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Theileria annulata-infected cells produce abundant proteases whose activity is reduced by long-term cell culture

Published online by Cambridge University Press:  06 April 2009

H. A. Baylis ,
A. Megson ,
C. G. D. Brown ,
G. F. Wilkie  and
R. Hall
H. A. Baylis
Affiliation:
Department of Biology, University of York, Heslington, York YO1 5DD, UK
A. Megson
Affiliation:
Department of Biology, University of York, Heslington, York YO1 5DD, UK
C. G. D. Brown
Affiliation:
Centre for Tropical Veterinary Medicine, University of Edinburgh, Roslin, UK
G. F. Wilkie
Affiliation:
Centre for Tropical Veterinary Medicine, University of Edinburgh, Roslin, UK
R. Hall
Affiliation:
Department of Biology, University of York, Heslington, York YO1 5DD, UK
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Summary

Lysates of Theileria annulata-infected bovine lymphoblastoid cells and their uninfected counterparts were tested for protease activity using gelatin substrate SDS–PAGE. The infected cells produced a number of extra activities at pH 8·0 in the presence of Ca2+. Calcium was found to enhance the activites but was not an absolute requirement. Studies using inhibitors, including E64, 3,4-dichloroisocoumarin, pepstatin and 1, 10-phenanthroline suggested that the activities were metalloproteases. We analysed two vaccine lines; the Ode line from India and the Ankara Pendik line from Turkey. In the Ode line the later passage had very much reduced levels of the enzyme activities. In the case of the Ankara Pendik line both stages analysed had very low protease activities, but a reduction from the early to late passage was also observed. The reduction in the level of protease activity was also observed as a gradual process during on-going culture of lines derived from the Hissar and Ode stocks. In the Ode line we demonstrated a parallel decrease in the production of microschizonts upon temperature shift in vitro.

Keywords

Theileria annulatametalloproteasesvaccineattenuationmerogony
Type
Research Article
Information
Parasitology , Volume 105 , Issue 3 , December 1992 , pp. 417 - 423
Copyright
Copyright © Cambridge University Press 1992

