Hostname: page-component-586b7cd67f-rcrh6 Total loading time: 0 Render date: 2024-11-28T23:05:45.709Z Has data issue: false hasContentIssue false
  • English
  • Français

Comparative efficacy of immunization with inactivated whole tachyzoites versus a tachyzoite-bradyzoite mixture against neosporosis in mice

Published online by Cambridge University Press:  03 August 2011

S. ROJO-MONTEJO ,
E. COLLANTES-FERNÁNDEZ ,
S. RODRÍGUEZ-MARCOS ,
F. PÉREZ-ZABALLOS ,
I. LÓPEZ-PÉREZ ,
A. PRENAFETA  and
L. M. ORTEGA-MORA
S. ROJO-MONTEJO
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
E. COLLANTES-FERNÁNDEZ
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
S. RODRÍGUEZ-MARCOS
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
F. PÉREZ-ZABALLOS
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
I. LÓPEZ-PÉREZ
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
A. PRENAFETA
Affiliation:
HIPRA, Avda de La Selva 135, Amer 17170, Girona, Spain
L. M. ORTEGA-MORA*
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
*
*Corresponding author: SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040-Madrid, Spain. Tel: +34 913944069. Fax: +34 913944098. E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Summary

The worldwide economic impact of Neospora caninum infection has caused the development of effective vaccines to become one of the main goals in the field of neosporosis research. In this study, the protection conferred by antigens from inactivated whole tachyzoites (TZ) and a tachyzoite-bradyzoite mixture (TZ-BZ) of N. caninum (Nc-Spain7 isolate) incorporated into a water-in-oil emulsion (W/O) and aluminium hydroxide-ginseng extract (Al/G) was evaluated in mouse models of congenital and cerebral N. caninum infection. Immunization with TZ-BZ induced congenital and cerebral neosporosis exacerbation that was mainly characterized by reduced neonatal median survival time and increased parasite presence in adult mouse brains. The immune response of mice immunized with TZ-BZ was characterized by an increase in IFN-γ expression prior to challenge and an increase in IL-4 expression accompanied with significantly higher levels of antibodies against 2 recombinant bradyzoite-specific proteins (rNcSAG4 and rNcBSR4) after challenge. Immunization with TZ in W/O significantly reduced neonatal mortality, vertical transmission as well as parasite presence in adult mouse brains and induced a strong humoral immune response. The current study demonstrates the critical role of stage-specific antigens and adjuvants on the development of effective inactivated vaccines for the prevention of N. caninum infection.

Keywords

Neospora caninuminactivated vaccinetachyzoitebradyzoitemice
Type
Research Article
Information
Parasitology , Volume 138 , Issue 11 , September 2011 , pp. 1372 - 1383
Copyright
Copyright © Cambridge University Press 2011

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Aguado-Martínez, A., Álvarez-García, G., Fernández-García, A., Risco-Castillo, V., Arnaiz-Seco, I., Rebordosa-Trigueros, X., Navarro-Lozano, V. and Ortega-Mora, L. M. (2008). Usefulness of rNcGRA7- and rNcSAG4-based ELISA tests for distinguishing primo-infection, recrudescence, and chronic bovine neosporosis. Veterinary Parasitology 157, 182–195. doi: 10.1016/j.vetpar.2008.08.002.Google Scholar
