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Intraperitoneal and intra-nasal vaccination of mice with three distinct recombinant Neospora caninum antigens results in differential effects with regard to protection against experimental challenge with Neospora caninum tachyzoites

Published online by Cambridge University Press:  16 October 2009

K. DEBACHE ,
C. GUIONAUD ,
F. ALAEDDINE  and
A. HEMPHILL
K. DEBACHE
Affiliation:
Institute of Parasitology, Vetsuisse Faculty, University of Berne, Länggass-Strasse 122, CH-3012Berne, Switzerland
C. GUIONAUD
Affiliation:
Institute of Veterinary Pharmacology & Toxicology, Vetsuisse Faculty, University of Berne, Länggass-Strasse 124, CH-3012Berne, Switzerland
F. ALAEDDINE
Affiliation:
Institute of Veterinary Pharmacology & Toxicology, Vetsuisse Faculty, University of Berne, Länggass-Strasse 124, CH-3012Berne, Switzerland
A. HEMPHILL*
Affiliation:
Institute of Parasitology, Vetsuisse Faculty, University of Berne, Länggass-Strasse 122, CH-3012Berne, Switzerland
*
*Corresponding author: Andrew Hemphill, Institute of Parasitology, Vetsuisse Faculty, University of Berne, Länggass-Strasse 122, CH-3012Berne, Switzerland. Tel. +41 31 6312384. Fax +41 31 6312477. E-mail: hemphill@ipa.unibe.ch
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Summary

Recombinant NcPDI(recNcPDI), NcROP2(recNcROP2), and NcMAG1(recNcMAG1) were expressed in Escherichia coli and purified, and evaluated as potential vaccine candidates by employing the C57Bl/6 mouse cerebral infection model. Intraperitoneal application of these proteins suspended in saponin adjuvants lead to protection against disease in 50% and 70% of mice vaccinated with recNcMAG1 and recNcROP2, respectively, while only 20% of mice vaccinated with recNcPDI remained without clinical signs. In contrast, a 90% protection rate was achieved following intra-nasal vaccination with recNcPDI emulsified in cholera toxin. Only 1 mouse vaccinated intra-nasally with recNcMAG1 survived the challenge infection, and protection achieved with intra-nasally applied recNcROP2 was at 60%. Determination of cerebral parasite burdens by real-time PCR showed that these were significantly reduced only in recNcROP2-vaccinated animals (following intraperitoneal and intra-nasal application) and in recNcPDI-vaccinated mice (intra-nasal application only). Quantification of viable tachyzoites in brain tissue of intra-nasally vaccinated mice showed that immunization with recNcPDI resulted in significantly decreased numbers of live parasites. These data show that, besides the nature of the antigen, the protective effect of vaccination also depends largely on the route of antigen delivery. In the case of recNcPDI, the intra-nasal route provides a platform to generate a highly protective immune response.

Keywords

Recombinant Neospora caninum antigentachyzoitesvaccinationmiceintraperitonealintra-nasal
Type
Research Article
Information
Parasitology , Volume 137 , Issue 2 , February 2010 , pp. 229 - 240
Copyright
Copyright © Cambridge University Press 2009

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References

REFERENCES

Aguado-Martinez, A., Ortege-Mora, L. M., Alvarez-Garcia, G., Rodriguez-Marco, S., Risco-Castillo, V., Marugan-Hernandez, V. and Fernandez-Garcia, A. (2009). Stage-specific expression of NcSAG4 as a marker of chronic Neospora caninum infection in a mouse model. Parasitology 136, 757–764.CrossRefGoogle ScholarPubMed
