Vertical transmission from an infected cow to its fetus accounts for the vast majority of new Neospora caninum infections in cattle. A vaccine composed of a chimeric antigen named recNcMIC3-1-R, based on predicted immunogenic domains of the two microneme proteins NcMIC1 and NcMIC3, the rhoptry protein NcROP2, and emulsified in saponin adjuvants, significantly reduced the cerebral infection in non-pregnant BALB/c mice. Protection was associated with a mixed Th1/Th2-type cytokine response. However, the same vaccine formulation elicited a Th2-type immune response in pregnant mice and did not prevent vertical transmission or disease, neither in dams nor in offspring mice. In this study, an alternative vaccine formulation containing recNcMIC3-1-R emulsified in Freund’s incomplete adjuvant, a stimulator of the cellular immunity, was investigated. No protection against vertical transmission and cerebral infection in the pregnant mice and a very limited protective effect in the non-pregnant mice were observed. The vaccine induced a Th1-type immune response characterized by high IgG2a titres and strong IFN-γ expression, which appeared detrimental to pregnancy.
References
[1]
Dubey, J.P.; Schares, G. Neosporosis in animals--the last five years. Vet. Parasitol. 2011, 180, 90–108, doi:10.1016/j.vetpar.2011.05.031.
[2]
Dubey, J.P.; Schares, G.; Ortega-Mora, L.M. Epidemiology and control of neosporosis and Neospora caninum. Clin. Microbiol. Rev. 2007, 20, 323–367, doi:10.1128/CMR.00031-06.
[3]
Hemphill, A.; Gottstein, B. A European perspective on Neospora caninum. Int. J. Parasitol. 2000, 30, 877–924, doi:10.1016/S0020-7519(00)00072-2.
[4]
Reichel, M.P.; Ellis, J.T. If control of Neospora caninum infection is technically feasible does it make economic sense? Vet. Parasitol. 2006, 142, 23–34, doi:10.1016/j.vetpar.2006.06.027.
[5]
Reichel, M.P.; Ellis, J.T. Re-evaluating the economics of neosporosis control. Vet. Parasitol. 2008, 156, 361–362, doi:10.1016/j.vetpar.2008.06.002.
[6]
Davison, H.C.; Otter, A.; Trees, A.J. Estimation of vertical and horizontal transmission parameters of Neospora caninum infections in dairy cattle. Int. J. Parasitol. 1999, 29, 1683–1689, doi:10.1016/S0020-7519(99)00129-0.
[7]
Williams, D.J.; Hartley, C.S.; Bjorkman, C.; Trees, A.J. Endogenous and exogenous transplacental transmission of Neospora caninum - how the route of transmission impacts on epidemiology and control of disease. Parasitology 2009, 136, 1895–1900, doi:10.1017/S0031182009990588.
[8]
Innes, E.A.; Bartley, P.M.; Maley, S.W.; Wright, S.E.; Buxton, D. Comparative host-parasite relationships in ovine toxoplasmosis and bovine neosporosis and strategies for vaccination. Vaccine 2007, 25, 5495–5503, doi:10.1016/j.vaccine.2007.02.044.
[9]
Innes, E.A.; Vermeulen, A.N. Vaccination as a control strategy against the coccidial parasites Eimeria, Toxoplasma and Neospora. Parasitology 2006, 133 (Suppl), S145–S168.
[10]
Reichel, M.P.; Ellis, J.T. Neospora caninum--how close are we to development of an efficacious vaccine that prevents abortion in cattle? Int. J. Parasitol. 2009, 39, 1173–1187, doi:10.1016/j.ijpara.2009.05.007.
Romero, J.J.; Pérez, E.; Frankena, K. Effect of a killed whole Neospora caninum tachyzoite vaccine on the crude abortion rate of Costa Rican dairy cows under field conditions. Vet. Parasitol. 2004, 123, 149–159, doi:10.1016/j.vetpar.2004.06.016.
[13]
Weston, J.F.; Heuer, C.; Williamson, N.B. Efficacy of a Neospora caninum killed tachyzoite vaccine in preventing abortion and vertical transmission in dairy cattle. Prev. Vet. Med. 2011, 103, 136–144.
