Hostname: page-component-78c5997874-t5tsf Total loading time: 0 Render date: 2024-11-19T22:18:43.139Z Has data issue: false hasContentIssue false
  • English
  • Français

Murine hybridoma-derived antibodies in the processing of antigens for the immunodiagnosis of hydatid (Echinococcus granulosus) infection in sheep

Published online by Cambridge University Press:  08 November 2011

P. S. Craig ,
R. E. Hocking ,
G. F. Mitchell  and
M. D. Rickard
P. S. Craig
Affiliation:
Veterinary Clinical Centre, University of Melbourne, Princes Highway, Werribee, Victoria 3030, Australia
R. E. Hocking
Affiliation:
Laboratory of Immunoparasitology, The Walter and Eliza Hall Institute of Medical Research, Melbourne 3050, Victoria, Australia
G. F. Mitchell
Affiliation:
Veterinary Clinical Centre, University of Melbourne, Princes Highway, Werribee, Victoria 3030, Australia
M. D. Rickard
Affiliation:
Veterinary Clinical Centre, University of Melbourne, Princes Highway, Werribee, Victoria 3030, Australia
Get access Rights & Permissions [Opens in a new window]

Summary

Mice experimentally infected with the hydatid parasite Echinococcus granulosus (secondary echinococcosis), were used as spleen cell donors for hybridoma production. Two hybridomas were produced which secreted antibody with anti-E. granulosus cyst fluid (EgCF) activity. Radio-immunoassays comparing the binding of these hybridoma-derived antibodies to E. granulosus versus several other antigen preparations from sheep parasites showed that the antibodies had a high degree of specificity for the hydatid parasite. However, using a panel of clinically defined sheep sera in a competitive radio-immunoassay, the binding of the hybridoma-derived antibodies to EgCF was inhibited strongly by sera from sheep infected with the common parasites Fasciola hepatica and Taenia hydatigena. When the hybridoma-derived antibodies were conjugated to CNBr-activated Sepharose and the crude EgCF applied to the affinity column, the ‘run through’ antigen preparation showed a significant increase in specificity for experimental E. granulosus infection. A useful means of applying the hybridoma technology in the development of new immunodiagnostic reagents may be to select those murine hybridomas which produce high-affinity, cross-reacting antibodies and to use these antibodies in the processing of homologous antigen by affinity chromatography.

Type
Research Article
Information
Parasitology , Volume 83 , Issue 2 , October 1981 , pp. 303 - 317
Copyright
Copyright © Cambridge University Press 1981

