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Characterization of proteinases in different isolates of adult Haemonchus contortus

Published online by Cambridge University Press:  01 November 2004

D. L. REDMOND  and
R. WINDHAM
D. L. REDMOND
Affiliation:
Moredun Research Institute, International Research Centre, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 0PZ, Scotland, UK
R. WINDHAM
Affiliation:
Moredun Research Institute, International Research Centre, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 0PZ, Scotland, UK
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Abstract

A high degree of intra- and inter-geographical variation has been demonstrated previously in the excretory/secretory proteinases released by adult Haemonchus contortus. Proteinase activity has also been associated with host-protective ‘hidden’ antigens isolated from the gut of adult H. contortus. If similar geographical strain variation also exists within the gut-associated proteinases, this will have important implications for the development of a globally effective vaccine. The proteinases active in integral-membrane protein extracts from 3 different strains of adult H. contortus were characterized on the basis of their pH optima and molecular size. Although enzyme activity was detected over a wide pH range, the majority of proteinase activity was detected at acidic pH. Differences in specific activity and size of enzymes were observed between the 3 different parasite strains at different pH values. A high degree of conservation in reactive peptides was observed when protein extracts were probed with antisera raised to the protective hidden gut-antigen complexes isolated from the Moredun strain of H. contortus, or to bacterially expressed subcomponents thereof. Therefore, despite the observed differences in membrane-bound proteinase profiles, the similarity of the immunogenic response against these hidden antigens may be sufficient to prove protective against different geographical isolates of H. contortus.

Keywords

Haemonchus contortusproteinasesvaccinationstrain divergence
Type
Research Article
Information
Parasitology , Volume 130 , Issue 4 , April 2005 , pp. 429 - 435
Copyright
2005 Cambridge University Press

