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Recombinant subunit vaccines against Neospora caninum

Pinitkiatisakul, Sunan (2007). Recombinant subunit vaccines against Neospora caninum. Diss. (sammanfattning/summary) Uppsala : Sveriges lantbruksuniv., Acta Universitatis Agriculturae Sueciae, 1652-6880 ; 2007:40
ISBN 978-91-576-7339-8
[Doctoral thesis]

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The intracellular protozoan parasite Neospora caninum is an important cause of bovine abortion and congenital infection in many countries. Currently there is no effective control method available, and development of a vaccine has been suggested as a possible strategy to prevent the disease neosporosis. In this thesis an experimental recombinant subunit vaccine, consisting of the N. caninum antigen NcSRS2 and immunostimulating complexes (iscoms), was evaluated. The iscom is an adjuvant formulation into which antigens can be incorporated by hydrophobic interactions. Many recombinant proteins are hydrophilic, and special methods are needed for binding them to the iscoms and to achieve optimal adjuvant effects. A novel method to combine recombinant proteins with iscoms utilizing the strong interaction between biotin and streptavidin was evaluated. Recombinant NcSRS2 was expressed in Escherichia coli as two different fusion proteins. One of them was biotinylated in E. coli and was bound to streptavidin-coated iscom matrix (iscom particles without any antigen). The other fusion protein contained recombinant streptavidin and was bound to biotinylated iscom matrix. These two strategies both resulted in efficient binding to the iscom matrix. Thus, this method might offer a convenient and effective alternative for association of other hydrophilic recombinant antigens with iscoms for evaluation in immunisation experiments. Both the immunogenicity and the protective effect of the NcSRS2-iscoms were investigated by immunisation and challenge infection of mice. A real-time PCR assay was developed, and used for quantification of parasite DNA in the brain and blood of infected mice. The NcSRS2-iscoms induced production of antibodies that recognised NcSRS2 of parasite origin, suggesting that the conformation of the recombinant protein was similar to that of the native antigen. Antigen-specific cellular responses were also induced as demonstrated by in vitro proliferation and cytokine production. After challenge infection, the immunised mice had significantly lower levels of N. caninum DNA in their brains and blood than did non-immunised control mice. These mice were also less affected by the disease, as judged by clinical symptoms and changes in body weight. These results, together with results from other studies, indicate that recombinant NcSRS2 might be considered as a potential candidate antigen for a future subunit vaccine against N. caninum infection in cattle.

Authors/Creators:Pinitkiatisakul, Sunan
Title:Recombinant subunit vaccines against Neospora caninum
Series Name/Journal:Acta Universitatis Agriculturae Sueciae
Year of publishing :April 2007
Number of Pages:53
ALLI. Wikman, M., Friedman, M., Pinitkiatisakul, S., Hemphill, A., Lövgren- Bengtsson, K., Lundén, A. and Ståhl, S. 2005. Applying biotin-streptavidin binding for iscom (immunostimulating complex) association of recombinant immunogens. Biotechnology and Applied Biochemistry 41, 163-74. II. Pinitkiatisakul, S., Mattsson, J.G., Wikman, M., Friedman, M., Lövgren Bengtsson, K., Ståhl, S. & Lundén, A. 2005. Immunisation of mice against neosporosis with recombinant NcSRS2 iscoms. Veterinary Parasitology 129, 25-34. III. Pinitkiatisakul, S., Friedman, M., Wikman, M., Mattsson, J.G., Lövgren- Bengtsson, K., Ståhl, S. & Lundén, A. Immunogenicity and protective effect against murine cerebral neosporosis of recombinant NcSRS2 in different iscom formulations. Vaccine, in press, doi:10.1016/j.vaccine.2007.01.074. IV. Pinitkiatisakul, S., Mattsson, J.G. & Lundén, A. Quantitative analysis of parasite DNA in the blood of immunised and naïve mice after infection with Neospora caninum. Manuscript.
Place of Publication:Uppsala
ISBN for printed version:978-91-576-7339-8
Publication Type:Doctoral thesis
Full Text Status:Public
Agrovoc terms:neospora caninum, parasitoses, immunization, immunostimulants, adjuvants, vaccines, pcr, evaluation, cattle
Keywords:Neospora caninum, NcSRS2, iscom, subunit vaccine, recombinant immunogen, parasitaemia, real-time PCR
Permanent URL:
ID Code:1376
Department:(VH) > Dept. of Biomedical Sciences and Veterinary Public Health
Deposited By: Sunan Pinitkiatisakul
Deposited On:10 Apr 2007 00:00
Metadata Last Modified:02 Dec 2014 10:11

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