In order to effectively control and eliminate rinderpest, a method is required to allow serological differentiation between animals that have been vaccinated and those which have recovered from natural infection. One way of doing this would be to engineer the normal vaccine to produce a genetically marked rinderpest virus (RPV) vaccine. We constructed two modified cDNA clones of the RPV RBOK vaccine strain with the coding sequence of the green fluorescent protein (GFP) gene inserted as a potential genetic marker. RPVINS-GFP virus was designed to produce independent and high level expression of GFP inside infected cells, whilst the GFP expressed by RPVSIG-GFP virus was designed to be efficiently secreted. Infectious recombinant virus was rescued in cell culture from both constructs. The effectiveness of these viruses in stimulating protective immunity and antibody responses to the marker protein was tested by vaccination of cattle and goats. All of the vaccinated animals were completely protected when challenged with virulent virus: RPV in cattle or peste-des-petits ruminants virus in the goats. ELISA showed that all of the animals produced good levels of anti-RPV antibodies. Three of the four cattle and the two goats vaccinated with RPVSIG-GFP produced detectable levels of anti-GFP antibodies. In contrast, no anti-GFP antibodies were produced in the four cattle and two goats vaccinated with RPVINS-GFP. Therefore, secretion of the GFP marker protein was absolutely required to elicit an effective humoral antibody response to the marker protein.


Article metrics loading...

Loading full text...

Full text loading...