Aguado-Martínez, A., Álvarez-García, G., Fernández-García, A., Risco-Castillo, V., Marugán-Hernández, V. and Ortega-Mora, L. M. (2009 a). Failure of a vaccine using immunogenic recombinant proteins rNcSAG4 and rNcGRA7 against neosporosis in mice. Vaccine 27, 7331–7338. doi: 10.1016/j.vaccine.2009.09.050.CrossRefGoogle ScholarPubMed
Aguado-Martínez, A., Ortega-Mora, L. M., Álvarez-García, G., Rodríguez-Marco, S., Risco-Castillo, V., Marugán-Hernández, V. and Fernández-García, A. (2009 b). Stage-specific expression of Nc SAG4 as a marker of chronic Neospora caninum infection in a mouse model. Parasitology 136, 757–764. doi: 10.1017/S0031182009006076.CrossRefGoogle ScholarPubMed
Álvarez-García, G., Pitarch, A., Zaballos, A., Fernández-García, A., Gil, C., Gómez-Bautista, M., Aguado-Martínez, A. and Ortega-Mora, L. M. (2007). The NcGRA7 gene encodes the immunodominant 17 kDa antigen of Neospora caninum. Parasitology 134, 41–50. doi: 10.1017/S0031182006001284.Google Scholar
Andrianarivo, A. G., Rowe, J. D., Barr, B. C., Anderson, M. L., Packham, A. E., Sverlow, K. W., Choromanski, L., Loui, C., Grace, A. and Conrad, P. A. (2000). A POLYGEN-adjuvanted killed Neospora caninum tachyzoite preparation failed to prevent foetal infection in pregnant cattle following i.v./i.m. experimental tachyzoite challenge. International Journal for Parasitology 30, 985–990.CrossRefGoogle ScholarPubMed
Augustine, P. C., Jenkins, M. C. and Dubey, J. P. (1999). Effect of polyclonal antisera developed against dense granule-associated Neospora caninum proteins on cell invasion and development in vitro by N. caninum tachyzoites. Parasitology 119, 441–445.Google Scholar
Barber, J. S., Holmdahl, O. J., Owen, M. R., Guy, F., Uggla, A. and Trees, A. J. (1995). Characterization of the first European isolate of Neospora caninum (Dubey, Carpenter, Speer, Topper and Uggla). Parasitology 111, 563–568.CrossRefGoogle ScholarPubMed
Bland, J. M. and Altman, D. G. (2004). The logrank test. BMJ (Clinical Research Ed.) 328, 1073. doi: 10.1136/bmj.328.7447.1073.CrossRefGoogle ScholarPubMed
Bland, J. M. and Altman, D. G. (1998). Survival probabilities (the Kaplan-Meier method). British Medical Journal 317, 1572.CrossRefGoogle ScholarPubMed
Buxton, D., Maley, S. W., Wright, S., Thomson, K. M., Rae, A. G. and Innes, E. A. (1998). The pathogenesis of experimental neosporosis in pregnant sheep. Journal of Comparative Pathology 118, 267–279.Google Scholar
Buxton, D., McAllister, M. M. and Dubey, J. P. (2002). The comparative pathogenesis of neosporosis. Trends in Parasitology 18, 546–552.CrossRefGoogle ScholarPubMed
Cannas, A., Naguleswaran, A., Muller, N., Eperon, S., Gottstein, B. and Hemphill, A. (2003). Vaccination of mice against experimental Neospora caninum infection using NcSAG1- and NcSRS2-based recombinant antigens and DNA vaccines. Parasitology 126, 303–312.Google Scholar
Cho, J. H., Chung, W. S., Song, K. J., Na, B. K., Kang, S. W., Song, C. Y. and Kim, T. S. (2005). Protective efficacy of vaccination with Neospora caninum multiple recombinant antigens against experimental Neospora caninum infection. The Korean Journal of Parasitology 43, 19–25.Google Scholar
Collantes-Fernández, E., López-Pérez, I., Álvarez-García, G. and Ortega-Mora, L. M. (2006). Temporal distribution and parasite load kinetics in blood and tissues during Neospora caninum infection in mice. Infection and Immunity 74, 2491–2494. doi: 10.1128/IAI.74.4.2491-2494. 2006.CrossRefGoogle ScholarPubMed