Alaeddine, F., Keller, N., Leepin, A. and Hemphill, A. (2005). Reduced infection and protection from clinical signs of cerebral neosporosis in C57BL/6 mice vaccinated with recombinant microneme antigen NcMIC1. Journal of Parasitology 91, 657–665.CrossRefGoogle ScholarPubMed
Arakawa, T., Tsuboi, T., Kishimoto, A., Sattabongkot, J., Suwanabun, N., Rungruang, T., Matsumoto, Y., Tsuji, N., Hisaeda, H., Stowers, A., Shimabukuro, I., Sato, Y. and Torii, M. (2003). Serum antibodies induced by intranasal immunization of mice with Plasmodium vivax Pvs25 coadministred with cholera toxin completely block parasite transmission to mosquitoes. Vaccine 21, 3143–3148.CrossRefGoogle Scholar
Arakawa, T., Komesu, A., Otsuki, H., Sattanbongkot, J., Udomsangpetch, R., Matsumoto, Y., Tsuji, N., Wu, Y., Torii, M. and Tsuboi, T. (2005). Nasal immunization with a malaria transmission-blocking vaccine candidate; Pfs25 induces complete protective immunity in mice against field isolates of Plasmodium falciparum. Infection and Immunity 73, 7375–7380.CrossRefGoogle ScholarPubMed
Baszler, T. V., Long, M. T., McElwain, T. F. and Mathison, B. A. (1999). Interferon-gamma and interleukin-12 mediate protection to acute Neospora caninum infection in BALB/c mice. International Journal for Parasitology 29, 1635–1646.CrossRefGoogle ScholarPubMed
Baszler, T. V., McElwain, T. F. and Mathison, B. A. (2000). Immunization of BALB/c mice with killed Neospora caninum tachyzoite antigen induces a type 2 immune response and exacerbates encephalitis and neurological disease. Clinical and Diagnostic Laboroatory Immunology 7, 893–898.CrossRefGoogle ScholarPubMed
Bonenfant, C., Dimier-Poisson, I., Velge-Roussel, F., Buzoni-Gatel, D., Del Giudice, G., Rappuoli, R. and Bout, D. (2001). Intranasal immunization with SAG1 and non-toxic mutant heat-labile enterotoxins protects mice against Toxoplasma gondii. Infection and Immunity 69, 1605–1612.CrossRefGoogle Scholar
Byun, Y., Ohmura, M., Fijihashi, K., Yamamota, S., McGhee, J. R., Udaka, S., Kiyono, H., Takeda, Y., Kohsaka, T. and Yuki, Y. (2001). Nasal immunization with E. coli verotoxin 1 (VT1)-B subunit and a non-toxic mutant of cholera toxin elicits serum neutralizing antibodies. Vaccine 19, 2061–2070.CrossRefGoogle Scholar
Cannas, A., Naguleswaran, A., Müller, N., Gottstein, B., Eperon, S. and Hemphill, A. (2003 a). Vaccination of mice against experimental N. caninum infection using NcSAG1- and NcSRS2-based recombinant antigens and DNA-vaccines. Parasitology 126, 303–312.CrossRefGoogle ScholarPubMed
Cannas, A., Naguleswaran, A., Muller, N., Gottstein, B. and Hemphill, A. (2003 b). Reduced cerebral infection of Neospora caninum-infected mice after vaccination with recombinant microneme protein NcMIC3 and ribi adjuvant. Journal of Parasitology 89, 44–50.CrossRefGoogle ScholarPubMed
Cho, J., Chung, W., Song, K., Na, B., Kang, S., Song, C. and Kim, T. (2005). Protective efficacy of vaccination with Neospora caninum multiple recombinant antigens against experimental Neospora caninum infection. Korean Journal for Parasitology 1, 19–25.CrossRefGoogle Scholar
Debache, K., Alaeddine, F., Guionaud, C., Monney, T., Müller, J., Strohbusch, M., Leib, S. L., Grandgirard, D. and Hemphill, A. (2009). Vaccination with recombinant NcROP2 combined with recombinant NcMIC1 and NcMIC3 reduces cerebral infection and vertical transmission in mice experimentally infected with Neospora caninum tachyzoites. International Journal for Parasitology May 15: Epub ahead of printCrossRefGoogle ScholarPubMed