[14]
Williams, D.J.; Guy, C.S.; Smith, R.F.; Ellis, J.; Bjorkman, C.; Reichel, M.P.; Trees, A.J. Immunization of cattle with live tachyzoites of Neospora caninum confers protection against fetal death. Infect. Immun. 2007, 75, 1343–1348, doi:10.1128/IAI.00777-06.
[15]
Monney, T.; Debache, K.; Hemphill, A. Vaccines against a major cause of abortion in cattle, Neospora caninum infection. Animals 2011, 1, 306–325, doi:10.3390/ani1030306.
[16]
Hemphill, A.; Vonlaufen, N.; Naguleswaran, A. Cellular and immunological basis of the host-parasite relationship during infection with Neospora caninum. Parasitology 2006, 133, 261–278, doi:10.1017/S0031182006000485.
[17]
Innes, E.A. The host-parasite relationship in pregnant cattle infected with Neospora caninum. Parasitology 2007, 134, 1903–1910, doi:10.1017/S0031182007000194.
[18]
Lopez-Perez, I.C.; Collantes-Fernandez, E.; Aguado-Martinez, A.; Rodriguez-Bertos, A.; Ortega-Mora, L.M. Influence of Neospora caninum infection in BALB/c mice during pregnancy in post-natal development. Vet. Parasitol. 2008, 155, 175–183, doi:10.1016/j.vetpar.2008.05.018.
[19]
Lopez-Perez, I.C.; Risco-Castillo, V.; Collantes-Fernandez, E.; Ortega-Mora, L.M. Comparative effect of Neospora caninum infection in BALB/c mice at three different gestation periods. J. Parasitol. 2006, 92, 1286–1291, doi:10.1645/GE-883R.1.
[20]
Alaeddine, F.; Keller, N.; Leepin, A.; Hemphill, A. Reduced infection and protection from clinical signs of cerebral neosporosis in C57BL/6 mice vaccinated with recombinant microneme antigen NcMIC1. J. Parasitol. 2005, 91, 657–665, doi:10.1645/GE-401R.
[21]
Cannas, A.; Naguleswaran, A.; Muller, N.; Gottstein, B.; Hemphill, A. Reduced cerebral infection of Neospora caninum-infected mice after vaccination with recombinant microneme protein NcMIC3 and ribi adjuvant. J. Parasitol. 2003, 89, 44–50, doi:10.1645/0022-3395(2003)089[0044:RCIONC]2.0.CO;2.
[22]
Debache, K.; Guionaud, C.; Alaeddine, F.; Mevissen, M.; Hemphill, A. Vaccination of mice with recombinant NcROP2 antigen reduces mortality and cerebral infection in mice infected with Neospora caninum tachyzoites. Int. J. Parasitol. 2008, 38, 1455–1463, doi:10.1016/j.ijpara.2008.04.001.
[23]
Debache, K.; Alaeddine, F.; Guionaud, C.; Monney, T.; Muller, J.; Strohbusch, M.; Leib, S.L.; Grandgirard, D.; Hemphill, A. Vaccination with recombinant NcROP2 combined with recombinant NcMIC1 and NcMIC3 reduces cerebral infection and vertical transmission in mice experimentally infected with Neospora caninum tachyzoites. Int. J. Parasitol. 2009, 39, 1373–1384, doi:10.1016/j.ijpara.2009.04.006.
[24]
Monney, T.; Rütti, D.; Schorer, M.; Debache, K.; Grandgirard, D.; Leib, S.L.; Hemphill, A. RecNcMIC3-1-R is a microneme- and rhoptry-based chimeric antigen that protects against acute neosporosis and limits cerebral parasite load in the mouse model for Neospora caninum infection. Vaccine 2011, 29, 6967–6975, doi:10.1016/j.vaccine.2011.07.038.
[25]
Monney, T.; Debache, K.; Grandgirard, D.; Leib, S.L.; Hemphill, A. Vaccination with the recombinant chimeric antigen recNcMIC3-1-R induces a non-protective Th2-type immune response in the pregnant mouse model for N. caninum infection. Vaccine 2012, 30, 6588–6594, doi:10.1016/j.vaccine.2012.08.024.
[26]
Grun, J.L.; Maurer, P.H. Different T helper cell subsets elicited in mice utilizing two different adjuvant vehicles: the role of endogenous interleukin 1 in proliferative responses. Cell. Immunol. 1998, 121, 134–145, doi:10.1016/0008-8749(89)90011-7.
