Hostname: page-component-78c5997874-s2hrs Total loading time: 0 Render date: 2024-10-30T13:46:42.105Z Has data issue: false hasContentIssue false

Characterization of surface proteins and glycoproteins on red blood cells from mice infected with haemosporidia: Babesia rodhaini infections of BALB/c mice

Published online by Cambridge University Press:  06 April 2009

R. J. Howard
Affiliation:
The Laboratory of Immunoparasitology, The Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital P.O., Victoria 3050, Australia
Patricia M. Smith
Affiliation:
The Laboratory of Immunoparasitology, The Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital P.O., Victoria 3050, Australia
G. F. Mitchell
Affiliation:
The Laboratory of Immunoparasitology, The Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital P.O., Victoria 3050, Australia

Summary

Infection of intact (nu/+) or hypothymic (nu/nu) BALB/c mice with the lethal intra-erythrocytic parasite, Babesia rodhaini, induced several changes in the surface proteins of red cells from infected blood. Lactoperoxidase-catalysed radio-iodination was used to compare the surface proteins on normal mouse erythrocytes and the total red cells from infected blood at different levels of parasitaemia. At very low parasitaemia, when only 2·5% of the red cells contained parasites, we observed significant changes in the profile of radio-iodinated proteins separated by SDS-polyacrylamide gel electrophoresis. These changes included the appearance of a group of high molecular weight proteins, and a protein with an apparent molecular weight (Mr) of 60 000, both of which were absent from normal erythrocytes. The major labelled band on the erythrocyte surface (Mr 92 000) also appeared to be less heavily labelled during infection. The magnitude of these differences in surface proteins increased as the parasitaemia rose, until the new bands dominated the radioactivity profile with blood of greater than 50% parasitaemia. Several control experiments established that the radio-iodinated proteins were surface molecules on intact cells and that artifactual proteolysis did not contribute to the observed differences. The results suggest that changes in the surface proteins occur on all red cells in the blood of infected mice. The results of labelling the surface glycoproteins by oxidation with periodate or galactose oxidase, followed by NaB3H4 reduction, have varied with the isolate of B. rodhaini. With the isolate currently in use, no significant differences were observed in the labelled surface glycoproteins of normal erythrocytes and red cells from infected blood of high parasitaemia, whereas an earlier isolate exhibited a marked decrease of glycoprotein labelling of both infected and uninfected red cells.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1980