Di Cristina, M., Del Porto, P., Buffolano, W., Beghetto, E., Spadoni, A., Guglietta, S., Piccolella, E., Felici, F. and Gargano, N. (2004). The Toxoplasma gondii bradyzoite antigens BAG1 and MAG1 induce early humoral and cell-mediated immune responses upon human infection. Microbes and Infection/Institut Pasteur 6, 164–171. doi: 10.1016/j.micinf. 2003.11.009.Google Scholar
Dubey, J. P., Schares, G. and Ortega-Mora, L. M. (2007). Epidemiology and control of neosporosis and Neospora caninum. Clinical Microbiology Reviews 20, 323–367. doi: 10.1128/CMR.00031-06.Google Scholar
Feng, X., Zhang, N. and Tuo, W. (2010). Neospora caninum tachyzoite- and antigen-stimulated cytokine production by bone marrow-derived dendritic cells and spleen cells of naive BALB/c mice. The Journal of Parasitology 96, 717–723. doi: 10.1645/GE-1900.1.Google Scholar
Fernández-García, A., Risco-Castillo, V., Zaballos, A., Álvarez-García, G. and Ortega-Mora, L. M. (2006). Identification and molecular cloning of the Neospora caninum SAG4 gene specifically expressed at bradyzoite stage. Molecular and Biochemical Parasitology 146, 89–97. doi: 10.1016/j.molbiopara.2005.08.019.Google Scholar
Fuchs, N., Sonda, S., Gottstein, B. and Hemphill, A. (1998). Differential expression of cell surface- and dense granule-associated Neospora caninum proteins in tachyzoites and bradyzoites. The Journal of Parasitology 84, 753–758.CrossRefGoogle ScholarPubMed
Hemphill, A., Gottstein, B. and Kaufmann, H. (1996). Adhesion and invasion of bovine endothelial cells by Neospora caninum. Parasitology 112 (Pt 2), 183–197.Google Scholar
Hondowicz, B. and Scott, P. (2002). Influence of parasite load on the ability of type 1 T cells to control Leishmania major infection. Infection and Iimmunity 70, 498–503.CrossRefGoogle ScholarPubMed
Innes, E. A., Andrianarivo, A. G., Bjorkman, C., Williams, D. J. and Conrad, P. A. (2002). Immune responses to Neospora caninum and prospects for vaccination. Trends in Parasitology 18, 497–504.Google Scholar
Innes, E. A. and Vermeulen, A. N. (2006). Vaccination as a control strategy against the coccidial parasites Eimeria, Toxoplasma and Neospora. Parasitology 133 (Suppl), S145–S168. doi: 10.1017/S0031182006001855.CrossRefGoogle ScholarPubMed
Jenkins, M. C., Wouda, W. and Dubey, J. P. (1997). Serological response over time to recombinant Neospora caninum antigens in cattle after a neosporosis-induced abortion. Clinical and Diagnostic Laboratory Immunology 4, 270–274.Google Scholar
Khan, I. A., Schwartzman, J. D., Fonseka, S. and Kasper, L. H. (1997). Neospora caninum: role for immune cytokines in host immunity. Experimental Parasitology 85, 24–34. doi: 10.1006/expr.1996.4110.CrossRefGoogle ScholarPubMed
Kim, S. K. and Boothroyd, J. C. (2005). Stage-specific expression of surface antigens by Toxoplasma gondii as a mechanism to facilitate parasite persistence. Journal of Immunology 174, 8038–8048.Google Scholar
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. doi: 10.1016/j.ijpara. 2006.11.002.Google Scholar
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. The Journal of Parasitology 85, 1072–1075.CrossRefGoogle ScholarPubMed
Livak, K. J. and Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods (San Diego, Calif.) 25, 402–408. doi: 10.1006/meth.2001.1262.CrossRefGoogle ScholarPubMed