[27]
Feng, X.; Zhang, N.; Tuo, W. Neospora caninum tachyzoite- and antigen-stimulated cytokine production by bone marrow-derived dendritic cells and spleen cells of naive BALB/c Mice. J. Parasitol 2010, 96, 717–723, doi:10.1645/GE-1900.1.
[28]
Rojo-Montejo, S.; Collantes-Fernandez, E.; Regidor-Cerrillo, J.; Rodriguez-Bertos, A.; Prenafeta, A.; Gomez-Bautista, M.; Ortega-Mora, L.M. Influence of adjuvant and antigen dose on protection induced by an inactivated whole vaccine against Neospora caninum infection in mice. Vet. Parasitol. 2011, 175, 220–229, doi:10.1016/j.vetpar.2010.10.028.
[29]
Liddell, S.; Jenkins, M.C.; Collica, C.M.; Dubey, J.P. Prevention of vertical transfer of Neospora caninum in BALB/c mice by vaccination. J. Parasitol. 1999, 85, 1072–1075, doi:10.2307/3285670.
[30]
Yamane, I.; Kitani, H.; Kokuho, T.; Shibahara, T.; Haritani, M.; Hamaoka, T.; Shimizu, S.; Koiwai, M.; Shimura, K.; Yokomizo, Y. The inhibitory effect of interferon gamma and tumor necrosis factor alpha on intracellular multiplication of Neospora caninum in primary bovine brain cells. J. Vet. Med. Sci. 2000, 62, 347–351, doi:10.1292/jvms.62.347.
[31]
Baszler, T.V.; Long, M.T.; McElwain, T.F.; Mathison, B.A. Interferon-gamma and interleukin-12 mediate protection to acute Neospora caninum infection in BALB/c mice. Int. J. Parasitol. 1999, 29, 1635–1646, doi:10.1016/S0020-7519(99)00141-1.
[32]
Bartley, P.M.; Wright, S.E.; Maley, S.W.; Buxton, D.; Nath, M.; Innes, E.A. The development of immune responses in BALB/c mice following inoculation with attenuated or virulent Neospora caninum tachyzoites. Parasite Immunol. 2009, 31, 392–401, doi:10.1111/j.1365-3024.2009.01115.x.
[33]
Debache, K.; Kropf, C.; Schutz, C.A.; Harwood, L.J.; Kauper, P.; Monney, T.; Rossi, N.; Laue, C.; McCullough, K.C.; Hemphill, A. Vaccination of mice with chitosan nanogel-associated recombinant NcPDI against challenge infection with Neospora caninum tachyzoites. Parasite Immunol. 2011, 33, 81–94, doi:10.1111/j.1365-3024.2010.01255.x.
[34]
Nishikawa, Y.; Inoue, N.; Makala, L.; Nagasawa, H. A role for balance of interferon-gamma and interleukin-4 production in protective immunity against Neospora caninum infection. Vet. Parasitol. 2003, 116, 175–184, doi:10.1016/j.vetpar.2003.07.001.
[35]
Kano, R.; Masukata, Y.; Omata, Y.; Kobayashi, Y.; Maeda, R.; Saito, A. Relationship between type 1/type 2 immune responses and occurrence of vertical transmission in BALB/c mice infected with Neospora caninum. Vet. Parasitol. 2005, 129, 159–164, doi:10.1016/j.vetpar.2005.01.004.
[36]
Quinn, H.E.; Miller, C.M.D.; Ellis, J.T. The cell-mediated immune response to Neospora caninum during pregnancy in the mouse is associated with a bias towards production of interleukin-4. Int. J. Parasitol. 2004, 34, 723–732, doi:10.1016/j.ijpara.2004.01.007.
[37]
Baszler, T.V.; McElwain, T.F.; Mathison, B.A. Immunization of BALB/c mice with killed Neospora caninum tachyzoite antigen induces a type 2 immune response and exacerbates encephalitis and neurological disease. Clin. Diagn. Lab. Immunol. 2000, 7, 893–898.
[38]
Long, M.T.; Baszler, T.V.; Mathison, B.A. Comparison of intracerebral parasite load, lesion development, and systemic cytokines in mouse strains infected with Neospora caninum. J. Parasitol. 1998, 84, 316–320, doi:10.2307/3284489.
