Hostname: page-component-78c5997874-s2hrs Total loading time: 0 Render date: 2024-11-04T19:10:28.459Z Has data issue: false hasContentIssue false
  • English
  • Français

A monoclonal antibody-derived antigen of Babesia divergens: characterization and investigation of its ability to protect gerbils against virulent homologous challenge

Published online by Cambridge University Press:  06 April 2009

C. M. Winger ,
E. U. Canning  and
J. D. Culverhouse
C. M. Winger
Affiliation:
Department of Pure and Applied Biology, Imperial College of Science, Technology and Medicine, London SW1 2AZ
E. U. Canning
Affiliation:
Department of Pure and Applied Biology, Imperial College of Science, Technology and Medicine, London SW1 2AZ
J. D. Culverhouse
Affiliation:
Department of Pure and Applied Biology, Imperial College of Science, Technology and Medicine, London SW1 2AZ
Get access Rights & Permissions [Opens in a new window]

Summary

A Babesia divergens merozoite antigen was purified by affinity chromatography using a monoclonal antibody. Silver staining of SDS-PAGE gels revealed 2 bands of Mr 50–60 kDa and Mr 24–29. It is proposed that the Mr 24–29 kDa band represents the native protein and that it is complexed, perhaps as a dimer or perhaps to host protein to give the Mr 50–60 kDa band. Peptide analysis of the antigen followed by Western blotting with the monoclonal antibody revealed only an Mr 20–24 kDa band. After affinity purification the antigen was precipitated with acetone and inoculated subcutaneously, together with complete Freund's adjuvant, into gerbils. Two inoculations were given 3 weeks apart, after which the immunized animals and a group of control animals were challenged with 5 × parasitized red cells from a virulent microaerophilus stationary phase (MASP) culture of an homologous strain of B. divergens. Parasitaemias rose rapidly in control animals and all were dead by the fifth day. Two of the vaccinated animals survived the challenge. Parasitaemias in the remaining two rose slowly but both animals succumbed on the seventh day. Western blotting of merozoites and of the purified antigen, using sera from the surviving animals showed strong reactivity with the Mr 24–29 kDa band.

Keywords

Babesia divergensantigencharacterizationprotectionmerozoite.
Type
Research Article
Information
Parasitology , Volume 99 , Issue 3 , December 1989 , pp. 341 - 348
Copyright
Copyright © Cambridge University Press 1989

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

Gooderham, K. (1984). In situ peptide mapping of proteins following polyacrylamide gel electrophoresis. In Methods in Molecular Biology, Vol. 1. ‘Proteins’, (ed. Walker, J. M.), Clifton, N.J.: Humana Press. pp. 192–202.Google Scholar
Köhler, G. & Milstein, C. (1975). Continuous cultures of fused cells secreting antibody of predefined specificity. Nature, London 256, 495–7.CrossRefGoogle ScholarPubMed
Laemmli, u. K. (1970). Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature, London 277, 680–5.Google Scholar
Levy, M. G. & Ristic, M. (1980). Babesia bovis: continuous cultivation in a microaerophilous stationary phase culture. Science 207, 1218–20.CrossRefGoogle Scholar
Mcelwain, T. F., Perryman, L. E., Davis, W. C. & Mcguire, T. C. (1987). Antibodies define multiple proteins with epitopes exposed on the surface of live Babesia bigemina merozoites. Journal of Immunology 138, 2298–304.CrossRefGoogle ScholarPubMed
Ristic, M. (1988). Exoantigens of Babesia. In Babesiosis of Domestic Animals and Man (ed. Ristic, M.), pp. 264.Google Scholar
TAYLOR, S. M., ELLIOTT, C. T. & KENNY, T. (1986). Isolation of antigenic proteins from erythrocytes parasitised with Babesia divergens and a comparison of their immunising potential. Veterinary Parasitology 21, 99–105.CrossRefGoogle Scholar
Taylor, S. M., Kenny, J., Mallon, T. & Elliott, C. T. (1984). The immunisation of cattle against Babesia divergens with fractions of parasitised erythrocytes. Veterinary Parasitology 16, 235–42.CrossRefGoogle Scholar
Voller, A., Bidwell, D. E. & Bartlett, A. (1976). Enzyme immunoassays in diagnostic medicine. Theory and practice. Bulletin of the World Health Organization 53, 3418–21.Google Scholar
Winger, C. M., Canning, E. U. & Culverhouse, J. D. (1987 a). A monoclonal antibody to Babesia divergens which inhibits merozoite invasion. Parasitology 94, 17–27.CrossRefGoogle ScholarPubMed
Winger, C. M., Canning, E. U. & Culverhouse, J. D. (1987 b). Induction of protective immunity to Babesia divergens in Mongolian gerbils, Meriones unguiculatus, using culture-derived immunogens. Veterinary Parasitology 26, 43–53.Google Scholar
Winger, C. M., Canning, E. U. & Culverhouse, J. D. (1989). A strain of Babesia divergens attenuated after long term culture. Research in Veterinary Science 46, 110–13.Google Scholar
Wright, I. G., Mirre, G. B., Rode-Bramanis, K., Chamberlain, M., Goodger, B. V. & Waltisbuhl, D. J. (1985). Protective vaccination against virulent Babesia bovis with a low-molecular-weight antigen. Infection and Immunity 48, 109–13.CrossRefGoogle ScholarPubMed
Wright, I. C., White, M., Tracey-Patte, P. D., Donaldson, R. A., Goodger, B. V., Waltisbuhl, D.J. & Mahoney, D. F. (1983). Babesia bovis: isolation of a protective antigen by using monoclonal antibodies. Infection and Immunity 41, 244–50.CrossRefGoogle ScholarPubMed