Long, M. T., Baszler, T. V. and Mathison, B. A. (1998). Comparison of intracerebral parasite load, lesion development, and systemic cytokines in mouse strains infected with Neospora caninum. The Journal of Parasitology 84, 316–320.Google Scholar
López-Pérez, I. C., Collantes-Fernández, E., Aguado-Martínez, A., Rodríguez-Bertos, A. and Ortega-Mora, L. M. (2008). Influence of Neospora caninum infection in BALB/c mice during pregnancy in post-natal development. Veterinary Parasitology 155, 175–183. doi: 10.1016/j.vetpar. 2008.05.018.Google Scholar
López-Pérez, I. C., Collantes-Fernández, E., Rojo-Montejo, S., Navarro-Lozano, V., Risco-Castillo, V., Pérez-Pérez, V., Pereira-Bueno, J. and Ortega-Mora, L. M. (2010). Effects of Neospora caninum infection at mid-gestation on placenta in a pregnant mouse model. The Journal of Parasitology 96, 1017–1020. doi: 10.1645/GE-2347.1.Google Scholar
López-Pérez, I. C., Risco-Castillo, V., Collantes-Fernández, E. and Ortega-Mora, L. M. (2006). Comparative effect of Neospora caninum infection in BALB/c mice at three different gestation periods. The Journal of Parasitology 92, 1286–1291.Google Scholar
Lunden, A., Wright, S., Allen, J. E. and Buxton, D. (2002). Immunisation of mice against neosporosis. International Journal for Parasitology 32, 867–876.CrossRefGoogle ScholarPubMed
Marugán-Hernández, V., Álvarez-García, G., Risco-Castillo, V., Regidor-Cerrillo, J. and Ortega-Mora, L. M. (2010). Identification of Neospora caninum proteins regulated during the differentiation process from tachyzoite to bradyzoite stage by DIGE. Proteomics 10, 1740–1750. doi: 10.1002/pmic.200900664.Google Scholar
McAllister, M. M., Parmley, S. F., Weiss, L. M., Welch, V. J. and McGuire, A. M. (1996). An immunohistochemical method for detecting bradyzoite antigen (BAG5) in Toxoplasma gondii-infected tissues cross-reacts with a Neospora caninum bradyzoite antigen. The Journal of Parasitology 82, 354–355.Google Scholar
Nishikawa, Y., Tragoolpua, K., Inoue, N., Makala, L., Nagasawa, H., Otsuka, H. and Mikami, T. (2001 a). In the absence of endogenous gamma interferon, mice acutely infected with Neospora caninum succumb to a lethal immune response characterized by inactivation of peritoneal macrophages. Clinical and Diagnostic Laboratory Immunology 8, 811–816. doi: 10.1128/CDLI.8.4.811-817.2001.Google Scholar
Nishikawa, Y., Xuan, X., Nagasawa, H., Igarashi, I., Fujisaki, K., Otsuka, H. and Mikami, T. (2001 b). Prevention of vertical transmission of Neospora caninum in BALB/c mice by recombinant vaccinia virus carrying NcSRS2 gene. Vaccine 19, 1710–1716.Google Scholar
Pérez-Zaballos, F. J., Ortega-Mora, L. M., Álvarez-García, G., Collantes-Fernández, E., Navarro-Lozano, V., García-Villada, L. and Costas, E. (2005). Adaptation of Neospora caninum isolates to cell-culture changes: an argument in favor of its clonal population structure. The Journal of Parasitology 91, 507–510.Google Scholar
Regidor-Cerrillo, J., Gómez-Bautista, M., Del Pozo, I., Jiménez-Ruiz, E., Aduriz, G. and Ortega-Mora, L. M. (2010). Influence of Neospora caninum intra-specific variability in the outcome of infection in a pregnant BALB/c mouse model. Veterinary Research 41, 52–64. doi: 10.1051/vetres/2010024.Google Scholar