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

Bonner, W. M. & Laskey, R. A. (1974). A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46, 83–8.CrossRefGoogle ScholarPubMed
Callow, L. L. (1977). Vaccination against bovine babesiosis. In Immunity to Blood Parasites of Animals and Man, (ed. Miller, L. H., Pino, J. A. and McKelvey, J. J. Jr), pp. 121–49. New York: Plenum Publishing Corporation.CrossRefGoogle Scholar
Chapman, W. D. & Ward, P. A. (1977). Babesia rodhaini: requirement of complement for penetration of human erythrocytes. Science 196, 67–9.CrossRefGoogle ScholarPubMed
Cox, K. O., Howard, R. J. & Mitchell, G. F. (1977). Studies on immune responses to parasite antigens in mice. VII. Cells secreting antibodies to modified mouse erythrocytes in Babesia rodhaini-infected mice. Cellular Immunology 32, 223–7.CrossRefGoogle Scholar
Crumpton, M. J., Marchalonis, J. J., Haustein, D., Atwell, J. L. & Harris, A. W. (1976). Plasma membrane of a murine T cell lymphoma: surface labelling, membrane isolation, separation of membrane proteins and distribution of surface label amongst these proteins. Australian Journal of Experimental Biology and Medical Science 54, 303–16.CrossRefGoogle ScholarPubMed
Crumpton, M. J. & Snary, D. (1974). Preparation and properties of lymphocyte plasma membrane. In Contemporary Topics in Molecular Immunology, vol. 3 (ed. Ada, G. L.), pp. 2756. New York: Plenum Publishing Corporation.CrossRefGoogle Scholar
Curnow, J. A. (1968). In vitro agglutination of bovine erythrocytes infected with Babesia argentina. Nature, London 217, 267–8.CrossRefGoogle ScholarPubMed
Curnow, J. A. (1973). Studies on antigenic changes and strain differences in Babesia argentina infections. Australian Veterinary Journal 49, 279–83.CrossRefGoogle ScholarPubMed
Dodge, J. T., Mitchell, C. & Hanahan, D. J. (1963). The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Archives of Biochemistry and Biophysics 100, 119–30.CrossRefGoogle ScholarPubMed
Fairbanks, G., Steck, T. L. & Wallach, D. F. H. (1971). Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10, 2606–17.CrossRefGoogle ScholarPubMed
Gahmberg, C. G. (1976). External labeling of human erythrocyte glycoproteins. Studies with galactose oxidase and fluorography. Journal of Biological Chemistry 251, 510–15.CrossRefGoogle ScholarPubMed
Gahmberg, C. G. & Hakomori, S. (1973). External labeling of cell surface galactose and galactosamine in glycolipid and glycoprotein of human erythrocytes. Journal of Biological Chemistry 248, 4311–17.CrossRefGoogle ScholarPubMed
Gahmberg, C. G., Hayry, P. & Andersson, L. C. (1976). Characterization of surface glycoproteins of mouse lymphoid cells. Journal of Cell Biology 68, 642–53.CrossRefGoogle ScholarPubMed
Howard, R. J. & Rodwell, B. (1979). Babesia rodhaini, Babesia bovis and Babesia bigemina: analysis and sorting of red cells from infected mouse or calf blood by flow fluorimetry using 33258 Hoechst. Experimental Parasitology 48, 421–31.CrossRefGoogle ScholarPubMed
Howard, R. J., Smith, P. M. & Mitchell, G. F. (1978). Removal of leucocytes from red cells in Plasmodium berghei-infected mouse blood and purification of schizont-infected cells. Annals of Tropical Medicine and Parasitology 72, 573–6.CrossRefGoogle ScholarPubMed
Howard, R. J., Smith, P. M. & Mitchell, G. F. (1979 a). Identification of several differences between external proteins and glycoproteins of normal mouse (BALB/c) and human erythrocytes. Journal of Membrane Biology 49, 171–98.CrossRefGoogle ScholarPubMed
Howard, R. J., Smith, P. M. & Mitchell, G. F. (1979 b). Optimal conditions for lactoperoxidase catalyzed radioiodination of external proteins on mouse erythrocytes. Australian Journal of Experimental Biology and Medical Science 57, 355–68.CrossRefGoogle ScholarPubMed
Howard, R. J., Smith, P. M. & Mitchell, G. F. (1980 a). Characterization of surface proteins and glycoproteins on red blood cells from mice infected with haemosporidia. Plasmodium berghei infectious of BALB/c mice. Parasitology 81, 273–98.CrossRefGoogle Scholar
Howard, R. J., Smith, P. M. & Mitchell, G. F. (1980 b). Characterization of surface proteins and glycoproteins on red blood cells from mice infected with haemosporidia. Plasmodium yoelii infections of BALB/c mice. Parasitology 81, 299314.CrossRefGoogle ScholarPubMed
Hubbard, A. L. & Cohn, Z. A. (1972). The enzymatic iodination of the red cell membrane. Journal of Cell Biology 55, 390405.CrossRefGoogle ScholarPubMed