[39]
Collantes-Fernandez, E.; Arrighi, R.B.; Alvarez-Garcia, G.; Weidner, J.M.; Regidor-Cerrillo, J.; Boothroyd, J.C.; Ortega-Mora, L.M.; Barragan, A. Infected dendritic cells facilitate systemic dissemination and transplacental passage of the obligate intracellular parasite Neospora caninum in mice. PLoS One 2012, 7, e32123, doi:10.1371/journal.pone.0032123.
[40]
Rosbottom, A.; Gibney, E.H.; Guy, C.S.; Kipar, A.; Smith, R.F.; Kaiser, P.; Trees, A.J.; Williams, D.J. Upregulation of cytokines is detected in the placentas of cattle infected with Neospora caninum and is more marked early in gestation when fetal death is observed. Infect. Immun. 2008, 76, 2352–2361, doi:10.1128/IAI.01780-06.
[41]
Aguado-Martinez, A.; Alvarez-Garcia, G.; Fernandez-Garcia, A.; Risco-Castillo, V.; Marugan-Hernandez, V.; Ortega-Mora, L.M. Failure of a vaccine using immunogenic recombinant proteins rNcSAG4 and rNcGRA7 against neosporosis in mice. Vaccine 2009, 27, 7331–7338, doi:10.1016/j.vaccine.2009.09.050.
[42]
Jimenez-Ruiz, E.; Alvarez-Garcia, G.; Aguado-Martinez, A.; Salman, H.; Irache, J.M.; Marugan-Hernandez, V.; Ortega-Mora, L.M. Low efficacy of NcGRA7, NcSAG4, NcBSR4 and NcSRS9 formulated in poly-varepsilon-caprolactone against Neospora caninum infection in mice. Vaccine 2012, 30, 4983–4992, doi:10.1016/j.vaccine.2012.05.033.
[43]
Ribeiro, D.P.; Freitas, M.M.; Cardoso, M.R.; Pajuaba, A.C.; Silva, N.M.; Mineo, T.W.; Silva, J.S.; Mineo, J.R.; Silva, D.A. CpG-ODN combined with Neospora caninum lysate, but not with excreted-secreted antigen, enhances protection against infection in mice. Vaccine 2009, 27, 2570–2579, doi:10.1016/j.vaccine.2009.02.028.
[44]
Rojo-Montejo, S.; Collantes-Fernandez, E.; Rodriguez-Marcos, S.; Perez-Zaballos, F.; Lopez-Perez, I.; Prenafeta, A.; Ortega-Mora, L.M. Comparative efficacy of immunization with inactivated whole tachyzoites versus a tachyzoite-bradyzoite mixture against neosporosis in mice. Parasitology 2011, 138, 1372–1383, doi:10.1017/S0031182011001156.
[45]
Debache, K.; Guionaud, C.; Alaeddine, F.; Hemphill, A. 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. Parasitology 2010, 137, 229–240, doi:10.1017/S0031182009991259.
[46]
Debache, K.; Hemphill, A. Intra-cisternal vaccination induces high-level protection against Neospora caninum infection in mice. Vaccine 2012, 30, 4209–4215, doi:10.1016/j.vaccine.2012.04.050.
[47]
Dubey, J.P.; Carpenter, J.L.; Speer, C.A.; Topper, M.J.; Uggla, A. Newly recognized fatal protozoan disease of dogs. J. Am. Vet. Med. Assoc. 1988, 192, 1269–1285.
[48]
Hemphill, A.; Gottstein, B.; Kaufmann, H. Adhesion and invasion of bovine endothelial cells by Neospora caninum. Parasitology 1996, 112, 183–197, doi:10.1017/S0031182000084754.
[49]
Whitten, M.K. Effect of exteroceptive factors on the oestrous cycle of mice. Nature 1957, 180, 1436, doi:10.1038/1801436a0.
[50]
Müller, N.; Vonlaufen, N.; Gianinazzi, C.; Leib, S.L.; Hemphill, A. Application of real-time fluorescent PCR for quantitative assessment of Neospora caninum infections in organotypic slice cultures of rat central nervous system tissue. J. Clin. Microbiol. 2002, 40, 252–255, doi:10.1128/JCM.40.1.252-255.2002.
[51]
Basto, A.; Piedade, J.; Ramalho, R.; Alves, S.; Soares, H.; Cornelis, P.; Martins, C.; Leitao, A. A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations. J. Biotechnol. 2012, 157, 50–63.