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Lectin binding to secretory structures, the cuticle and the surface coat of Toxocara canis infective larvae

Published online by Cambridge University Press:  06 April 2009

A. P. Page ,
W. Rudin  and
R. M. Maizels
A. P. Page
Affiliation:
Wellcome Research Centre for Parasitic Infections, Department of Biology, Imperial College of Science, Technology and Medicine, Prince Consort Road, London SW7 2BB
W. Rudin
Affiliation:
Wellcome Research Centre for Parasitic Infections, Department of Biology, Imperial College of Science, Technology and Medicine, Prince Consort Road, London SW7 2BB
R. M. Maizels
Affiliation:
Wellcome Research Centre for Parasitic Infections, Department of Biology, Imperial College of Science, Technology and Medicine, Prince Consort Road, London SW7 2BB
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Summary

Toxocara canis infective larvae are known to produce abundant glycosylated molecules which may be found associated with the surface or secreted into their environment. Using a range of fluorescein-conjugated and gold-conjugated lectins, the localization of particular carbohydrates was defined on the surface of live parasites, and internally at the ultrastructural level. Surface exposure of N-acetyl galactosamine and N-acetyl glucosamine was deduced by binding of FITC-conjugated Helix pomatia (HPA) and wheat-germ agglutinins (WGA). These sugars appear to be associated with a densely staining surface coat as conventional immuno-electron microscopy procedures dissipate this coat and reveal no surface binding site for these lectins. However, by using cryo-immuno-electron microscopical (C-IEM) techniques, the surface coat is retained and can be shown to bind WGA. The fluorescent lectins also revealed strong WGA binding to the secretory and amphidial pores, while the buccal opening and the cuticular alae bound HPA. Corresponding results were obtained at the ultrastructural level. Thus, HPA bound to the electron-dense area of the cuticle, areas of local cuticular thickening such as the alae and buccal labia, as well as to the oesophageal lumen. WGA also bound to the thickened cuticle of the alae and the buccal opening, but showed no reaction to either the electron-dense layer of the cuticle or the oesophageal lumen. Unlike HPA, WGA did bind specifically to the secretory column contents and the electron-dense regions of the lips associated with the chemosensory amphids. The compartmentalization of the sugars N-acetyl galactosamine and N-acetyl glucosamine, their sources and routes of surface expression and the possible association with the TES glycoprotein antigens are discussed.

Keywords

lectinssecretory organssurface coatcuticle
Type
Research Article
Information
Parasitology , Volume 105 , Issue 2 , October 1992 , pp. 285 - 296
Copyright
Copyright © Cambridge University Press 1992