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References

REFERENCES

Anderson, C. W., Baum, P. R. & Gesteland, R. F. (1973). Processing of adenovirus-2 induced proteins. Journal of Virology 12, 241–52.CrossRefGoogle ScholarPubMed
Barrett, A. J. & Rawlings, N. D. (1991). Proteinases. Biochemical Society Transactions 19, 707–15.CrossRefGoogle Scholar
Benyon, R. J. & Bond, J. S. (1989). Proteolytic Enzymes: a Practical Approach. Oxford: IRL Press.Google Scholar
Bond, J. S. & Benyon, R. J. (1985). Mammalian metalloendopeptidases. International Journal of Biochemistry 17, 565–74.CrossRefGoogle ScholarPubMed
Bouvier, J., Schneider, P., Etges, R. & Bordier, C. (1990). Peptide substrate specificity of the membranebound metalloprotease of Leishmania. Biochemistry 29, 10113–19.CrossRefGoogle ScholarPubMed
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–54.CrossRefGoogle ScholarPubMed
Braun-Breton, C., Rosenberry, T. L. & Pereira Da Silva, L. (1988). Induction of the proteolytic activity of a membrane protein in Plasmodium falciparum by phosphatidyl inositol-specific phospholipase C. Nature, London 332, 457–9.CrossRefGoogle ScholarPubMed
Brown, C. G. D. (1987). Theileriidae. In In Vitro Methods for Parasite Cultivation (ed. Taylor, A. E. R. & Baker, J. R.), pp. 230–53. London: Academic Press.Google Scholar
Commins, M. A., Goodger, B. V. & Wright, I. G. (1985). Proteinases in the lysate of bovine erythrocytes infected with Babesia bovis: initial vaccination studies. International Journal for Parasitology 15, 491–5.CrossRefGoogle ScholarPubMed
Gill, B. S., Bhattacharyulu, Y., Kaur, D. & Singh, A. (1976). Vaccination against bovine topical theileriasis (Theileria annulata). Nature, London 264, 355–6.CrossRefGoogle Scholar
Glascodine, J., Tetley, L., Tait, A., Brown, D. & Shiels, B. (1990). Developmental expression of a Theileria annulata merozoite surface antigen. Molecular and Biochemical Parasitology 40, 105–12.CrossRefGoogle ScholarPubMed
Hall, F. R. (1988). Antigens and immunity in Theileria annulata. Parasitology Today 4, 257–61.CrossRefGoogle ScholarPubMed
Hashemi-Fesharki, R. (1988). Control of Theileria annulata in Iran. Parasitology Today 4, 36–40.CrossRefGoogle ScholarPubMed
Hotez, P., Haggerty, J., Hawdon, J., Milstone, L., Gamble, H. R., Schad, G. & Richards, F. C. S. (1990). Metalloproteases of infective Ancylostoma hookworm larvae and their possible functions in tissue invasion and ecdysis. Infection and Immunity 58, 3883–92.CrossRefGoogle ScholarPubMed
Hulliger, L., Brown, C. G. D. & Wilde, J. K. H. (1966). Transition of developmental stages of Theileria parva in vitro at high temperature. Nature, London 261, 311–12.Google Scholar
Keene, W. E., Hidalgo, M. E., Orozco, E. & Mckerrow, J. H. (1990). Entamoeba histolytica: correlation of the cytopathic effect of virulent trophozoites with the secretion of a cysteine proteinase. Experimental Parasitology 71, 199–206.CrossRefGoogle ScholarPubMed
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–5.CrossRefGoogle ScholarPubMed
Lockwood, B. C., North, M. J., Scott, K. I., Bremner, A. F. & Coombs, G. H. (1987). The use of a highly sensitive electrophoretic method to compare the proteinases of trichomonads. Molecular and Biochemical Parasitology 24, 89–95.CrossRefGoogle ScholarPubMed
Marco, M. & Nieto, A. (1991). Metalloproteinases in the larvae of Echinococcus granulosus. International Journal for Parasitology 21, 743–6.CrossRefGoogle ScholarPubMed
Melrose, T. R. & Brown, C. G. D. (1980). Isoenzyme variation in piroplasms isolated from bovine blood infected with Theileria annulata and Theileria parva. Research in Veterinary Science 27, 379–81.CrossRefGoogle Scholar
Morzaria, S. P., Roeder, P. L., Roberts, D. H., Chasey, D. & Drew, T. W. (1982). Characteristics of a continuous suspension cell line derived from a calf with sporadic bovine leukosis. In Fifth International Symposium on Bovine Leukosis (ed. Straub, O. C.), pp. 519–28. Luxembourg: Commission of the European Communities.Google Scholar
Nene, V., Gobright, E., Musoke, A. J. & Lonsdale-Eccles, J. D. (1990). A single exon codes for the enzyme domain of a protozoan cysteine protease. Journal of Biological Chemistry 265, 18047–50.CrossRefGoogle Scholar
Ozkoc, U. & Pipano, E. (1981). Trials with cell culture vaccine against theileriosis in Turkey. In Advances in the Control of Theileriosis (ed. Irvin, A. D., Cummingham, M. P. & Young, A. S.), pp. 256–7. The Hague: Martinus Nijhoff.CrossRefGoogle Scholar
Pipano, E. (1977). Basic principles of Theileria annulata control. In Theileriosis (ed. Henson, J. B. & Campbell, M.), pp. 55–65. Ottawa: International Development Research Center.Google Scholar
Pipano, E. (1981). Schizonts and tick stages in immunisation against Theileria annulata infection. In Advances in the Control of Theileriosis (ed. Irvin, A. D., Cunningham, M. P. & Young, A. S.), pp. 242–252. The Hague: Martinus Nijhoff.CrossRefGoogle Scholar
Pipano, E. (1989). Bovine theileriosis in Israel. Revue Scientifique et Technique, Office International des Epizooties 8, 79–87.CrossRefGoogle ScholarPubMed
Schein, E., Buscher, G. & Friedhoff, K. T. (1975). Lichtmikroskopie Untersuchungen über die Entwicklung von Theileria annulata (Dschunkowsky and Luhs) in Hyalomma anatolicum excavatum (Koch, 1844). Zeitschrift für Parasitenkunde 48, 123–36.CrossRefGoogle Scholar
Shiels, B. R., Mcdougall, C., Tait, A. & Brown, C. G. D. (1986). Identification of infection-associated antigens in Theileria annulata transformed cells. Parasite Immunology 8, 69–77.CrossRefGoogle ScholarPubMed
Shukla, P. C. & Sharma, R. D. (1991). Immunization of bovine calves with cell culture vaccine against Theileria annulata. Acta Veterinaria 60, 79–86.CrossRefGoogle Scholar
Singh, D. K. (1990). Methods currently used for the control of Theileria annulata: their validity and proposals for future control strategies. Parassitologia 32, 33–40.Google ScholarPubMed
Theilen, G. H., Rush, J. D., Nelson-Rees, W. A., Dungworth, D. L., Munn, R. J. & Switzer, J. W. (1968). Bovine leukemia: establishment and morphological characterisation of continuous cell suspension culture, BL-1. Journal of the National Cancer Institute 40, 737–49.Google ScholarPubMed
Woessner, J. F. (1991). Matrix metalloproteinases and their inhibitors in connective tissue remodelling. FASEB Journal 5, 2145–54.CrossRefGoogle Scholar
Wright, I. G., Goodger, B. V. & Mahoney, D. F. (1981). Virulent and avirulent strains of Babesia bovis: the relationship between parasite protease content and pathophysiological effect of the strain. Journal of Protozoology 28, 118–20.CrossRefGoogle ScholarPubMed