Hostname: page-component-586b7cd67f-gb8f7 Total loading time: 0 Render date: 2024-11-28T07:25:00.048Z Has data issue: false hasContentIssue false
  • English
  • Français

Echinococcus granulosus coproantigens: chromatographic fractionation and characterization

Published online by Cambridge University Press:  16 April 2004

F. A. ELAYOUBI  and
P. S. CRAIG
F. A. ELAYOUBI
Affiliation:
Cestode Zoonoses Research Group, Biosciences Research Institute, School of Environment and Life Sciences, University of Salford, Salford M5 4WT, UK
P. S. CRAIG
Affiliation:
Cestode Zoonoses Research Group, Biosciences Research Institute, School of Environment and Life Sciences, University of Salford, Salford M5 4WT, UK
Get access Rights & Permissions [Opens in a new window]

Abstract

Dogs infected with adult tapeworms of Echinococcus granulosus release antigens (coproantigens) in faeces which can be detected by a capture ELISA. Supernatants prepared from E. granulosus-infected dog faecal samples were fractionated by size-exclusion fast protein liquid chromatography (FPLC) on a Superose-6 column. Coproantigen ELISA and Western blotting were used to demonstrate the immunoreactivity of eluted fractions. Two main FPLC peaks of antigenic activity were detected and designated as fraction F1 and fraction F2 with approximate relative molecular weights >670 kDa, and in the range of 146 to 440 kDa respectively. These two antigenic fractions (F1 and F2) fractionated from infected dog faeces were heat stable and largely protease-insensitive, but were highly sensitive to sodium periodate treatment, which strongly suggested the involvement of carbohydrates. Capture IgG antibodies against E. granulosus proglottis somatic extracts, detected a molecule with an approximate molecular weight of 155 kDa in fraction F2 after immunoblotting. The 155 kDa antigen could be completely ablated by sodium periodate treatment, but not after protease or lipase treatment. A surface tegument preparation of adult E. granulosus tapeworms contained large amounts of antigen that corresponded in size range and antigenicity to that observed in the FPLC fraction F2. There was also a peak of antigenic activity at >670 kDa corresponding to fraction F1 from a culture derived excretory–secretory (E–S) adult tapeworm preparation. The involvement of carbohydrate moieties in coproantigen activity present in the FPLC fractions F1 and F2 from faecal supernatants of E. granulosus-infected dogs was confirmed by lectin-binding assays and exoglycosidase treatment, which showed that α-D-mannose and/or α-D-glucose, β-galactose and N-acetyl-β-glucosamine residues were the most important carbohydrate components in putative coproantigens present in both fractions. N-acetyl-β-glucosamine and sialic acid residues were also contained in coproantigen molecules present in fraction F2. These results suggested that coproantigens detected in faeces of E. granulosus-infected dogs are large molecular weight molecules that may be derived from the carbohydrate-rich surface glycocalyx of adult worms, and are shed, released or secreted during the life-span of the tapeworm.

Keywords

Echinococcus granulosuscoproantigenFPLCperiodatelectinexoglycosidase
Type
Research Article
Information
Parasitology , Volume 128 , Issue 4 , April 2004 , pp. 455 - 465
Copyright
© 2004 Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

