Skip to content
BY-NC-ND 3.0 license Open Access Published by De Gruyter August 4, 2011

Pathological and immunological findings in placentas from pregnant BALB/c mice infected with Neospora caninum at early and late stages of gestation

Inmaculada López-Pérez EMAIL logo , Esther Collantes-Fernández , Silvia Rojo-Montejo , Vanesa Navarro-Lozano , Verónica Risco-Castillo , Valentín Pérez-Pérez , Juana Pereira-Bueno and Luis Ortega-Mora
From the journal Acta Parasitologica

Abstract

Neospora caninum is transmitted from a cow to its foetus by vertical transmission and the timing of infection in gestation is an important factor in determining the disease outcome. Few studies have explored the role of the placenta in the outcome of N. caninum infection during pregnancy. Here, we described the N. caninum presence, parasite load, local immune response, and histopathological lesions at the materno-foetal interface after infection of BALB/c mice at early and late stages of gestation. In mice infected at early gestation, N. caninum DNA was detected in foetoplacentary units 7 days post-infection (PI) and in the placenta, but not in viable foetuses on day 14 PI, indicating that the parasite was multiplying primarily in the placental tissues without reaching the foetus. Moreover, parasite DNA was detected in resorptions, suggesting that foetal death could be a consequence of infection. An increase in IFN-γ, TNF-α and IL-10 expression was observed in N. caninum PCR-positive placentas, which could favour N. caninum foetal transmission and be harmful to both the placenta and the foetus. Histopathological analysis revealed necrosis affecting both the maternal and foetal sides of the placenta. At late gestation, transmission occurred rapidly following infection (day 3 PI), but parasite were rarely found. In addition, an increase in cytokine expression was observed in spleen and placental tissues from infected animals, while a downregulation in IL-4 expression was only observed in the spleen. Finally, necrosis in the placenta was limited to the maternal side, suggesting that the parasite is mainly multiplying in the placental tissue at this stage. Thus, the results of the present study indicate that the placenta may be actively involved in N. caninum pathogenesis.

[1] Abou-Bacar A., Pfaff A.W., Georges S., Letscher-Bru V., Filisetti D., Villard O., Antoni E., Klein J.P., Candolfi E. 2004. Role of NK cells and gamma interferon in transplacental passage of Toxoplasma gondii in a mouse model of primary infection. Infection and Immunity, 72, 1397–1401. DOI: 10.1128/IAI. 72.3.1397-1401.2004. http://dx.doi.org/10.1128/IAI.72.3.1397-1401.200410.1128/IAI.72.3.1397-1401.2004Search in Google Scholar

[2] Amarante-Paffaro A., Queiroz G.S., Corrêa S.T., Spira B., Bevilacqua E. 2004. Phagocytosis as a potential mechanism for microbial defense of mouse placental trophoblast cells. Reproduction, 128, 207–218. DOI: 10.1530/rep.1.00214. http://dx.doi.org/10.1530/rep.1.0021410.1530/rep.1.00214Search in Google Scholar

[3] Barbosa B.F., Silva D.A., Costa I.N., Mineo J.R., Ferro E.A. 2008. BeWo trophoblast cell susceptibility to Toxoplasma gondii is increased by interferon-gamma, interleukin-10 and transforming growth factor-beta 1. Clinical and Experimental Immunology, 151, 536–545. DOI: 10.1111/j.1365-2249.2007.03583.x. http://dx.doi.org/10.1111/j.1365-2249.2007.03583.x10.1111/j.1365-2249.2007.03583.xSearch in Google Scholar

[4] Barr B.C., Rowe J.D., Sverlow S.K., BonDurant R.H., Ardans A.A., Oliver M.N., Conrad P.A. 1994. Experimental reproduction of bovine fetal Neospora infection and death with a bovine Neospora isolate. Journal of Veterinary Diagnostic Investigation, 6, 207–215. http://dx.doi.org/10.1177/10406387940060021210.1177/104063879400600212Search in Google Scholar

[5] Buendía A.J., Sánchez J., Martínez M.C., Cámara P., Navarro J.A., Rodolakis A., Salinas J. 1998. Kinetics of infection and effects on placental cell populations in a murine model of Chlamydia psittaci-induced abortion. Infection and Immunity, 66, 2128–2134. 10.1128/IAI.66.5.2128-2134.1998Search in Google Scholar

[6] Buxton D., Maley S.W., Wright S., Thomson K.M., Rae A.G., Innes E.A. 1998. The pathogenesis of experimental neosporosis in pregnant sheep. Journal of Comparative Pathology, 118, 267–279. DOI: 10.1016/S0021-9975(07)80003-X. http://dx.doi.org/10.1016/S0021-9975(07)80003-X10.1016/S0021-9975(07)80003-XSearch in Google Scholar