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References

REFERENCES

JACKSON, F. & COOP, R. L. ( 2000). The development of anthelmintic resistance in sheep nematodes. Parasitology 120 (Suppl.) S95–S107.CrossRefGoogle Scholar
KABAGAMBE, E. K., BARRAS, S. R., LI, Y., PENA, M. T., SMITH, W. D. & MILLER, J. E. ( 2000). Attempts to control haemonchosis in grazing ewes by vaccination with gut membrane proteins of the parasite. Veterinary Parasitology 92, 15–23.CrossRefGoogle Scholar
KARANU, F. N., RURANGIRWA, F. R., McGUIRE, T. C. & JASMER, D. P. ( 1993). Haemonchus contortus: identification of proteases with diverse characteristics in adult worm excretory-secretory products. Experimental Parasitology 77, 362–371.CrossRefGoogle Scholar
KARANU, F. N., RURANGIRWA, F. R., McGUIRE, T. C. & JASMER, D. P. ( 1997). Haemonchus contortus: Inter- and intrageographic isolate heterogeneity of proteases in adult worm excretory-secretory products. Experimental Parasitology 86, 89–91.CrossRefGoogle Scholar
KNOX, D. P. & KENNEDY, M. W. ( 1988). Proteinases released by the parasitic larval stages of Ascaris suum, and their inhibition by antibody. Molecular and Biochemical Parasitology 28, 207–216.CrossRefGoogle Scholar
KNOX, D. P., REDMOND, D. L. & JONES, D. G. ( 1993). Characterization of proteinases in extracts of adult Haemonchus contortus, the ovine abomasal parasite. Parasitology 106, 395–404.CrossRefGoogle Scholar
KNOX, D. P., SMITH, S. K. & SMITH, W. D. ( 1999). Immunization with an affinity purified extract from the adult parasite protects lambs against infection with Haemonchus contortus. Parasite Immunology 21, 201–210.CrossRefGoogle Scholar
LONGBOTTOM, D., REDMOND, D. L., RUSSELL, M., LIDDELL, S., SMITH, W. D. & KNOX, D. P. ( 1997). Molecular cloning and characterisation of a putative aspartate proteinase associated with a gut membrane protein complex from adult Haemonchus contortus. Molecular and Biochemical Parasitology 88, 63–72.CrossRefGoogle Scholar
MUNN, E. A. ( 1997). Rational design of nematode vaccines: hidden antigens. International Journal for Parasitology 27, 359–366.CrossRefGoogle Scholar
MUNN, E. A., SMITH, T. S., SMITH, H., JAMES, F. M., SMITH, F. C. & ANDREWS, S. J. ( 1997). Vaccination against Haemonchus contortus with denatured forms of the protective antigen H11. Parasite Immunology 19, 243–248.CrossRefGoogle Scholar
OTSEN, M., PLAS, M. E., LENSTRA, J. A., ROOS, M. H. & HOEKSTRA, R. ( 2000). Microsatellite diversity of isolates of the parasitic nematode Haemonchus contortus. Molecular and Biochemical Parasitology 110, 69–77.CrossRefGoogle Scholar
REDMOND, D. L. & KNOX, D. P. ( 2004). Protection studies in sheep using affinity-purified and recombinant cysteine proteinases of adult Haemonchus contortus. Vaccine 22, 4252–4261.CrossRefGoogle Scholar
REDMOND, D. L., KNOX, D. P., NEWLANDS, G. & SMITH, W. D. ( 1997). Molecular cloning and characterisation of a developmentally regulated putative metallopeptidase present in a host protective extract of Haemonchus contortus. Molecular and Biochemical Parasitology 85, 77–87.CrossRefGoogle Scholar
REHMAN, A. & JASMER, D. P. ( 1998). A tissue specific approach for analysis of membrane and secreted protein antigens from Haemonchus contortus gut and its application to diverse nematode species. Molecular and Biochemical Parasitology 97, 55–68.CrossRefGoogle Scholar
ROOS, M. H., OSTEN, M., HOEKSTRA, R., VEENSTRA, J. G. & LENSTRA, J. A. ( 2004). Genetic analysis of inbreeding of two strains of the parasitic nematode Haemonchus contortus. International Journal for Parasitology 34, 109–115.CrossRefGoogle Scholar
RUIZ, A., MOLINA, J. M., NJUE, A. & PRICHARD, R. K. ( 2004). Genetic variability in cysteine proteinase genes of Haemonchus contortus. Parasitology 128, 549–559.CrossRefGoogle Scholar
SKUCE, P. J., REDMOND, D. L., LIDDELL, S., STEWART, E. M., NEWLANDS, G. F. J., SMITH, W. D. & KNOX, D. P. ( 1999). Molecular cloning and characterization of gut-derived cysteine proteinases associated with a host protective extract from Haemonchus contortus. Parasitology 119, 405–412.CrossRefGoogle Scholar
SMITH, S. K. & SMITH, W. D. ( 1996). Immunisation of sheep with an integral membrane glycoprotein complex of Haemonchus contortus and with its major polypeptide components. Research in Veterinary Science 60, 1–6.CrossRefGoogle Scholar
SMITH, S. K., PETTIT, D., NEWLANDS, G. F. J., REDMOND, D. L., SKUCE, P. J., KNOX, D. P. & SMITH, W. D. ( 1999). Further immunisation and biochemical studies with a protective antigen complex from the microvillar membrane of the intestine of Haemonchus contortus. Parasite Immunology 21, 187–199.CrossRefGoogle Scholar
SMITH, T. S., GRAHAM, M., MUNN, E. A., NEWTON, S. E., KNOX, D. P., COADWELL, W. J., McMICHAEL-PHILIPS, D., SMITH, H. & OLIVER, J. J. ( 1997). Cloning and characterization of a microsomal aminopeptidase from the intestine of the nematode Haemonchus contortus. Biochimica et Biophysica Acta 1338, 295–306.CrossRefGoogle Scholar
SMITH, T. S., MUNN, E. A., GRAHAM, M., TAVERNOR, A. S. & GREENWOOD, C. A. ( 1993). Purification and evaluation of the integral membrane protein H11 as a protective antigen against Haemonchus contortus. International Journal for Parasitology 23, 271–280.CrossRefGoogle Scholar
SMITH, W. D. & SMITH, S. K. ( 1993). Evaluation of aspects of the protection afforded to sheep immunised with a gut membrane protein of Haemonchus contortus. Research in Veterinary Science 55, 1–9.CrossRefGoogle Scholar
SMITH, W. D., SMITH, S. K. & MURRAY, J. M. ( 1994). Protection studies with integral membrane fractions of Haemonchus contortus. Parasite Immunology 16, 231–241.CrossRefGoogle Scholar
SMITH, W. D., VAN WYK, J. A. & VAN STRIJP, M. F. ( 2001). Preliminary observations on the potential of gut membrane proteins of Haemonchus contortus as candidate vaccine antigens in sheep on naturally infected pasture. Veterinary Parasitology 98, 285–297.CrossRefGoogle Scholar
SYKES, A. R. ( 1994). Parasitism and production in farm animals. Animal Production 59, 155–172.CrossRefGoogle Scholar
VAN WYK, J. A., MALAN, F. S., GERDER, H. M. & ALVES, R. M. ( 1987). Two field strains of Haemonchus contortus resistant to rafoxanide. Journal of Veterinary Research 54, 143–146.Google Scholar
WILLIAMSON, A. L., BRINDLEY, P. J., KNOX, D. P., HOTEZ, P. J. & LOUKAS, A. ( 2003). Digestive proteases of blood-feeding nematodes. Trends in Parasitology 19, 417–423.CrossRefGoogle Scholar
WOODWARD, M. P., YOUNG, W. W. Jr. & BLOODGOOD, R. A. ( 1985). Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation. Journal of Immunological Methods 78, 143–153.CrossRefGoogle Scholar