Debache, K., Guionaud, C., Alaeddine, F., Mevissen, M. and Hemphill, A. (2008). Vaccination of mice with recombinant NcROP2 antigen reduces mortality and cerebral infection in mice infected with Neospora caninum tachyzoites, International Journal for Parasitology 38, 1455–1463.CrossRefGoogle ScholarPubMed
Dimier-Poisson, I., Aline, F., Bout, D. and Mevelec, M. N. (2006). Induction of protective immunity against toxoplasmosis in mice by immunization with Toxoplasma RNA. Vaccine 24, 1705–1709.CrossRefGoogle ScholarPubMed
Dubey, J. P., Hattel, A. L., Lindsay, D. S. and Topper, M. J. (1988). Neonatal Neospora caninum infection in dogs: isolation of the causative agent and experimental transmission. Journal of the American Veterinary Medical Association 193, 1259–1263.Google ScholarPubMed
Dubey, J. P. and Lindsay, D. S. (1996). A review of Neospora caninum and neosporosis. Veterinary Parasitology 67, 1–59.CrossRefGoogle ScholarPubMed
Dubey, J. P., Schares, G. and Ortega, L. M. (2007). Epidemiology and control of neosporosis and Neospora caninum. Clinical Microbiology. Microbiology Reviews 20, 323–367.CrossRefGoogle Scholar
Ellis, J. T., Miller, C., Quinn, H., Ryce, C. and Reichel, M. P. (2008). Evaluation of recombinant proteins of Neospora caninum as vaccine candidates (in a mouse model). Vaccine 26, 5989–5996.CrossRefGoogle ScholarPubMed
Garcia, J. L., Navarro, I. T., Biazzono, L., Freire, R. L., da Silva Guimaraes, J., Cryssafidis, A. L., Bugni, F. M., Leme da Cunha, I. A., Hamada, F. N. and Ferriera Dias, R. C. (2007). Protective activity against oocyst shedding in cats vaccinated with crude rhoptry proteins of the Toxoplasma gondii by the intranasal route. Veterinary Parasitology 145, 197–206.CrossRefGoogle ScholarPubMed
Gondim, L. F., McAllister, M. M., Pitt, W. C. and Zemlicka, D. E. (2004). Coyotes (Canis latrans) are definitive hosts of Neospora caninum. International Journal for Parasitology 34, 159–161.CrossRefGoogle ScholarPubMed
Haldorson, G. J., Mathison, B. A., Wenberg, K., Conrad, P. A., Dubey, J. P., Trees, A. J., Yamane, I. and Baszler, T. V. (2005). Immunization with native surface protein NcSRS2 induces a Th2 immune response and reduces congenital Neospora caninum transmission in mice. International Journal for Parasitology 35, 1407–1415.CrossRefGoogle ScholarPubMed
Hemphill, A., Gottstein, B. and Kaufmann, H. (1996). Adhesion and invasion of bovine endothelial cells by Neospora caninum. Parasitology 112, 183–197.CrossRefGoogle ScholarPubMed
Hemphill, A., Vonlaufen, N. and Naguleswaran, A. (2006). Cellular and immunological basis of the host–parasite relationship during infection with Neospora caninum. Parasitology 133, 261–278.CrossRefGoogle ScholarPubMed
Hirunpetcharat, C., Stanisic, D., Liu, X. Q., Vadolas, J., Strugnell, R. A., Lee, R., Miller, L. H., Kaslow, D. C. and Good, M. F. (1998). Intranasal immunization with yeast-expressed 19 kDa carboxyl-terminal fragment of Plasmodium yoelii merozoite surface protein-1 (yMSP119) induces protective immunity to blood stage malaria infection in mice. Parasite Immunology 20, 413–420.CrossRefGoogle Scholar
Hiroi, T., Iwatani, K., Iijima, H., Kodama, S., Yanagita, M. and Kiyono, H. (1998). Nasal immune system: distinctive Th0 and Th1/Th2 type environments in murine nasal-associated lymphoid tissues and nasal passage, respectively. European Journal of Immunology 28, 3346–3353.3.0.CO;2-P>CrossRefGoogle ScholarPubMed