[7] Chaouat G., Menu E., Clark D.A., Dy M., Minkowski M., Wegmann T.G. 1990. Control of fetal survival in CBA × DBA/2 mice by lymphokine therapy. Journal of Reproduction and Fertility, 89, 447–458. DOI: 10.1530/jrf.0.0890447. http://dx.doi.org/10.1530/jrf.0.089044710.1530/jrf.0.0890447Search in Google Scholar PubMed

[8] Cole R.A., Lindsay D.S., Blagburn B.L., Dubey J.P. 1995. Vertical transmission of Neospora caninum in mice. Journal of Parasitology, 81, 730–732. DOI: 10.2307/3283962. http://dx.doi.org/10.2307/328396210.2307/3283962Search in Google Scholar

[9] Collantes-Fernández E., RodrÍguez-Bertos A., Arnaiz-Seco I., Moreno B., Aduriz G., Ortega-Mora L.M. 2006. Influence of the stage of pregnancy on Neospora caninum distribution, parasite loads and lesions in aborted bovine foetuses. Theriogenology, 65, 629–641. DOI: 10.1016/j.theriogenology.2005.06.003. http://dx.doi.org/10.1016/j.theriogenology.2005.06.00310.1016/j.theriogenology.2005.06.003Search in Google Scholar PubMed

[10] Collantes-Fernández E., Zaballos Á., Álvarez-García G., Ortega-Mora L.M. 2002. Quantitative detection of Neospora caninum in bovine aborted fetuses and experimentally infected mice by real-time PCR. Journal of Clinical Microbiology, 40, 1194–1198. DOI: 10.1128/JCM.40.4.1194-1198.2002. http://dx.doi.org/10.1128/JCM.40.4.1194-1198.200210.1128/JCM.40.4.1194-1198.2002Search in Google Scholar PubMed PubMed Central

[11] Dubey J.P., Buxton D., Wouda W. 2006. Pathogenesis of bovine neosporosis. Journal of Comparative Pathology, 134, 267–289. DOI: 10.1016/j.jcpa.2005.11.004. http://dx.doi.org/10.1016/j.jcpa.2005.11.00410.1016/j.jcpa.2005.11.004Search in Google Scholar

[12] Dubey J.P., Lindsay D.S. 1996. A review of Neospora caninum and neosporosis. Veterinary Parasitology, 67, 1–59. DOI: 10.1016/S0304-4017(96)01035-7. http://dx.doi.org/10.1016/S0304-4017(96)01035-710.1016/S0304-4017(96)01035-7Search in Google Scholar

[13] Dubey J.P., Lindsay D.S., Anderson M.L., Davis S.W., Shen S.K. 1992. Induced transplacental transmission of Neospora caninum in cattle. Journal of the American Veterinary Medical Association, 201, 709–713. Search in Google Scholar

[14] Eperon S., Brönnimann K., Hemphill A., Gottstein B. 1999. Susceptibility of B-cell deficient C57BL/6 (μMT) mice to Neospora caninum infection. Parasite Immunology, 21, 225–236. DOI: 10.1046/j.1365-3024.1999.00223.x. http://dx.doi.org/10.1046/j.1365-3024.1999.00223.x10.1046/j.1365-3024.1999.00223.xSearch in Google Scholar

[15] Ferro E.A., Silva D.A., Bevilacqua E., Mineo J.R. 2002. Effect of Toxoplasma gondii infection kinetics on trophoblast cell population in Calomys callosus, a model of congenital toxoplasmosis. Infection and Immunity, 70, 7089–7094. DOI: 10.1128/IAI.70.12.7089-7094.2002. http://dx.doi.org/10.1128/IAI.70.12.7089-7094.200210.1128/IAI.70.12.7089-7094.2002Search in Google Scholar

[16] Innes E.A., Andrianarivo A.G., Björkman C., Williams D.J., Conrad P.A. 2002. Immune responses to Neospora caninum and prospects for vaccination. Trends in Parasitology, 18, 497–504. DOI: 10.1016/S1471-4922(02)02372-3. http://dx.doi.org/10.1016/S1471-4922(02)02372-310.1016/S1471-4922(02)02372-3Search in Google Scholar

