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Using lectins to identify hidden antigens in Fasciola hepatica

Published online by Cambridge University Press:  06 January 2011

H.C. McAllister
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, EH26 0PZ, UK
A.J. Nisbet
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, EH26 0PZ, UK
P.J. Skuce
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, EH26 0PZ, UK
D.P. Knox*
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, EH26 0PZ, UK
*

Abstract

Fasciola hepatica is the causative agent of fascioliasis, one of the most economically important helminth diseases of livestock worldwide. Traditionally, fascioliasis has been controlled by the strategic use of fasciolicidal drugs, but the emergence of resistant parasites has spurred an interest in developing vaccines as an alternative means of control. Most vaccine studies to date have evaluated conventional antigens, which are exposed to the host's immune system during the course of a natural infection. ‘Hidden’ antigens have proven to be effective vaccine candidates in other parasite species, most notably the blood-feeding nematode parasite, Haemonchus contortus, and tend to be expressed in the intestine or gut of the parasite. Fasciola hepatica is known to ingest large quantities of blood and may be vulnerable to this approach. Most, if not all, of the candidate antigens identified thus far have been membrane-bound glycoproteins which were solubilized by detergents. Here, we have attempted to employ lectins to select gut-associated glycoproteins from complex mixtures of somatic extracts of adult F. hepatica. We have conducted a comprehensive lectin-binding screen on adult histological sections with a panel of 16 fluorescently labelled lectins. Seven of the lectins bound to molecules within the gastrodermis but also bound to a range of other tissues. Within the gut tissues, jacalin and peanut lectins bound selectively to the gut lamellae and gastrodermal cells, respectively. These lectins were then used to isolate proteins from the integral membrane protein component of the adult fluke. Both lectins showed selectivity for relatively simple subsets of proteins compared to the original crude extracts.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 2011