Igarashi, M., Kano, F., Tamekun, I K., Machado, R. Z., Navarro, I. T., Vidotto, O., Vidotto, M. C. and Garcia, J. L. (2007). Toxoplasma gondii: Evaluation of an intranasal vaccine using recombinant proteins against brain cyst formation in BALB/c mice. Experimental Parasitology 118, 386–392CrossRefGoogle ScholarPubMed
Innes, E. A. (2007). The host–parasite relationship in pregnant cattle infected with Neospora caninum. Parasitology 134, 1903–1910.CrossRefGoogle ScholarPubMed
Innes, E. A. and Mattsson, J. G. (2007). Neospora caninum emerges from the shadow of Toxoplasma gondii. Trends in Parasitology 23, 43–44.CrossRefGoogle ScholarPubMed
Innes, E. A. and Vermeulen, A. N. (2006). Vaccination as a control strategy against the coccidial parasites Eimeria, Toxoplasma and Neospora. Parasitology 133, S145–S168.CrossRefGoogle ScholarPubMed
Khan, I. A., Schwartzman, J. D., Fonseka, S. and Kasper, L. H. (1997). Neospora caninum: role of immune cytokines in host immunity. Experimental Parasitology 85, 23–34.CrossRefGoogle ScholarPubMed
Klevar, S., Kulberg, S., Boysen, P., Storset, A. K., Moldal, T., Bjorkman, C. and Olsen, I. (2007). Natural killer cells act as early responders in an experimental infection with Neospora caninum in calves. International Journal for Parasitology 37, 329–339.CrossRefGoogle Scholar
Liao, M., Ma, L., Bannai, H., Lee, E. G., Xie, Z., Tang, X., Zhang, H., Xuan, X. and Fujisaki, K. (2006). Identification of a protein disulfide isomerase of Neospora caninum in excretory-secretory products and its IgA binding and enzymatic activities. Veterinary Parasitology 139, 47–56.CrossRefGoogle ScholarPubMed
Liddell, S., Jenkins, M. C., Collica, C. M. and Dubey, J. P. (1999). Prevention of vertical transfer of Neospora caninum in BALB/c mice by vaccination. Journal of Parasitology 85, 1072–1075.CrossRefGoogle ScholarPubMed
McAllister, M. M., Dubey, J. P., Lindsay, D. S., Jolley, W. R., Wills, R. A. and McGuire, A. M. (1998). Dogs are definitive hosts of Neospora caninum. International Journal for Parasitology 28, 1473–1478.CrossRefGoogle ScholarPubMed
Meek, B., Back, J. W., Klaren, V. N., Speijer, D. and Peek, R. (2002). Protein disulfide isomerase of Toxoplasma gondii is targeted by mucosal IgA antibodies in humans. FEBS Letters 522, 104–108.CrossRefGoogle ScholarPubMed
Müller, J., Naguleswaran, A., Nüller, N. and Hemphill, A. (2008). Neospora caninum: functional inhibition of protein disulfide isomerase by the broad-spectrum anti-parasitic drug nitazoxanide and other thiazolides. Experminental Parasitology 118, 80–88.CrossRefGoogle ScholarPubMed
Müller, N., Vonlaufen, N., Gianinazzi, C., Leib, S. L. and Hemphill, A. (2002). Application of real time fluorescent PCR for quantitative assessment of Neospora caninum infections in organotypic slice cultures of rat central nervous tissue. Journal of Clinical Microbiology 40, 252–255.CrossRefGoogle Scholar
Naguleswaran, A., Alaeddine, F., Guionaud, C., Vonlaufen, N., Sonda, S., Jenoe, P., Mevissen, M. and Hemphill, A. (2005). Neospora caninum protein disulfide isomerase is involved in tachyzoite-host cell interaction. International Journal for Parasitology 35, 1459–1472.CrossRefGoogle ScholarPubMed
Parmley, S. F., Yang, S., Harth, G., Sibley, L. D., Sucharczuk, A. and Remington, J. S. (1994). Molecular characterization of a 65 kilodalton Toxoplasma gondii antigen expressed abundantly in the matrix of tissue cysts. Molecular and Biochemical Parasitology 66, 283–296.CrossRefGoogle ScholarPubMed
Partidos, C. D., Beignon, A.-S., Semetey, V., Briand, J.-P. and Muller, S. (2001). The bare skin and the nose as non-invasive routes for administering peptide vaccines. Vaccine 19, 2708–2715.CrossRefGoogle Scholar