[17] Innes E.A., Wright S.E., Maley S., Rae A., Schock A., Kirvar E., Bartley P., Hamilton C., Carey I.M., Buxton D. 2001. Protection against vertical transmission in bovine neosporosis. International Journal for Parasitology, 31, 1523–1534. DOI: 10.1016/S0020-7519(01)00284-3. http://dx.doi.org/10.1016/S0020-7519(01)00284-310.1016/S0020-7519(01)00284-3Search in Google Scholar

[18] Kano R., Masukata Y., Omata Y., Kobayashi Y., Maeda R., Saito A. 2005. Relationship between type 1/type 2 immune responses and occurrence of vertical transmission in BALB/c mice infected with Neospora caninum. Veterinary Parasitology, 129, 159–164. DOI: 10.1016/j.vetpar.2005.01.004. http://dx.doi.org/10.1016/j.vetpar.2005.01.00410.1016/j.vetpar.2005.01.004Search in Google Scholar PubMed

[19] Krishnan L., Guilbert L.J., Wegmann T.G., Belosevic M., Mosmann T.R. 1996. T helper 1 response against Leishmania major in pregnant C57BL/6 mice increases implantation failure and fetal resorptions. Correlation with increased IFN-gamma and TNF and reduced IL-10 production by placental cells. Journal of Immunology, 156, 653–662. Search in Google Scholar

[20] Liddell S., Jenkins M.C., Dubey J.P. 1999. Vertical transmission of Neospora caninum in BALB/c mice determined by polymerase chain reaction detection. Journal of Parasitology, 85, 550–555. DOI: 10.2307/3285794. 10.2307/3285794Search in Google Scholar

[21] Lin H., Mosmann T.R., Guilbert L., Tuntipopipat S., Wegmann T.G. 1993. Synthesis of T helper 2-type cytokines at the maternalfetal interface. Journal of Immunology, 151, 4562–4573. Search in Google Scholar

[22] Livak K., Schmittgen T.D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 25, 402–408. DOI: 10.1006/meth.2001.1262. http://dx.doi.org/10.1006/meth.2001.126210.1006/meth.2001.1262Search in Google Scholar PubMed

[23] Long M.T., Baszler T.V. 1996. Fetal loss in BALB/c mice infected with Neospora caninum. Journal of Parasitology, 82, 608–611. DOI: 10.2307/3283785. http://dx.doi.org/10.2307/328378510.2307/3283785Search in Google Scholar

[24] Long M.T., Baszler T.V. 2000. Neutralization of maternal IL-4 modulates congenital protozoal transmission: comparison of innate versus acquired immune responses. Journal of Immunology, 164, 4768–4774. Search in Google Scholar

[25] López-Pérez I.C., Collantes-Fernández E., Aguado-MartÍnez A., Rodríguez-Bertos A., Ortega-Mora L.M. 2008. Influence of Neospora caninum infection in BALB/c mice during preg nancy in post-natal development. Veterinary Parasitology, 155, 175–183. DOI: 10.1016/j.vetpar.2008.05.018. http://dx.doi.org/10.1016/j.vetpar.2008.05.01810.1016/j.vetpar.2008.05.018Search in Google Scholar

[26] López-Pérez I.C., Collantes-Fernández E., Rojo-Montejo S., Navarro-Lozano V., Risco-Castillo V., Pérez-Pérez V., Pereira-Bueno J., Ortega-Mora L.M. 2010. Effects of Neospora caninum infection at mid-gestation on placenta in a pregnant mouse model. Journal of Parasitology, 96, 1017–1020. DOI: 10.1645/GE-2347.1. http://dx.doi.org/10.1645/GE-2347.110.1645/GE-2347.1Search in Google Scholar

[27] López-Pérez I., Risco-Castillo V., Collantes-Fernández E., Ortega-Mora L.M. 2006. Comparative effect of infection with Neospora caninum at the three gestation periods. Journal of Parasitology, 92, 1286–1291. DOI: 10.1645/GE-883R.1. http://dx.doi.org/10.1645/GE-883R.110.1645/GE-883R.1Search in Google Scholar

[28] Macaldowie C., Maley S.W., Wright S., Bartley P., Esteban-Redondo I., Buxton D., Innes E.A. 2004. Placental pathology associated with fetal death in cattle inoculated with Neospora caninum by two different routes in early pregnancy. Journal of Comparative Pathology, 131, 142–156. DOI: 10.1016/j.jcpa. 2004.02.005. http://dx.doi.org/10.1016/j.jcpa.2004.02.00510.1016/j.jcpa.2004.02.005Search in Google Scholar

