Neill, S. M., Brady, M. T., Callanan, J. J., et al. (2000) Fasciola hepatica infection downregulates Th1 responses in mice. Parasite Immunol. 22, 147–155.
O’Neill, S. M., Mills, K. H., and Dalton, J. P. (2001) Fasciola hepatica cathepsin L cysteine proteinase suppresses Bordetella pertussis-specific interferon-gamma production in vivo. Parasite Immunol. 23, 541–547.
O’Neill, S. M., Parkinson, M., Strauss, W., Angles, R., and Dalton, J. P. (1998) Immunodiagnosis of Fasciola hepatica infection in a human population in the Bolivian Altiplano using purified cathepsin L cysteine proteinase. Am. J. Trop. Med. Hyg. 58, 417–423.
Strauss, W., O’Neill, S. M., Parkinson, M., Angles, R., and Dalton, J. P. (1999) Diagnosis of human fasciolosis: detection of anti-cathepsin L antibodies on blood samples collected on filter paper. Am. J. Trop. Med. Hyg. 60, 746–748.
Dalton, J. P. and Heffernan, M. (1989) Thiol proteinase released in vitro by Fasciola hepatica. Mol. Biochem. Parasitol. 35, 161–166.
Dowd, A. J., Smith, A. M., McGonigle, S. M., and Dalton, J. P. (1994) Purification and characterisation of a second cathepsin L proteinase secreted by the parasite trematode Fasciola hepatica. Eur. J. Biochem. 223, 91–98.
Smith, A., Dowd, A. J., Heffernan, M., Robertson, C., and Dalton, J. P. (1993) Fasciola hepatica: a secreted cathepsin L-like proteinase cleaves host immunoglobulin. Int. J. Parasitol. 23, 977–983.
Barrett, A. J. and Kirschke, H. (1980) Catheppsin B, cathepsin H and cathepsin L. Methods in Enzymol. 80, 535–561.
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of a bacteriophage T4. Nature 227, 680–685.
Acknowledgments
We would like to thank Dr. Wilma Strauss and Dr. Rene Angles (National Institute of Health, La Paz, Bolivia) for the gift of human antiserum.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
O’Neill, S.M., Mulcahy, G., Dalton, J.P. (2006). Isolation and Characterization of Cathepsin-L1 Protease From Fasciola hepatica Excretory-Secretory Products for Serodiagnosis of Human Fasciolosis. In: Adley, C.C. (eds) Food-Borne Pathogens. Methods in Biotechnology™, vol 21. Humana Press. https://doi.org/10.1385/1-59259-990-7:191
Download citation
DOI: https://doi.org/10.1385/1-59259-990-7:191
Publisher Name: Humana Press
Print ISBN: 978-1-58829-465-4
Online ISBN: 978-1-59259-990-5
eBook Packages: Springer Protocols