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References

Acosta, D., Cancela, M., Piacenza, L., Roche, L., Carmona, C. & Tort, J.F. (2008) Fasciola hepatica leucine aminopeptidase, a promising candidate for vaccination against ruminant fasciolosis. Molecular and Biochemical Parasitology 158, 5264.CrossRefGoogle ScholarPubMed
Bennett, R.M. & IJpelaar, J. (2005) Updated estimates of the costs associated with 34 endemic livestock diseases in Great Britain. Journal of Agricultural Economics 56, 135144.CrossRefGoogle Scholar
Brennan, G.P., Fairweather, I., Trudgett, A., Hoey, E., McCoy, M., McConville, M., Meaney, M., Robinson, M., McFerran, N., Ryan, L., Lanusse, C., Mottier, L., Alvarez, L., Solana, H., Virkel, G. & Brophy, P.M. (2007) Understanding triclabendazole resistance. Experimental and Molecular Pathology 82, 104109.CrossRefGoogle ScholarPubMed
Brophy, P.M. & Pritchard, D.I. (1994) Parasitic helminth glutathione S-transferases: an update on their potential as targets for immuno- and chemotherapy. Experimental Parasitology 79, 8996.CrossRefGoogle ScholarPubMed
Collins, P.R., Stack, C.M., O'Neill, S.M., Doyle, S., Ryan, T., Brennan, G.P., Mousley, A., Stewart, M., Maule, A.G., Dalton, J.P. & Donnelly, S. (2004) Cathepsin L1, the major protease involved in liver fluke (Fasciola hepatica) virulence. Journal of Biological Chemistry 279, 17 03817 046.CrossRefGoogle ScholarPubMed
Creaney, J., Wijffels, G.L., Sexton, J.L., Sandeman, R.M., Spithill, T.W. & Parsons, J.C. (1995) Fasciola hepatica: localisation of glutathione S-transferase isoenzymes in adult and juvenile liver fluke. Experimental Parasitology 81, 106116.CrossRefGoogle ScholarPubMed
Dalton, J.P., Neill, S.O., Stack, C., Collins, P., Walshe, A., Sekiya, M., Doyle, S., Mulcahy, G., Hoyle, D., Khaznadji, E., Moiré, N., Brennan, G., Mousley, A., Kreshchenko, N., Maule, A.G. & Donnelly, S.M. (2003) Fasciola hepatica cathepsin L-like proteases: biology, function, and potential in the development of first generation liver fluke vaccines. International Journal of Parasitology 33, 11731181.CrossRefGoogle ScholarPubMed
Gobert, G.N., Jones, M.K. & Stenzel, D.J. (1998) Ultrastructural analysis of the adult Schistosoma japonicum by lectin cytochemistry. International Journal of Parasitology 28, 14451452.CrossRefGoogle ScholarPubMed
Hillyer, G.V. & Apt, W. (1997) Food-borne trematode infections in the Americas. Parasitology Today 61, 603609.Google Scholar
Kenyon, F., Sargison, N.D., Skuce, P.J. & Jackson, F. (2009) Sheep helminth parasitic disease in south eastern Scotland arising as a possible consequence of climate change. Veterinary Parasitology 163, 293297.CrossRefGoogle ScholarPubMed
Knox, D.P. & Smith, W.D. (2001) Vaccination against gastrointestinal nematode parasites of ruminants using gut-expressed antigens. Veterinary Parasitology 100, 2132.CrossRefGoogle ScholarPubMed
Knox, D.P., Redmond, D.L., Newlands, G.F., Skuce, P.J., Pettit, D. & Smith, W.D. (2003) The nature and prospects for gut membrane proteins as vaccine candidates for Haemonchus contortus and other ruminant trichostrongyloids. International Journal of Parasitology 33, 11291137.CrossRefGoogle ScholarPubMed
Leathem, A.J. & Brooks, S.A. (1998) Light microscopy. pp. 373in Rhodes, J.M. & Milton, J.D. (Eds) Lectin methods and protocols. New Jersey, Humana Press.Google Scholar
McGonigle, L., Mousley, A., Marks, N.J., Brennan, G.P., Dalton, J.P., Spithill, T.W., Day, T.A. & Maule, A.G. (2008) The silencing of cysteine proteases in Fasciola hepatica newly excysted juveniles using RNA interference reduces gut penetration. International Journal of Parasitology 38, 149155.CrossRefGoogle ScholarPubMed
Olsen, O.W. (1986) Fasciola hepatica. pp. 267272in Olsen, O.W. (Ed.) Animal parasites: Their life cycles and ecology. 3rd edn.New York, Dover Publications.Google Scholar
Overend, D.J. & Bowen, F.L. (1995) Resistance of Fasciola hepatica to triclabendazole. Australian Veterinary Journal 72, 275276.CrossRefGoogle ScholarPubMed
Sexton, J.L., Milner, A.R., Panaccio, M., Waddington, J., Wijfels, G., Chandler, D., Thompson, C., Wilson, L., Spithill, T.W., Mitchell, G.F. & Campbell, N.J. (1990) Glutathione S-transferase, novel vaccine against Fasciola hepatica infection in sheep. Journal of Immunology 145, 39053910.CrossRefGoogle ScholarPubMed
Smith, S.K., Pettit, D., Newlands, G.F., Redmond, D.L., Skuce, P.J., Knox, D.P. & Smith, W.D. (1999) Further immunization and biochemical studies with a protective antigen complex from the microvillar membrane of the intestine of Haemonchus contortus. Parasite Immunology 21, 187199.CrossRefGoogle ScholarPubMed
Smith, W.D., Smith, S.K. & Murray, J.M. (1994) Protection studies with integral membrane fractions of Haemonchus contortus. Parasite Immunology 16, 231241.CrossRefGoogle ScholarPubMed
Smith, W.D., Smith, S.K. & Pettit, D. (2000a) Evaluation of immunization with gut membrane glycoproteins of Ostertagia ostertagi against homologous challenge in calves and against Haemonchus contortus in sheep. Parasite Immunology 22, 239247.CrossRefGoogle ScholarPubMed
Smith, W.D., Smith, S.K., Pettit, D., Newlands, G.F. & Skuce, P.J. (2000b) Relative protective properties of three membrane glycoprotein fractions from Haemonchus contortus. Parasite Immunology 22, 6371.CrossRefGoogle ScholarPubMed
Spithill, T.W., Smooker, P.M., Sexton, J.L., Bozas, E., Morrison, C.A., Creaney, J. & Parsons, J.C. (1999) Development of vaccines against Fasciola hepatica. pp. 377401in Dalton, J.P. (Ed.) Fasciolosis. Wallingford, UK, CAB International.Google Scholar
van Dijk, J., Sargison, N.D., Kenyon, F. & Skuce, P.J. (2010) Climate change and infectious disease: helminthological challenges to farmed ruminants in temperate regions. Animal 4, 377392.CrossRefGoogle ScholarPubMed
Willadsen, P., Bird, P., Cobon, G.S. & Hungerford, J. (1995) Commercialisation of a recombinant vaccine against Boophilus microplus. Parasitology 110, S43S50.CrossRefGoogle ScholarPubMed
Wuhrer, M., Grimm, C., Dennis, R.D., Idris, M.A. & Geyer, R. (2004) The parasitic trematode Fasciola hepatica exhibits mammalian-type glycolipids as well as Gal(beta1-6)Gal-terminating glycolipids that account for cestode serological cross-reactivity. Glycobiology 14, 115126.CrossRefGoogle ScholarPubMed