[29] Maley S.W., Buxton D., Macaldowie C.N., Anderson I.E., Wright S.E., Bartley P.M., Esteban-Redondo I., Hamilton C.M., Storset A.K., Innes E.A. 2006. Characterization of the immune response in the placenta of cattle experimentally infected with Neospora caninum in early gestation. Journal of Comparative Pathology, 135, 130–141. DOI: 10.1016/j.jcpa. 2006.07.001 http://dx.doi.org/10.1016/j.jcpa.2006.07.00110.1016/j.jcpa.2006.07.001Search in Google Scholar

[30] Maley S.W., Buxton D., Rae A.G., Wright S.E., Schock A., Bartley P.M., Esteban-Redondo I., Swales C., Hamilton C.M., Sales J., Innes E.A. 2003. The pathogenesis of neosporosis in pregnant cattle: inoculation at mid-gestation. Journal of Comparative Pathology, 129, 186–195. DOI: 10.1016/S0021-9975 (03)00032-X. http://dx.doi.org/10.1016/S0021-9975(03)00032-X10.1016/S0021-9975(03)00032-XSearch in Google Scholar

[31] Moormann A.M., Sullivan D., Rochford R.A., Chensue S.W., Bock P.J., Nyirenda T., Meshnick S.R. 1999. Malaria and pregnancy: placental cytokine expression and its relationship to intrauterine growth retardation. Journal of Infectious Diseases, 180, 1987–1993. DOI: 10.1086/315135. http://dx.doi.org/10.1086/31513510.1086/315135Search in Google Scholar

[32] Morrison D. 2002. How to improve statistical analysis in parasitology research publications. International Journal for Parasitology, 32, 1065–1070. DOI:10.1016/S0020-7519(02)000 64-4. http://dx.doi.org/10.1016/S0020-7519(02)00064-410.1016/S0020-7519(02)00064-4Search in Google Scholar

[33] Ohmori Y., Hamilton T.A. 1997. IL-4-induced STAT6 suppresses IFN-gamma-stimulated STAT1-dependent transcription in mouse macrophages. Journal of Immunology, 159, 5474–5482. Search in Google Scholar

[34] Omata Y., Nidaira M., Kano R., Kobayashi Y., Koyama T., Furuoka H., Maeda R., Matsui T., Saito A. 2004. Vertical transmission of Neospora caninum in BALB/c mice in both acute and chronic infection. Veterinary Parasitology, 121, 323–328. DOI: 10.1016/j.vetpar.2004.03.003. http://dx.doi.org/10.1016/j.vetpar.2004.03.00310.1016/j.vetpar.2004.03.003Search in Google Scholar

[35] Pereira-Bueno J., Quintanilla-Gozalo A., Pérez-Pérez V., Espi-Felgueroso A., álvarez-GarcÍa G., Collantes-Fernández E., Ortega-Mora L.M. 2003. Evaluation by different diagnostic techniques of bovine abortion associated with Neospora caninum in Spain. Veterinary Parasitology, 111, 143–152. DOI: 10.1016/S0304-4017(02)00361-8. http://dx.doi.org/10.1016/S0304-4017(02)00361-810.1016/S0304-4017(02)00361-8Search in Google Scholar

[36] Pérez-Zaballos F.J., Ortega-Mora L.M., álvarez-GarcÍa G., Collantes-Fernández E., Navarro-Lozano V., GarcÍa-Villada L., Costas E. 2005. Adaptation of Neospora caninum isolates to cell-culture changes: an argument in favor of its clonal population structure. Journal of Parasitology, 91, 507–510. DOI: 10.1645/GE-381R1. http://dx.doi.org/10.1645/GE-381R110.1645/GE-381R1Search in Google Scholar

[37] Pfaff A.W., Villard O., Klein J.P., Mousli M., Candolfi E. 2005. Regulation of Toxoplasma gondii multiplication in BeWo trophoblast cells: cross-regulation of nitric oxide production and polyamine biosynthesis. International Journal for Parasitology, 35, 1569–1576. DOI: 10.1016/j.ijpara.2005.08.003. http://dx.doi.org/10.1016/j.ijpara.2005.08.00310.1016/j.ijpara.2005.08.003Search in Google Scholar

[38] Quinn H.E., Ellis J.T., Smith N.C. 2002a. Neospora caninum: a cause of immune-mediated failure of pregnancy? Trends in Parasitology, 18, 391–394. DOI: 10.1016/S1471-4922(02)02324-3. http://dx.doi.org/10.1016/S1471-4922(02)02324-310.1016/S1471-4922(02)02324-3Search in Google Scholar