ALLAN, J. C. & CRAIG, P. S. (1994). Partial characterization and time course analysis of Hymenolepis diminuta coproantigens. Journal of Helminthology 68, 97–103.CrossRefGoogle Scholar
ALLAN, J. C., CRAIG, P. S., GARCIA NOVAL, J., MENCOS, F., LIU, D., WANG, Y., WEN, H., ZHOU, P., STRINGER, R., ROGAN, M. & ZEYHLE, E. (1992). Coproantigen detection for immunodiagnosis of echinococcosis and taeniasis in dogs and humans. Parasitology 104, 347–355.CrossRefGoogle Scholar
BERRADA-RKHAMI, O., LEDUCQ, R., GABRION, J. & GABRION, C. (1990). Selective distribution of sugars on the tegumental surface of adult Bothriocephalus gregarius (Cestoda: Pseudophyllidea). International Journal for Parasitiology 20, 285–297.CrossRefGoogle Scholar
CASARAVILLA, C., MALGOR, R. & CARMONA, C. (2003). Characterization of carbohydrates of adult Echinococcus granulosus by lectin-binding analysis. Journal of Parasitology 89, 57–61.CrossRefGoogle Scholar
CRAIG, P. S. (1997). Immunodiagnosis of Echinococcus granulosus and a comparison of techniques for diagnosis of canine echinococcosis. In Compendium of Cystic Echinococcosis in Africa and in the Middle Eastern Countries with Special Reference to Morocco (ed. Andersen, F. L., Ouhelli, H. & Kachani, M. ), pp. 85–118. Brigham Young University Print Services, Provo, Utah, USA.
DEPLAZES, P., GOTTSTEIN, B., ECKERT, J., JENKINS, D. J., EWALD, D. & JIMENEZ-PALACIOS, S. (1992). Detection of Echinococcus coproantigens by Enzyme-Linked Immunosorbent Assay in dogs, dingoes and foxes. Parasitology Research 78, 303–308.CrossRefGoogle Scholar
DEPLAZES, P., ALTHER, P., TANNER, I., THOMPSON, R. C. A. & ECKERT, J. (1999). Echinococcus multilocularis coproantigen detection by enzyme-linked immunosorbent assay in fox, dog, and cat populations. Journal of Parasitology 85, 115–121.CrossRefGoogle Scholar
ELAYOUBI, F. A., FRASER, A., JENKINS, D. J. & CRAIG, P. S. (2003). Partial characterisation of carbohydrate-rich Echinococcus granulosus coproantigens. International Journal for Parasitology 33, 1553–1559.CrossRefGoogle Scholar
EL-SHEHABI, F. S., KAMHAWI, S. A., SCHANTZ, P. M., CRAIG, P. S. & ABDEL-HAFEZ, S. K. (2000). Diagnosis of canine echinococcosis: Comparison of coproantigen detection with necropsy in stray dogs and red foxes from Northern Jordan. Parasite 7, 83–90.CrossRefGoogle Scholar
FRASER, A. & CRAIG, P. S. (1997). Detection of gastrointestinal helminth infections using coproantigen and molecular diagnostic approaches. Journal of Helminthology 71, 103–107.CrossRefGoogle Scholar
FRIEDMAN, P. A., WEINSTEIN, P. P., DAVIDSON, L. A. & MUELLER, J. F. (1982). Spirometra mansonoides: lectin analysis of tegumental glycopeptides. Experimental Parasitology 54, 93–103.CrossRefGoogle Scholar
GUARNERA, E. A., SANTILLAN, G., BOTINELLI, R. & FRANCO, A. (2000). Canine echinococcosis: an alternative for surveillance epidemiology. Veterinary Parasitology 88, 131–134.CrossRefGoogle Scholar
ITO, A. & SMYTH, J. D. (1987). Adult cestodes – immunology of the lumen-dwelling cestode infections. In Immune Response in Parasite Infections: Immunology, Immunopathology and Immunoprophylaxis. Vol. II: Trematodes and Cestodes (ed. Soulsby, E. J. L. ), pp. 115–163. CRC Press, Boca Raton, USA.
JENKINS, D. J., FRASER, A., BRADSHAW, H. & CRAIG, P. S. (2000). Detection of Echinococcus granulosus coproantigens in Australian canids with natural or experimental infection. Journal of Parasitology 86, 140–145.CrossRefGoogle Scholar