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

Araj, G. F., Matossian, R. M. & Frayha, G. J. (1977). The host response in secondary hydatidosis of mice. I. Circulating antibodies. Zeitschrift für Parasitenkunde 52, 23–30.CrossRefGoogle ScholarPubMed
Boyd, A. W. & Schrader, J. W. (1980). Mechanism of effector-cell blockade. I. Antigen-induced suppression of Ig synthesis in a hybridoma cell line, and correlation with cell-associated antigen. Journal of Experimental Medicine 151, 14–36.CrossRefGoogle Scholar
Capron, A., Vernes, A. & Biguet, J. (1967). Le diagnostic immunoelectrophorétique de l'hydatidose. In 8IMEP Ed. (Lyon). Le kyste hydatique du foie (Journées Lyonnaises d'Hyda-tidologie, 1966), pp. 27–40.Google Scholar
Craig, P. S., Mitchell, G. F., Cruise, K. M. & Rickard, M. D. (1980). Hybridoma antibody immunoassay for the detection of parasitic infection: Attempts to produce an immunodiag-nostic reagent for larval taeniid cestode infection. Australian Journal of Experimental Biology and Medical Science 58, 339–50.CrossRefGoogle ScholarPubMed
Craig, P. S. & Rickard, M. D. (1981). Anti-oncospheral antibodies in the serum of lambs experimentally infected with either Taenia ovis or Taenia hydatigena. Zeitschrift für Parasitenkunde 64, 169–77.CrossRefGoogle ScholarPubMed
Craig, P. S. & Rickard, M. D. (1980). Evaluation of ‘crude’ antigen prepared from Taenia saginata for the serological diagnosis of T. saginata cysticercosis in cattle using the enzyme-linked immunosorbent assay (ELISA). Zeitschrift für Parasite.nkunde. 61, 287–97.CrossRefGoogle ScholarPubMed
Galfre, G., Howe, S. C., Milstein, C., Butcher, G. W. & Howard, J. C. (1977), Antibodies to major histocompatability antigens produced by hybrid cell lines. Nature, London 266, 546–52.CrossRefGoogle ScholarPubMed
Hämmerling, G. J. (1977). T. lymphocyte tissue culture lines produced by cell hybridization. European Journal of Immunology 7, 743—5.CrossRefGoogle ScholarPubMed
Hartree, F. E. (1972). Determination of protein: a modification of the Lowry method that gives a linear photometric response. Analytical Biochemistry 48, 422–7.CrossRefGoogle ScholarPubMed
Heath, D. D. (1970). The development of Echinococcus granulosus larvae in laboratory animals. Parasitology 60, 449–56.CrossRefGoogle ScholarPubMed
Kagan, I. G. (1968). A review of serological tests for the diagnosis of hydatid disease. Bulletin of the World Health Organization 39, 25–37.Google ScholarPubMed
Köhler, G. & Milstein, C. (1975). Continuous cultures of fused cells secreting antibody of predefined specificity. Nature, London 256, 495–7.CrossRefGoogle ScholarPubMed
Köhler, G. & Milstein, C. (1976). Derivation of specific antibody-producing tissue culture and tumour lines by cell fusion. European Journal of Immunology 6, 511–19.CrossRefGoogle ScholarPubMed
Marcoullis, G., Salonen, E. M. & Grasbeck, R. (1978). Sequential affinity chromatography for the purification of antigens extracted from Onchocerca volvulus adult worms. Zeitschrift für Tropenmedizin und Parasitologie 29, 39–48.Google ScholarPubMed
Matossian, R. M. (1977). The immunological diagnosis of human hydatid disease. Transactions of the Royal Society of Tropical Medicine and Hygiene 71, 101–4.CrossRefGoogle ScholarPubMed
Matossian, R. M., Rickard, M. D. & Smyth, J. D. (1977). Hydatidosis: a global problem of increasing importance. Bulletin of the World Health Organization 55, 499–507.Google ScholarPubMed
Mitchell, G. F., Cruise, K. M., Chapman, C. B., Anders, R. F. & Howard, M. C (1979). Hybridoma antibody immunoassays for the detection of parasitic infection: development of a model system using a larval cestode infection in mice. Australian Journal of Experimental Biology and Medical Science 57, 287–302.CrossRefGoogle Scholar
Mitchell, G. F., Hogarth-Scott, R. S., Edwards, R. D., Lewers, H. M., Cousins, G. & Moore, T. (1976). Studies on immune responses to parasite antigens in mice. I. Ascaris suum larval numbers and antiphosphorylcholine responses in mice of various strains and in hypothymic nude mice. International Archives of Allergy and Applied Immunology 52, 64–78.CrossRefGoogle Scholar
Pauluzzi, S. (1969). Serological response of mice and rats to secondary experimental hydatid disease. American Journal of Tropical Medicine and Hygiene 18, 7–12.CrossRefGoogle ScholarPubMed
Rickard, M. D. (1979). The immunological diagnosis of hydatid disease. Australian Veterinary Journal 55, 99–104.CrossRefGoogle Scholar
Smyth, J. D. (1962). Studies on tapeworm physiology. X. Axenic cultivation of the hydatid organism, Echinococcus granulosus; establishment of a basic technique. Parasitology 52, 441–57.CrossRefGoogle Scholar
Suzuki, T., Sato, Y., Yamashita, T., Sekikawa, H. & Otsuru, M. (1975). Angiostrongylus cantonensis: Preparation of a specific antigen using immunoadsorbent columns. Experimental Parasitology 38, 191–201.CrossRefGoogle ScholarPubMed
Varela-Diaz, V. M., Coltorti, E. A., Rickard, M. D. & Torres, J. M. (1977). Comparative antigenic characterization of Echinococcus granulosus and Taenia hydatigena cyst fluids by immunoelectrophoresis. Research in Veterinary Science 23, 213–16.CrossRefGoogle ScholarPubMed
Welch, J. W. & Dobson, C. (1978). Immunodiagnosis of parasitic zoonoses: Comparative efficacy of three immunofluorescence tests using antigens purified by affinity chromatography. Transactions of the Royal Society for Tropical Medicine and Hygiene 72, 282–8.CrossRefGoogle ScholarPubMed
Yong, W. K. & Heath, D. D. (1979). ‘Arc 5’ antibodies in sera of sheep infected with Echinococcus granulosus, Taenia hydatigena and Taenia ovis. Parasite Immunology 1, 27–38.CrossRefGoogle ScholarPubMed