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References

REFERENCES

Badley, J. E., Grieve, R. B., Bowman, D. D., Glickman, L. T. & Rockey, J. H. (1987). Analysis of Toxocara canis larval excretory–secretory antigens: physicochemical characterization and antibody recognition. Journal of Parasitology 73, 593–600.CrossRefGoogle ScholarPubMed
Beaver, P. C. (1966). Zoonoses, with particular reference to parasites of veterinary importance. In Biology of Parasites, pp. 215–227. New York: Academic Press.Google Scholar
Betschart, B. (1990). Structure, molecular biology and immunology of the cuticle of parasitic nematodes. Acta Tropica 47, 251–418.CrossRefGoogle Scholar
Bird, A. F., Bonig, I. & Bacic, A. (1989). Factors affecting the adhesion of micro-organisms to the surfaces of plant-parasitic nematodes. Parasitology 98, 155–64.CrossRefGoogle Scholar
Bone, L. W. & Bottjer, K. P. (1985). Cuticular carbohydrates of three nematode species and chemoreception by Trichostrongylus colubriformis. Journal of Parasitology 71, 235–8.CrossRefGoogle ScholarPubMed
Devaney, E. (1985). Lectin binding characteristics of Brugia pahangi microfilariae. Tropical Medicine and Parasitology 36, 25–8.Google ScholarPubMed
Dunphy, G. B. & Webster, J. M. (1987). Partially characterized components of the epicuticle of dauer juvenile Steinernema feltiae and their influence on hemocyte activity in Galleria mellonella. Journal of Parasitology 73, 584–8.CrossRefGoogle Scholar
Forrest, J. M. S. & Robertson, W. M. (1986). Characterization and localization of saccharides on the head region of four populations of the potato cyst nematode Globodera rostochiensis and G. pallida. Journal of Nematology 18, 23–6.Google ScholarPubMed
Jansson, H.-B., Jeyaprakash, A., Coles, G. C., Marban-Mendoza, N. & Zuckerman, B. M. (1986). Fluorescent and ferritin labelling of cuticle surface carbohydrates of Caenorhabditis elegans and Panagrellus redivivus. Journal of Nematology 18, 570–4.Google ScholarPubMed
Kennedy, M. W. (1991). Parasitic Nematodes–Antigens, Membranes and Genes. London: Taylor and Francis.Google Scholar
Kennedy, M. W., Foley, M., Kuo, Y.-M., Kusel, J. R. & Garland, P. B. (1987). Biophysical properties of the surface lipid of parasitic nematodes. Molecular and Biochemical Parasitology 22, 233–40.CrossRefGoogle ScholarPubMed
Khoo, K.-H., Maizels, R. M., Page, A. P., Taylor, G. W., Rendell, N. & Dell, A. (1991). Characterisation of nematode glycoproteins: the major O-glycans of Toxocara excretory secretory antigens are methylated trisaccharides. Glycobiology 1, 163–71.CrossRefGoogle ScholarPubMed
Kiefer, E., Rudin, W. & Hecker, H. (1989). Cytochemical demonstration of lectin binding-sites in the cuticle and tissues of Acanthocheilonema viteae (Filarioidea). Acta Tropica 46, 3–15.CrossRefGoogle ScholarPubMed
Lloyd, K. O. & Kabat, E. A. (1968). Immunochemical studies on blood groups, XLI. Proposed structures for the carbohydrate portions of blood group A, B, H, Lewisa, and Lewisb substances. Proceedings of the National Academy of Sciences, USA 61, 1470–7.CrossRefGoogle Scholar
Lustigman, S., Huima, T., Brotman, B., Miller, K. & Price, A. M. (1990). Onchocerca volvulus: biochemical and morphological characteristics of the surface of third- and fourth-stage larvae. Experimental Parasitology 71, 489–95.CrossRefGoogle ScholarPubMed
Maizels, R. M., De Savigny, D. & Ogilvie, B. M. (1984). Characterization of surface and excretory–secretory antigens of Toxocara canis infective larvae. Parasite Immunology 6, 23–37.CrossRefGoogle ScholarPubMed
Maizels, R. M., Kennedy, M. W., Meghji, M., Robertson, B. D. & Smith, H. V. (1987). Shared carbohydrate epitopes on distinct surface and secreted epitopes of the parasitic nematode Toxocara canis. Journal of Immunology 139, 207–14.CrossRefGoogle ScholarPubMed
Maizels, R. M. & Selkirk, M. E. (1988). Immunobiology of nematode antigens. In The Biology of Parasitism (ed. Englund, P. T. & Sher, A.) pp. 285–308. New York: Alan R. Liss Inc.Google Scholar
Maizels, R. M., Robertson, B. D., Blaxter, M. L. & Selkirk, M. E. (1991). Parasite Antigens, Parasite Genes. A Laboratory Manual for Molecular Parasitology. Cambridge: Cambridge University Press.Google Scholar
McClure, M. A. & Zuckerman, B. M. (1982). Localization of cuticular binding sites of Concanavalin A on Caenorhabditis elegans and Meloidogyne incognita. Journal of Nematology 14, 39–44.Google ScholarPubMed
Meghji, M. & Maizels, R. M. (1986). Biochemical properties of larval excretory–secretory (ES) glycoproteins of the parasitic nematode Toxocara canis. Molecular and Biochemical Parasitology 18, 155–70.CrossRefGoogle Scholar
Ortega-Pierres, G., Chayen, A., Clark, N. W. T. & Parkhouse, R. M. E. (1984). The occurrence of antibodies to hidden and exposed determinants of surface antigens of Trichinella spiralis. Parasitology 88, 359–69.CrossRefGoogle ScholarPubMed
Page, A. P., Richards, D. T., Lewis, J. W., Omar, H. M. & Maizels, R. M. (1991). Comparison of isolates and species of Toxocara and Toxascaris by biosynthetic labelling of somatic and ES proteins from infective larvae. Parasitology 105, 451–64.CrossRefGoogle Scholar
Parsons, J. C., Bowman, D. D. & Grieve, R. B. (1986). Tissue localization of excretory–secretory antigens of larval Toxocara canis in acute and chronic murine toxocariasis. American Journal of Tropical Medicine and Hygiene 35, 974–81.CrossRefGoogle ScholarPubMed
Politz, S. M. & Philipp, M. (1992). Caenorhabditis elegans as a model for parasitic nematodes: a focus on the cuticle. Parasitology Today 8, 6–12.CrossRefGoogle Scholar
Rudin, W. (1990). Comparison of the cuticular structure of parasitic nematodes recognized by immunocytochemical and lectin binding studies. Acta tropica 47, 255–68.CrossRefGoogle ScholarPubMed
Slot, J. W. & Geuze, H. J. (1985). A new method of preparing gold probes for multiple labelling cytochemistry. European Journal of Cell Biology 38, 87–93.Google ScholarPubMed
Smith, H. V. (1991). Immune evasion and immunopathology in Toxocara canis infection. In Parasitic Nematodes Antigens, Membranes and Genes (ed. Kennedy, M. W.), pp. 116–39. London: Taylor and Francis.Google Scholar
Smith, H. V., Quinn, R., Kusel, J. R. & Girdwood, R. W. A. (1981). The effect of temperature and antimetabolites on antibody binding to the outer surface of second stage Toxocara canis larvae. Molecular and Biochemical Parasitology 4, 183–93.CrossRefGoogle Scholar
Smith, H. V., Kusel, J. R. & Girdwood, R. W. A. (1983). The production of human A and B blood group like substances by in vitro maintained second stage Toxocara canis larvae: their presence on the outer larval surfaces and in their excretions/secretions. Clinical and Experimental Immunology 54, 625–33.Google Scholar
Spiegel, Y. & McClure, M. A. (1991). Stage-specific differences in lectin binding to the surface of Anguina tritici and Meloidogyne incognita. Journal of Nematology 23, 259–63.Google Scholar
Sprent, J. F. A. (1958). Observations on the development of Toxocara canis (Werner, 1782) in the dog. Parasitology 48, 184–210.CrossRefGoogle ScholarPubMed
Zuckerman, B. (1983). Hypotheses and possibilities of intervention in nematode chemoresponses. Journal of Nematology 15, 173–82.Google ScholarPubMed
Zuckerman, B. M. & Jansson, H.-B. (1984). Nematode chemotaxis and possible mechanisms of host/prey recognition. Annual Review of Phytopathology 22, 95–113.CrossRefGoogle Scholar