Regidor-Cerrillo, J., Gómez-Bautista, M., Pereira-Bueno, J., Aduriz, G., Navarro-Lozano, V., Risco-Castillo, V., Fernández-García, A., Pedraza-Díaz, S. and Ortega-Mora, L. M. (2008). Isolation and genetic characterization of Neospora caninum from asymptomatic calves in Spain. Parasitology 135, 1651–1659. doi: 10.1017/S003118200800509X.CrossRefGoogle ScholarPubMed
Reichel, M. P. and Ellis, J. T. (2009). Neospora caninum–how close are we to development of an efficacious vaccine that prevents abortion in cattle? International Journal for Parasitology 39, 1173–1187. doi: 10.1016/ j.ijpara.2009.05.007.Google Scholar
Reichel, M. P. and Ellis, J. T. (2006). If control of Neospora caninum infection is technically feasible does it make economic sense? Veterinary Parasitology 142, 23–34. doi: 10.1016/j.vetpar.2006.06.027.Google Scholar
Ribeiro, D. P., Freitas, M. M., Cardoso, M. R., Pajuaba, A. C., Silva, N. M., Mineo, T. W., Silva, J. S., Mineo, J. R. and Silva, D. A. (2009). CpG-ODN combined with Neospora caninum lysate, but not with excreted-secreted antigen, enhances protection against infection in mice. Vaccine 27, 2570–2579. doi: 10.1016/j.vaccine.2009.02.028.Google Scholar
Risco-Castillo, V., Fernández-García, A. and Ortega-Mora, L. M. (2004). Comparative analysis of stress agents in a simplified in vitro system of Neospora caninum bradyzoite production. The Journal of Parasitology 90, 466–470.CrossRefGoogle Scholar
Risco-Castillo, V., Fernández-García, A., Zaballos, A., Aguado-Martínez, A., Hemphill, A., Rodríguez-Bertos, A., Álvarez-García, G. and Ortega-Mora, L. M. (2007). Molecular characterisation of BSR4, a novel bradyzoite-specific gene from Neospora caninum. International Journal for Parasitology 37, 887–896. doi: 10.1016/j.ijpara. 2007.02.003.Google Scholar
Rojo-Montejo, S., Collantes-Fernández, E., Regidor-Cerrillo, J., Rodríguez-Bertos, A., Prenafeta, A., Gómez-Bautista, M. and Ortega-Mora, L. M. (2011). Influence of adjuvant and antigen dose on protection induced by an inactivated whole vaccine against Neospora caninum infection in mice. Veterinary Parasitology 175, 220–229. doi: 10.1016/j.vetpar.2010.10.028.Google Scholar
Srinivasan, S., Mueller, J., Suana, A. and Hemphill, A. (2007). Vaccination with microneme protein NcMIC4 increases mortality in mice inoculated with Neospora caninum. The Journal of Parasitology 93, 1046–1055.Google Scholar
Strohbusch, M., Muller, N., Hemphill, A., Margos, M., Grandgirard, D., Leib, S., Greif, G. and Gottstein, B. (2009). Neospora caninum and bone marrow-derived dendritic cells: parasite survival, proliferation, and induction of cytokine expression. Parasite Immunology 31, 366–372. doi: 10.1111/j.1365-3024.2009.01112.x.Google Scholar
Varona, R., Cadenas, V., Gómez, L., Martínez, A. C. and Márquez, G. (2005). CCR6 regulates CD4+ T-cell-mediated acute graft-versus-host disease responses. Blood 106, 18–26. doi: 10.1182/blood-2004-08-2996.Google Scholar
Whitten, M. K. (1957). Effect of exteroceptive factors on the oestrous cycle of mice. Nature, London 180, 1436.Google Scholar
Yamane, I., Kitani, H., Kokuho, T., Shibahara, T., Haritani, M., Hamaoka, T., Shimizu, S., Koiwai, M., Shimura, K. and Yokomizo, Y. (2000). The inhibitory effect of interferon gamma and tumor necrosis factor alpha on intracellular multiplication of Neospora caninum in primary bovine brain cells. The Journal of Veterinary Medical Science/the Japanese Society of Veterinary Science 62, 347–351.Google Scholar