Porgador, A., Staats, H. F., Faiola, B., Gilboa, E. and Palker, T. J. (1997). Intranasal immunization with CTL epitope peptides from HIV-1 or ovalbumin and the mucosal adjuvant cholera toxin induces peptide-specific CTLs and protection against tumor development in vivo. Journal of Immunology 158, 834–841.CrossRefGoogle ScholarPubMed
Pinto, E. F., Pinheiro, R. O., Rayol, A., Larraga, V. and Rossi-Bergmann, B. (2004). Intranasal vaccination against cutaneous leishmaniasis with a particulated leishmanial antigen or DNA encoding LACK. Infection and Immunity 72, 4521–4527.CrossRefGoogle ScholarPubMed
Ramamoorthy, S., Sanakkayala, N., Vemulapalli, R., Duncan, R. B., Lindsay, D. S., Schurig, G. S., Boyle, S. M., Kasimanickam, R. and Sriranganathan, N. (2007 b). Prevention of lethal experimental infection of C57BL/6 mice by vaccination with Brucella abortus strain RB51 expressing Neospora caninum antigens. International Journal for Parasitology 37, 1521–1529.CrossRefGoogle ScholarPubMed
Ramamoorthy, S., Sanakkayala, N., Vemulapalli, R., Jain, N., Lindsay, D. S., Schurig, G. S., Boyle, S. M. and Sriranganathan, N. (2007 a). Prevention of vertical transmission of Neospora caninum in C57BL/6 mice vaccinated with Brucella abortus strain RB51 expressing N. caninum protective antigens. International Journal for Parasitology 37, 1531–1538.CrossRefGoogle ScholarPubMed
Risco-Castillo, V., Fernández-Garcia, A. and Ortega-Mora, L. M. (2004). Comparative analysis of stress agents in a simplified in vitro system of Neospora caninum bradyzoite production. Journal of Parasitology 90, 466–470.CrossRefGoogle Scholar
Srinivasan, S., Müller, J., Suana, A. and Hemphill, A. (2007). Vaccination with microneme protein NcMIC4 increases mortality in mice inoculated with Neospora caninum. Parasitology 92, 1235–1243.CrossRefGoogle Scholar
Stanley, A. C., Buxton, D., Innes, E. A. and Huntley, J. F. (2004). Intranasal immunisation with Toxoplasma gondii tachyzoite antigen encapsulated into PLG microspheres induces humoral and cell mediated immunity in sheep. Vaccine 22, 3929–3941.CrossRefGoogle ScholarPubMed
Strohbusch, M., Müller, N., Hemphill, A., Greif, G. and Gottstein, B. (2008 a). NcGRA2 as a molecular target to assess the parasiticidal activity of toltrazuril against Neospora caninum. Parasitology 135, 1065–1073.CrossRefGoogle ScholarPubMed
Strohbusch, M., Müller, N., Hemphill, A., Greif, G. and Gottstein, B. (2008 b). NcGRA2-RT-PCR to detect live versus dead parasites in Neospora caninum infected mice. The Open Parasitology Journal 2, 64–68.CrossRefGoogle Scholar
Vonlaufen, N., Guetg, N., Naguleswaran, A., Müller, N., Björkman, C., Schares, G., von Blumröder, D., Ellis, J. and Hemphill, A. (2004). In vitro induction of Neospora caninum bradyzoites in Vero cells reveals differential antigen expression, localization, and host cell recognition of tachyzoites and bradyzoites. Infection and Immunity 72, 576–583.CrossRefGoogle ScholarPubMed
Williams, D. J. L. and Trees, A. J. (2006). Protecting babies: vaccine strategies to prevent foetal infection in Neospora caninum infected cattle. Parasite Immunology 28, 61–67.CrossRefGoogle ScholarPubMed
Williams, D. J., Guy, C. S., Smith, R. F., Ellis, J. T., Björkman, C., Reichel, M. P. and Trees, A. J. (2007). Immunization of cattle with live tachyzoites of Neospora caninum confers protection against fetal death. Infection and Immunity 75, 1343–1348.CrossRefGoogle ScholarPubMed