  1. Anderson, J. & McKay, J. A. (1994). The detection of antibodies against peste des petits ruminants virus in cattle, sheep and goats and the possible implications to rinderpest control programmes.Epidemiology and Infection 112, 225-234.[CrossRef] [Google Scholar]
  2. Anderson, J., Barrett, T. & Scott, G. R. (1996).FAO Animal Health Manual. Manual on the Diagnosis of Rinderpest, 2nd edn. Rome: Food and Agriculture Organization of the United Nations.
  3. Anon. (1998). Recommended standards for epidemiological surveillance system for rinderpest. Revue Scientifique et Technical de l’Office International des Epizooties 17, 825–828. [Google Scholar]
  4. Baron, M. D. & Barrett, T. (1997). Rescue of rinderpest virus from cloned cDNA.Journal of Virology 71, 1265-1271. [Google Scholar]
  5. Baron, M. D., Ebel, T. & Suomalainen, M. (1992). Intracellular transport of rubella virus structural proteins expressed from cloned cDNA.Journal of General Virology 73, 1073-1086.[CrossRef] [Google Scholar]
  6. Baron, M. D., Foster-Cuevas, M., Baron, J. & Barrett, T. (1999). Expression of epitopes of a heterologous virus using a recombinant rinderpest virus.Journal of General Virology 80, 2031-2039. [Google Scholar]
  7. Boyle, J. S., Brady, J. L. & Lew, A. M. (1998). Enhanced responses to a DNA vaccine encoding a fusion antigen that is directed to sites of immune induction.Nature 392, 408-411.[CrossRef] [Google Scholar]
  8. Calain, P. & Roux, L. (1993). The rule of six, a basic feature for efficient replication of Sendai virus defective interfering RNA.Journal of Virology 67, 4822-4830. [Google Scholar]
  9. Diallo, A., Barrett, T., Lefevre, P. C. & Taylor, W. P. (1987). Comparison of proteins induced in cells infected with rinderpest and peste des petits ruminants viruses.Journal of General Virology 68, 2033-2038.[CrossRef] [Google Scholar]
  10. Forsyth, M. & Barrett, T. (1995). Evaluation of polymerase chain reaction for the detection and characterization of rinderpest and peste des petits ruminants viruses for epidemiological studies.Virus Research 39, 151-163.[CrossRef] [Google Scholar]
  11. Hasan, M. K., Kato, A., Shioda, T., Sakai, Y., Yu, D. & Nagai, Y. (1997). Creation of an infectious recombinant Sendai virus expressing the firefly luciferase gene from the 3′ proximal first locus.Journal of General Virology 78, 2813-2820. [Google Scholar]
  12. He, B., Paterson, R. G., Ward, C. D. & Lamb, R. A. (1997). Recovery of infectious SV5 from cloned cDNA and expression of a foreign gene.Virology 237, 249-260.[CrossRef] [Google Scholar]
  13. Kobune, F., Sakata, H. & Sugiura, A. (1990). Marmoset lymphoblastoid cells as a sensitive host for isolation of measles virus.Journal of Virology 72, 687-692. [Google Scholar]
  14. Kobune, F., Sakata, H., Sugiyama, M. & Sugiura, A. (1991). B95a, a marmoset lymphoblastoid cell line, as a sensitive host for rinderpest virus.Journal of General Virology 72, 687-692.[CrossRef] [Google Scholar]
  15. Kretzschmar, E., Buonocore, L., Schnell, M. J. & Rose, J. K. (1997). High efficiency incorporation of functional influenza virus glycoproteins into recombinant vesicular stomatitis viruses.Journal of Virology 71, 5982-5989. [Google Scholar]
  16. Mariner, J. C., House, J. A., Mebus, C. A. & Van den Ende, M. C. (1993). The use of thermostable Vero cell-adapted rinderpest vaccine as a heterologous vaccine against peste des petits ruminants.Research in Veterinary Science USA 54, 212-216.[CrossRef] [Google Scholar]
  17. Marks, M. S., Roche, P. A., Van Donselaar, E., Woodruff, L., Peters, P. J. & Bonifacino, J. S. (1995). A lysosomal targetting signal in the cytoplasmic tail of the β chain directs HLA- DM to MHC class II compartments.Journal of Cell Biology 131, 351-369.[CrossRef] [Google Scholar]
  18. Mebatsion, T., Schnell, M. J., Cox, J. H., Finke, S. & Conzelmann, K. (1996). Highly stable expression of a foreign gene from rabies virus vectors.Proceedings of the National Academy of Sciences, USA 93, 7310-7314.[CrossRef] [Google Scholar]
  19. Plowright, W. (1984). The duration of immunity in cattle following inoculation of rinderpest cell culture vaccine.Journal of Hygiene, Cambridge 92, 285-296.[CrossRef] [Google Scholar]
  20. Plowright, W. & Ferris, R. D. (1962). Studies with rinderpest virus in tissue culture. The use of attenuated culture virus as a vaccine in cattle.Research in Veterinary Science 3, 172-182. [Google Scholar]
  21. Roberts, A., Kretzschmar, E., Perkins, A. S., Forman, J., Price, R., Buonocore, L., Kawaoka, Y. & Rose, J. K. (1998). Vaccination with recombinant vesicular stomatitis viruses expressing an influenza virus hemagglutinin provides complete protection from influenza virus challenge.Journal of Virology 72, 4704-4711. [Google Scholar]
  22. Roberts, A., Buonocore, L., Price, R. & Rose, J. K. (1999). Attenuated vesicular stomatitis viruses as vaccine vectors.Journal of Virology 73, 3723-3732. [Google Scholar]
  23. Rweyemamu, M. M. & Cheneau, Y. (1995). Strategy for the global rinderpest eradication programme.Veterinary Microbiology 44, 369-376.[CrossRef] [Google Scholar]
  24. Shaji, D. & Shaila, M. S. (1999). Domains of rinderpest virus phosphoprotein involved in interaction with itself and the nucleocapsid protein.Virology 258, 415-424.[CrossRef] [Google Scholar]
  25. Singh, M. & Billeter, M. (1999). A recombinant measles virus expressing biologically active human interleukin-12.Journal of General Virology 80, 101-106. [Google Scholar]
  26. Singh, M., Cattaneo, R. & Billeter, M. (1999). A recombinant measles virus expressing hepatitis B virus surface antigen induces humoral immune responses in genetically modified mice.Journal of Virology 73, 4823-4828. [Google Scholar]
  27. Taylor, W. P. (1979). Protection of goats against peste des petits ruminants with attenuated rinderpest virus.Research in Veterinary Science 27, 321-324. [Google Scholar]
  28. Taylor, W. P. (1986). Epidemiology and control of rinderpest.Revue Scientifique et Technical de l′Office International des Epizooties 5, 407-410. [Google Scholar]
  29. Taylor, W. P. & Plowright, W. (1965). Studies on the pathogenesis of rinderpest in experimental cattle. III. Proliferation of an attenuated strain in various tissues following subcutaneous inoculation.Journal of Hygiene, Cambridge 63, 263-275.[CrossRef] [Google Scholar]
  30. Taylor, W. P., Bhat, P. N. & Nanda, Y. P. (1995). The principles and practice of rinderpest eradication.Veterinary Microbiology 44, 359-367.[CrossRef] [Google Scholar]
  31. Thompson, S. A., Burrows, S. R., Misko, I. S., Moss, D. J., Coupar, B. E. H. & Khanna, R. (1998). Targeting a polyepitope protein incorporating multiple class II-restricted viral epitopes to the secretory/endocytic pathway facilitates immune recognition by CD4+ cytotoxic T lymphocytes: a novel approach to vaccine design.Journal of Virology 72, 2246-2252. [Google Scholar]
  32. Tsien, R. Y. (1998). The green fluorescent protein.Annual Review of Biochemistry 67, 509-544.[CrossRef] [Google Scholar]
  33. Von Heijne, G. (1983). Patterns of amino acids near signal-sequence cleavage sites.European Journal of Biochemistry 133, 17-21.[CrossRef] [Google Scholar]
  34. Wertz, G. W., Perepelitsa, V. P. & Ball, L. A. (1998). Gene rearrangement attenuates expression and lethality of a nonsegmented negative strand RNA virus.Proceedings of the National Academy of Sciences, USA 95, 3501-3506.[CrossRef] [Google Scholar]
  35. Wohlstein, P., Trautwein, G., Harder, T. C., Leiss, B. & Barrett, T. (1993). Viral antigen distribution in organs of cattle experimentally infected with rinderpest virus.Veterinary Pathology 30, 544-554.[CrossRef] [Google Scholar]

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error