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Expression of p24 gag Protein of Bovine Leukemia Virus in Insect Cells and Its Use in Immunodetection of the Disease

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Abstract

Bovine leukemia is a common retroviral infection of cattle. The disease is characterized by a strong immunological response to several viral proteins, but the antibodies against p24 and gp51 are predominant. In this study, a recombinant baculovirus containing the gag gene p24 was constructed and the protein, used as antigen, analyzed by western blot and an indirect in-house rp24-ELISA test. This allowed detecting the presence of antibodies for bovine leukemia virus in a panel of cattle sera. The authentication of the protein expands its potential use for different medical applications, from improved diagnosis of the disease to source of antigens to be included in a subunit vaccine.

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References

  1. Burny, A., Bruck, C., Cleuter, Y., Couez, D., Deschamps, J., Gregoire, D., et al. (1985). Bovine leukemia virus and enzootic bovine leucosis. Onderstepoort Journal of Veterinary Research, 52(3), 133–144.

    CAS  Google Scholar 

  2. Burridge, M. J., Puhr, D. M., & Henneman, J. M. (1981). Prevalence of bovine leukemia virus infection in Florida. Journal of the American Veterinary Medical Association, 179, 704–707.

    CAS  Google Scholar 

  3. Hopkins, S. G., & DiGiacomo, R. F. (1997). Natural transmission of bovine leukemia virus in dairy and beef cattle. Veterinary Clinics of North America. Food Animal Practice, 13, 107–128.

    CAS  Google Scholar 

  4. Sagata, N., Yasunaga, T., Tsuzuku-Kawamura, J., Ohishi, K., Ogawa, Y., & Ikawa, Y. (1985). Complete nucleotide sequence of the genome of bovine leukemia virus: its evolutionary relationship to other retroviruses. Proceedings of the National Academy of Sciences of the United States of America, 82, 677–681.

    Article  CAS  Google Scholar 

  5. Bex, F., Bruck, C., Mammerickx, M., Portetelle, D., Ghysdael, J., Cleuter, Y., et al. (1979). Humoral antibody response to bovine leukemia virus infection in cattle and sheep. Cancer Research, 39(3), 1118–1123.

    CAS  Google Scholar 

  6. Mammerickx, M., Portetelle, D., Burny, A., & Leunen, J. (1980). Detection by immunodiffusion- and radioimmunoassay-tests of antibodies to bovine leukemia virus antigens in sera of experimentally infected sheep and cattle. Zentralblatt für Veterinärmedizin. Reihe B, 27, 291–303.

    Article  CAS  Google Scholar 

  7. De Boer, G. F., Boerrigter, H. M., Groen, J., & Osterhaus, A. D. (1987). Identification of bovine leukemia virus (BLV) infected cattle by complex-trapping-blocking (CTB) ELISA employing monoclonal antibodies directed against BLV-p24. Zentralblatt für Veterinärmedizin. Reihe B, 34, 717–728.

    Google Scholar 

  8. Kaaden, O. R., Lange, S., Romanowski, W., Marré, H., Pfeilsticker, J., & Roselius, R. (1982). Transient viraemia with bovine leukaemia virus in bulls. Zentralblatt für Veterinärmedizin. Reihe B, 29, 269–274.

    Article  CAS  Google Scholar 

  9. Kittelberger, R., Reichel, M. P., Meynell, R. M., Tham, K. M., & Molloy, J. B. (1999). Detection of antibodies against the core protein p24 of the bovine leukaemia virus in cattle for confirmatory serological testing. Journal of Virological Methods, 77, 109–114.

    Article  CAS  Google Scholar 

  10. Molloy, J. B., Walker, P. J., Baldock, F. C., Rodwell, B. J., & Cowley, J. A. (1990). An enzyme-linked immunosorbent assay for detection of bovine leukaemia virus p24 antibody in cattle. Journal of Virological Methods, 28, 47–57.

    Article  CAS  Google Scholar 

  11. Rohrmann, G. F. (2008). Baculovirus molecular biology [Internet]. Bethesda, MD: National Center for Biotechnology Information (US). Available from: http://www.ncbi.nlm.nih.gov/books/NBK1736/.

  12. Moscardi, F. (1999). Assessment of the application of baculoviruses for control of Lepidoptera. Annual Review of Entomology, 44, 257–289.