Hynes, R. O. (1977). Surface labelling techniques for eukaryotic cells. In New Techniques in Biophysical and Cell Biology, vol. 3, (ed. Pain, R. and Smith, B. J.), pp. 147212. Wiley Int. Publ.Google Scholar
Laemmli, U. K. & Favre, M. (1973). Maturation of the head of bacteriophage T4.I.DNA packaging events. Journal of Molecular Biology 80, 575–99.CrossRefGoogle ScholarPubMed
Mahoney, D. F. (1978). Babesiosis of domestic animals. In Parasitic Protozoa, vol. 4 (ed. Kreier, J. P.), pp. 152. New York: Academic Press.Google Scholar
Mitchell, G. F. (1977). Studies on immune responses to parasite antigens in mice. V. Different susceptibilities of hypothymic and intact mice to Babesia rodhaini. International Archives of Allergy and Applied Immunology 53, 385–8.CrossRefGoogle ScholarPubMed
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 larvae numbers and antiphosphorylcholine responses in infected mice of various strains and in hypothymic nu/nu mice. International Archives of Allergy and Applied Immunology 52, 6478.CrossRefGoogle ScholarPubMed
Moore, G. L., Kocholaty, W. F., Cooper, D. A., Gray, J. L. & Robinson, S. L. (1970). A proteinase from human erythrocyte membranes. Biochimica et biophysica acta 212, 126–33.CrossRefGoogle ScholarPubMed
Nomoto, M., Narahashi, Y. & Murakami, M. (1960 a). A proteolytic enzyme of Streptomyces griseus. VI. Hydrolysis of protein by Streptomyces griseus protease. Journal of Biochemistry, Tokyo 48, 593602.CrossRefGoogle Scholar
Nomoto, M., Narahashi, Y. & Murakami, M. (1960 b). A proteolytic enzyme of Streptomyces griseus VII. Substrate specificity of Streptomyces griseus protease. Journal of Biochemistry, Tokyo 48, 906–18.CrossRefGoogle Scholar
Roberts, J. A. & Tracey-Patte, P. (1975). Babesia rodhaini: immunoinduction of antigenic variation. International Journal for Parasitology 5, 573–6.CrossRefGoogle ScholarPubMed
Schroeder, W. F., Cox, H. W. & Ristic, M. (1966). Anaemia, parasitaemia, erythrophagocytosis, and haemagglutinins in Babesia rodhaini infection. Annals of Tropical Medicine and Parasitology 60, 31–8.CrossRefGoogle ScholarPubMed
Sibinovic, K. H., Milar, R., Ristic, M. & Cox, H. W. (1969). In vivo and in vitro effects of serum antigens of babesial infection and their antibodies on parasitized and normal erythrocytes. Annals of Tropical Medicine and Parasitology 63, 327–36.CrossRefGoogle ScholarPubMed
Steck, T. L. & Dawson, G. (1974). Topographical distribution of complex carbohydrates in the erythrocyte membrane. Journal of Biological Chemistry 249, 2135–42.CrossRefGoogle ScholarPubMed
Swanstrom, R. & Shank, P. R. (1978). X-ray intensifying screens greatly enhance the detection by autoradiography of the radioactive isotopes 32P and 125I. Analytical Biochemistry 86, 184–92.CrossRefGoogle ScholarPubMed
Tokes, Z. A. & Chambers, S. M. (1975). Proteolytic activity associated with human erythrocyte membranes. Self-digestion of isolated human erythrocyte membranes. Biochimica et biophysica acta 389, 325–8.CrossRefGoogle ScholarPubMed
Trop, M. & Birk, Y. (1970). The specificity of proteinases from Streptomyces griseus (Pronase). The Biochemical Journal 116, 1925.CrossRefGoogle ScholarPubMed
Wright, B. M., Edwards, A. J. & Jones, V. E. (1974). Use of a cell rupturing pump for the preparation of thymocyte subcellular fractions. Journal of Immunological Methods 4, 281–96.CrossRefGoogle ScholarPubMed
Wright, I. G. (1973 a). Observations on the haematology of experimentally induced Babesia argentina and B. bigemina infections in splenectomized calves. Research in Veterinary Science 17, 2934.CrossRefGoogle Scholar
Wright, I. G. (1973 b). Plasma kallikrein levels in acute Babesia argentina infections in splenectomized and intact calves. Zeitschrift für Parasitenkunde 41, 269–80.CrossRefGoogle Scholar
Wright, I. G. (1975). The probable role of Babesia argentina esterase in the in vitro activation of plasma prekallikrein. Veterinary Parasitology 1, 91–6.CrossRefGoogle Scholar
Wright, I. G. & Goodger, B. V. (1973). Proteolytic enzyme activity in the intraerythrocytic parasites Babesia argentina and Babesia bigemina. Zeitschrift für Parasitenkunde 42, 213–20.Google ScholarPubMed
Wright, I. G. & Kerr, J. D. (1975). Effect of trasylol on packed cell volume and plasma kallikrein activation in acute Babesia argentina infection of splenectomized calves. Zeitschrift für Parasitenkunde 46, 189–94.CrossRefGoogle Scholar
Wright, I. G. & Mahoney, D. F. (1974). The activation of kallikrein in acute Babesia argentina infections in splenectomized calves. Zeitschrift für Parasitenkunde 43, 271–8.CrossRefGoogle Scholar