[39] Quinn H.E., Miller C.M., Ellis J.T. 2004. The cell-mediated immune response to Neospora caninum during pregnancy in the mouse is associated with a bias towards production of interleukin-4. International Journal for Parasitology, 34, 723–732. DOI: 10.1016/j.ijpara.2004.01.007. http://dx.doi.org/10.1016/j.ijpara.2004.01.00710.1016/j.ijpara.2004.01.007Search in Google Scholar

[40] Quinn H.E., Miller C.M., Ryce C., Windsor P.A., Ellis J.T. 2002b. Characterization of an outbred pregnant mouse model of Neospora caninum infection. Journal of Parasitology, 88, 691–696. DOI: 10.2307/3285344. http://dx.doi.org/10.1645/0022-3395(2002)088[0691:COAOPM]2.0.CO;2Search in Google Scholar

[41] Raghupathy R. 1997. Th1-type immunity is incompatible with successful pregnancy. Immunology Today, 18, 478–482. DOI: 10.1016/S0167-5699(97)01127-4. http://dx.doi.org/10.1016/S0167-5699(97)01127-410.1016/S0167-5699(97)01127-4Search in Google Scholar

[42] Rosbottom A., Gibney E.H., Guy C.S., Kipar A., Smith R.F., Kaiser P., Trees A.J., Williams D.J. 2008. An 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. Infection and Immunity, 76, 2352–2361. DOI: 10.1128/IAI.01780-06. http://dx.doi.org/10.1128/IAI.01780-0610.1128/IAI.01780-06Search in Google Scholar

[43] Rosbottom A., Gibney E.H., Kaiser C., Smith R.F., Robinson R., Kipar A., Williams D.J. 2011. Up regulation of the maternal immune response in the placenta of cattle naturally infected with Neospora caninum. PLoS One, 6, e15799. DOI: 10.1371/journal.pone.0015799. http://dx.doi.org/10.1371/journal.pone.001579910.1371/journal.pone.0015799Search in Google Scholar

[44] Trees A.J., Williams D.J.L. 2005. Endogenous and exogenous transplacental infection in Neospora caninum and Toxoplasma gondii. Trends in Parasitology, 21, 558–561. DOI: 10.1016/j.pt.2005.09.005. http://dx.doi.org/10.1016/j.pt.2005.09.00510.1016/j.pt.2005.09.005Search in Google Scholar

[45] Varona R., Cadenas V., Gómez L., MartÍnez A., Márquez G. 2005. CCR6 regulates CD4+ T-cell-mediated acute graft-versushost disease responses. Blood, 106, 18–26. DOI: 10.1182/blood-2004-08-2996. http://dx.doi.org/10.1182/blood-2004-08-299610.1182/blood-2004-08-2996Search in Google Scholar

[46] Wegmann T.G., Lin H., Guilbert L., Mosmann T.R. 1993. Bidirectional cytokine interactions in the maternal-fetal relationship: is successful pregnancy a Th2 phenomenon? Immunology Today, 14, 353–356. DOI: 10.1016/0167-5699(93)90235-D. http://dx.doi.org/10.1016/0167-5699(93)90235-D10.1016/0167-5699(93)90235-DSearch in Google Scholar

[47] Whitten M.K. 1957. Effect of exteroceptive factors on the oestrous cycle of mice. Nature, 180(4599): 1436. DOI: 10.1038/1801436a0. http://dx.doi.org/10.1038/1801436a010.1038/1801436a0Search in Google Scholar

[48] Williams D.J., Guy C.S., McGarry J.W., Guy F., Tasker L., Smith R.F., MacEachern K., Cripps P.J., Kelly D.F., Trees A.J. 2000. Neospora caninum-associated abortion in cattle: the time of experimentally-induced parasitaemia during gestation determines foetal survival. Parasitology, 121, 347–358. DOI: 10.1017/S0031182099006587. http://dx.doi.org/10.1017/S003118209900658710.1017/S0031182099006587Search in Google Scholar

[49] Williams D.J., Guy C.S., Smith R.F., Guy F., McGarry J.W., McKay J.S., Trees A.J. 2003. First demonstration of protective immunity against foetopathy in cattle with latent Neospora caninum infection. International Journal for Parasitology, 33, 1059–1065. DOI: 10.1016/S0020-7519(03)00143-7. http://dx.doi.org/10.1016/S0020-7519(03)00143-710.1016/S0020-7519(03)00143-7Search in Google Scholar

Published Online: 2011-8-4
Published in Print: 2011-9-1

© 2011 W. Stefański Institute of Parasitology, PAS

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Downloaded on 1.12.2022 from frontend.live.degruyter.dgbricks.com/document/doi/10.2478/s11686-011-0052-8/html
Scroll Up Arrow