    Article  CAS  Google Scholar 

  13. O’Reilly, D. R., Miller, L. K., & Luckow, V. A. (1992). Baculovirus expression vectors: A laboratory manual. New York: Freeman.

    Google Scholar 

  14. Hu, Y. C. (2005). Baculovirus as a highly efficient expression vector in insect and mammalian cells. Acta Pharmacologica Sinica, 26, 405–416.

    Article  CAS  Google Scholar 

  15. Kost, T. A., Condreay, J. P., & Jarvis, D. L. (2005). Baculovirus as versatile vectors for protein expression in insect and mammalian cells. Nature Biotechnology, 23, 567–575.

    Article  CAS  Google Scholar 

  16. Godeau, F., Saucier, C., & Kourilsky, P. (1992). Replication inhibition by nucleoside analogues of a recombinant Autographa californica multicapsid nuclear polyhedrosis virus harboring the herpes thymidine kinase gene driven by the IE-1(0) promoter: A new way to select recombinant baculoviruses. Nucleic Acids Research, 20, 6239–6246.

    Article  CAS  Google Scholar 

  17. Lalumiere, M., & Richardson, C. D. (1995). Production of recombinant baculoviruses using rapid screening vectors that contain the gene for betagalactosidase. Methods in Molecular Biology, 39, 161–177.

    CAS  Google Scholar 

  18. Southern, J. A., Young, D. F., Heaney, F., Baumgärtner, W. K., & Randall, R. E. (1991). Identification of an epitope on the P and V proteins of simian virus 5 that distinguishes between two isolates with different biological characteristics. Journal of General Virology, 72, 1551–1557.

    Article  CAS  Google Scholar 

  19. Gutiérrez, G., Alvarez, I., Fondevila, N., Politzki, R., Lomónaco, M., Rodríguez, S., et al. (2009). Detection of bovine leukemia virus specific antibodies using recombinant p24-ELISA. Veterinary Microbiology, 137, 224–234.

    Article  Google Scholar 

  20. Trono, K. G., Pérez-Filgueira, D. M., Duffy, S., Borca, M. V., & Carrillo, C. (2001). Seroprevalence of bovine leukemia virus in dairy cattle in Argentina: Comparison of sensitivity and specificity of different detection methods. Veterinary Microbiology, 83, 235–248.

    Article  CAS  Google Scholar 

  21. González, E. T., Oliva, G. A., Valera, A., Bonzo, E., Licursi, M., & Etcheverrigaray, M. E. (2001). Leucosis enzoótica bovina: Evaluación de técnicas de diagnóstico (ID, ELISA-I, WB, PCR) en bovinos inoculados experimentalmente. Analecta Veterinaria, 21, 12–20.

    Google Scholar 

  22. Van der Maaten, M., & Miller, J. M. (1990). Bovine leukosis virus. In Z. Dinter & B. Morein (Eds.), Virus infections of ruminants (pp. 419–429). Amsterdam, The Netherlands: Elsevier.

    Google Scholar 

  23. Bhatia, S., Patil, S. S., Sood, R., Dubey, R., Bhatia, A. K., Pattnaik, B., et al. (2008). Prokaryotic expression of a 750 bp capsid region of bovine immunodeficiency virus gag gene and development of a recombinant capsid (p26) protein based immunoassay for seroprevalence studies. Indian Journal of Biotechnology, 7, 34–45.

    Google Scholar 

  24. Costa, M., García, L., Yunus, A. S., Rockemann, D. D., Samal, S. K., & Cristina, J. (2000). Bovine respiratory syncytial virus: First serological evidence in Uruguay. Veterinary Research, 31, 241–246.

    Article  CAS  Google Scholar 

  25. Bicka, L., Kuźmak, J., Kozaczyńska, B., Plucienniczak, A., & Skorupska, A. (2001). Expression of bovine leukemia virus protein p24 in Escherichia coli and its use in the immunoblotting assay. Acta Biochimica Polonica, 48, 227–232.

    CAS  Google Scholar 

  26. Juliarena, M., Gutierrez, S., & Ceriani, C. (2007). Chicken antibodies: A useful tool for antigen capture ELISA to detect bovine leukaemia virus without cross-reaction with other mammalian antibodies. Veterinary Research Communications, 31, 43–51.

    Article  CAS  Google Scholar 

  27. Momtaz, H., Hemmatzadeh, F., & Keyvanfar, H. (2008). Expression of bovine leukemia virus p24 protein in bacterial cell. Pakistan Journal of Biological Sciences, 11, 2433–2437.

    Article  CAS  Google Scholar 

  28. Zajac, V., & Sláviková, K. (1989). Expression of a bovine leukaemia virus envelope fusion protein in E. coli. Folia Biologica, 35, 35–41.

    CAS  Google Scholar 

  29. Dumont, J., Legrain, M., Portetelle, D., Brasseur, R., Burny, A., & Hilger, F. (1989). High yield synthesis of the bovine leukemia virus (BLV) p24 major internal protein in Saccharomyces cerevisiae. Gene, 79, 219–226.

    Article  CAS  Google Scholar 

  30. Morton, C. L., & Potter, P. M. (2000). Comparison of Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris, Spodoptera frugiperda, and COS7 cells for recombinant gene expression: Application to a rabbit liver carboxylesterase. Molecular Biotechnology, 16, 193–202.

    Article  CAS  Google Scholar 

  31. Dube, S., Dolcini, G., Abbott, L., Mehta, S., Dube, D., Gutierrez, S., et al. (2000). The complete genomic sequence of a BLV strain from a Holstein cow from Argentina. Virology, 277, 379–386.

    Article  CAS  Google Scholar 

  32. Amborski, G. F., Lo, J. L., & Seger, C. L. (1989). Serological detection of multiple retroviral infections in cattle: Bovine leukemia virus, bovine syncytial virus and bovine visna virus. Veterinary Microbiology, 20, 247–253.

    Article  CAS  Google Scholar 

  33. Heeney, J., Valli, V., & Montesanti, J. (1988). Alteration in humoral immune response to bovine leukaemia virus antigens in cattle with lymphoma. Journal of General Virology, 69, 659–666.

    Article  CAS  Google Scholar 

  34. Valker, P., Molloy, J., & Rodwell, B. (1987). A protein immunoblot test for detection of bovine leukemia virus p24 antibody in cattle and experimentally infected sheep. Journal of Virological Methods, 15, 201–211.

    Article  Google Scholar 

  35. Bünger, I., Khalaf, H., Cripe, C., & Rimpler, M. (1994). Detection of antibodies against the virus of enzootic bovine leukosis in serum and milk samples using an immunoblot. DTW. Deutsche Tierarztliche Wochenschrift, 101, 402–405.

    Google Scholar 

  36. Grover, Y., & Guillemain, B. (1992). An immunoblotting procedure for detection of antibodies against bovine leukemia virus in cattle. Journal of Veterinary Medicine, 39, 48–52.

    Article  CAS  Google Scholar 

  37. Merza, M., Sundquist, B., Söber, J., & Morein, B. (1991). Immunoaffinity purification of two major proteins of bovine leukemia virus (gp51 and p24) and their use for discrimination between vaccinated and infected animals. Journal of Virological Methods, 33, 345–353.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Faculty of Veterinary Sciences of La Plata University, Argentina. The authors thank Dr Don Jarvis and Christoph Geisler from Wyoming University, USA for their help in the baculovirus expression system.

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Authors and Affiliations

  1. Veterinary Immunology, Faculty of Veterinary Sciences, University of La Plata, UNLP, 60 y 118, 1900, La Plata, Argentina

    Alejandra Larsen, María Soledad Serena & Eduardo Mortola

  2. Virology, Faculty of Veterinary Sciences, University of La Plata, 60 y 118, 1900, La Plata, Argentina

    Ester Teresa Gonzalez, María Soledad Serena & María Gabriela Echeverría

  3. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Av. Rivadavia, 1917, CABA, Argentina

    María Soledad Serena & María Gabriela Echeverría

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  1. Alejandra Larsen
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  2. Ester Teresa Gonzalez
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  3. María Soledad Serena
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  4. María Gabriela Echeverría
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  5. Eduardo Mortola
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Correspondence to Eduardo Mortola.

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Larsen, A., Gonzalez, E.T., Serena, M.S. et al. Expression of p24 gag Protein of Bovine Leukemia Virus in Insect Cells and Its Use in Immunodetection of the Disease. Mol Biotechnol 54, 475–483 (2013). https://doi.org/10.1007/s12033-012-9587-7

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