KOHNO, H., SAKAI, H., OKAMOTO, M., ITO, M., OKU, Y. & KAMIYA, M. (1995). Development and characterization of murine monoclonal antibodies to Echinococcus multilocularis adult worms and its use for the coproantigen detection. Japanese Journal of Parasitology 44, 404–412.Google Scholar
LAEMMLI, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–685.CrossRefGoogle Scholar
LUMSDEN, R. D. (1975). Surface ultrastructure and cytochemistry of parasitic helminths. Experimental Parasitology 37, 267–339.CrossRefGoogle Scholar
LUMSDEN, R. D., OAKS, J. A. & ALWORTH, W. L. (1970). Cytological studies on the absorptive surface of cestodes. IV. Localization and cytochemical properties of membrane-fixed cation binding sites. Journal of Parasitology 56, 736–747.Google Scholar
MAASS, M., DELGADO, E. & KNOBLOCH, J. (1992). Isolation of an immunodiagnostic Taenia solium coproantigen. Tropical Medicine of Parasitology 43, 201–202.Google Scholar
MACHNICKA, B., DZIEMIAN, E. & ZWIERZ, C. (1996). Factors conditioning detection of Taenia saginata antigens in faeces. Applied Parasitology 37, 99–105.Google Scholar
MALGOR, R., NONAKA, N., BASMADJIAN, I., SAKAI, H., CARAMBULA, B., OKU, Y., CARMONA, C. & KAMIYA, M. (1997). Coproantigen detection in dogs experimentally and naturally infected with Echinococcus granulosus by a monoclonal antibody-based Enzyme-Linked Immunosorbent Assay. International Journal for Parasitology 27, 1605–1612.CrossRefGoogle Scholar
NONAKA, N., IIDA, M., YAGI, K., ITO, T., OOI, H. K., OKU, Y. & KAMIYA, M. (1996). Time course of coproantigen excretion in Echinococcus multilocularis infections in foxes, and alternative definitive host, golden hamsters. International Journal for Parasitology 26, 1271–1278.CrossRefGoogle Scholar
OSINAGA, E., PANCINO, G., PORCHET, N., BEROIS, N., DE CREMOUX, P., MISTRO, D., AUBERT, J. P., CALVO, F. & ROSETO, A. (1994). Analysis of a heterogeneous group of human breast carcinoma associated glycoproteins bearing the Tn determinant. Breast Cancer Research and Treatment 32, 139–152.CrossRefGoogle Scholar
SAKAI, H., MALGOR, R., BASMADJIAN, I., GALLARDO, R., CARMONA, C., SATO, H., OKU, Y. & KAMIYA, M. (1995). An Enzyme-Linked Immunosorbent Assay (ELISA) for detection of Echinococcus granulosus coproantigens in dogs. Japanese Journal of Parasitology 44, 453–461.Google Scholar
SAKASHITA, M., SAKAI, H., KOHNO, H., OOI, H., OKU, Y., YAGI, K., ITO, M. & KAMIYA, M. (1995). Detection of Echinococcus multilocularis coproantigens in experimentally infected dogs using murine monoclonal antibody against adult worms. Japanese Journal of Parasitology 44, 413–420.Google Scholar
SANDEMAN, R. M. & WILLIAMS, J. F. (1984). Lectin binding to cystic stages of Taenia taeniaeformis. Journal of Parasitology 70, 661–667.CrossRefGoogle Scholar
SCHMIDT, J. (1988). Expression of glycoconjugates on normally developing and immunologically impaired Hymenolepis diminuta. Parasitology Research 75, 155–161.CrossRefGoogle Scholar
SCHMIDT, J. & PETERS, W. (1987). Localization of glycoconjugates at the tegument of the tapeworms Hymenolepis nana and H. microstoma with gold labelled lectins. Parasitology Research 73, 80–86.Google Scholar
TOWBIN, H., STAEHELIN, T. & GORDON, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. Proceedings of the National Academy of Sciences, USA 76, 4350–4354.CrossRefGoogle Scholar
WHITFIELD, P. J. (1979). The Biology of Parasitism. Arnold, London.
WHO/OIE (2001). Manual on Echinococcosis in Human and Amimals: a Public Health Problem of Global Concern ( ed. Eckert, J., Gemmell, M. A., Meslin, F. X. & Pawlowski, Z. S.). World Organization for Animal Health, France.