Skip to main content Accessibility help
×
Home

A rapid and sensitive intracellular flow cytometric assay to identify Theileria parva infection within target cells

  • M. S. ROCCHI (a1), K. T. BALLINGALL (a1), D. NGUGI (a2), N. D. MacHUGH (a2) and D. J. McKEEVER (a3)...

Summary

Theileria parva is an intracellular protozoan parasite transmitted by ticks that causes a fatal lymphoproliferative disease of cattle known as East Coast Fever. Vaccination against the disease currently relies on inoculation of the infective sporozoite stage of the parasite and simultaneous treatment with long-acting formulations of oxytetracycline. Sporozoites are maintained as frozen stabilates of triturated infected ticks and the method requires accurate titration of stabilates to determine appropriate dose rates. Titration has traditionally been undertaken in cattle and requires large numbers of animals because of individual variation in susceptibility to infection. An alternative tissue culture-based method is laborious and time consuming. We have developed a flow cytometric method for quantifying the infectivity of sporozoite stabilates in vitro based on the detection of intracellular parasite antigen. The method allows clear identification of parasitized cells with a high degree of sensitivity and specificity. Analysis of infected cells between 48 and 72 h post-infection clearly defines the potential transforming capability of different stabilates.

Copyright

Corresponding author

*Corresponding author: Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik EH26 0PZ, Scotland, UK. Tel: 0044 (0)131 4455111. Fax: 0044 (0)131 4456111. E-mail: Mara.Rocchi@Moredun.ac.uk

References

Hide All
Brown, C. G., Stagg, D. A., Purnell, R. E., Kanhai, G. K. and Payne, R. C. (1973). Letter: Infection and transformation of bovine lymphoid cells in vitro by infective particles of Theileria parva. Nature, London 245, 101103.
Brown, C. G. D. (1987). Theileriidae. In In Vitro Methods for Parasite Cultivation (ed. Taylor, A. E. R. and Baker, J. R.), pp. 230253. Academic Press, New York, USA.
Buscher, G., Morrison, W. I. and Nelson, R. T. (1984). Titration in cattle of infectivity and immunogenicity of autologous cell lines infected with Theileria parva. Veterinary Parasitology 15, 2938.
Cunningham, M. P., Brown, C. G. D., Burridge, M. J. and Purnell, R. E. (1973). Cryopreservation of infective particles of Theileria parva. International Journal for Parasitology 3, 583587.
Fawcett, D. W., Doxsey, S., Stagg, D. A. and Young, A. S. (1982). The entry of sporozoites of Theileria parva into bovine lymphocytes in vitro. Electron microscopic observations. European Journal of Cell Biology 27, 1021.
Goullin, B., Belloc, F., Duiman, P., Masseron, T., Lacombe, F. and Floch, J. J. (1992). Flow cytometric detection of intracellular Leishmania. Biology of the Cell 76, 276.
Malmquist, W. A., Nyindo, M. B. A. and Brown, C. G. (1970). East Coast fever: cultivation in vitro of bovine spleen cell lines infected and transformed by Theileria parva. Tropical Animal Health and Production 2, 139145.
Marcotty, T., Berkvens, D., Besa, R. K., Losson, B., Dolan, T. T., Madder, M., Chaka, G., Van den, B. P. and Brandt, J. (2003). Lyophilisation and resuscitation of sporozoites of Theileria parva: preliminary experiments. Vaccine 22, 213216.
Mbassa, G. K., Kweka, L. E. and Dulla, P. N. (1998). Immunization against East Coast Fever in field cattle with low infectivity Theileria parva stabilate–preliminary assessment. Veterinary Parasitology 77, 4148.
Morrison, W. I., MacHugh, N. D. and Lalor, P. A. (1996). Pathogenicity of Theileria parva is influenced by the host cell type infected by the parasite. Infection and Immunity 64, 557562.
Mukhebi, A. W., Perry, B. D. and Kruska, R. (1992). Estimated economics of theileriosis control in Africa. Preventive Veterinary Medicine 12, 7385.
Pipano, E., Morzaria, S., Spooner, P. and Shkap, V. (2004). Theileriosis. In Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Part 2, section 2.3. OIE, World Organization for Animal Health, Paris, France.
Radley, D. E. (1981). Infection and treatment method of immunization against theileriosis. In Advances in the Control of Theileriosis (ed. Irvin, A. D., Cunningham, M. P. and Young, A. S.), pp. 227237. Martinus Nijhoff Publishers, Boston. USA.
Rocchi, M., Ballingall, K., MacHugh, N. and McKeever, D. (2006). The kinetics of Theileria parva infection and lymphocyte transformation in vitro. International Journal for Parasitology 36, 771778.
Teh, H. S., Harley, E., Philips, R. A. and Miller, R. G. (1977). Quantitative studies on the precursors of cytotoxic lymphocytes. I. Characterization of a clonal assay and determination of the size of clones derived from single precursors. Journal of Immunology 118, 1049–56.
Toye, P., Nyanjui, J., Goddeeris, B. and Musoke, A. J. (1996). Identification of neutralization and diagnostic epitopes on PIM, the polymorphic immunodominant molecule of Theileria parva. Infection and Immunity 64, 18321838.
Walker, A. R., McKellar, S. B., Bell, L. J. and Brown, C. G. D. (1979). Rapid quantitative assessment of Theileria infection in ticks. Tropical Animal Health and Production 11, 2126.
Wilkie, G. M., Kirvar, E. and Brown, C. G. (2002). Validation of an in vitro method to determine infectivity of cryopreserved sporozoites in stabilates of Theileria spp. Veterinary Parasitology 104, 199209.

Keywords

Related content

Powered by UNSILO

A rapid and sensitive intracellular flow cytometric assay to identify Theileria parva infection within target cells

  • M. S. ROCCHI (a1), K. T. BALLINGALL (a1), D. NGUGI (a2), N. D. MacHUGH (a2) and D. J. McKEEVER (a3)...

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed.