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Infection and Immunity, January 2004, p. 576-583, Vol. 72, No. 1
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.1.576-583.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

In Vitro Induction of Neospora caninum Bradyzoites in Vero Cells Reveals Differential Antigen Expression, Localization, and Host-Cell Recognition of Tachyzoites and Bradyzoites

Nathalie Vonlaufen,1* Nicole Guetg,1 Arunasalam Naguleswaran,1 Norbert Müller,1 Camilla Björkman,2 Gereon Schares,3 Daniela von Blumroeder,3 John Ellis,4 and Andrew Hemphill1*

Institute of Parasitology, Faculties of Veterinary Medicine and Medicine, University of Berne, CH-3012 Berne, Switzerland,1 Swedish University of Agricultural Sciences, Department of Ruminant Medicine and Veterinary Epidemiology and Department of Parasitology, Uppsala, Sweden,2 Federal Research Centre for Virus Diseases of Animals, D-16868 Wusterhausen, Germany,3 Department of Cell and Molecular Biology, University of Technology, Sydney, NSW 2007, Australia4

Received 11 July 2003/ Returned for modification 16 August 2003/ Accepted 12 September 2003

We report on an optimized method for the in vitro culture of tissue cyst-forming Neospora caninum bradyzoites in Vero cells and the separation of viable parasites from host cells. Treatment of tachyzoite-infected Vero cell cultures with 17 µM sodium nitroprusside for 8 days severely scaled down parasite proliferation, led to reduced expression of tachyzoite surface antigens, and induced the expression of the bradyzoite marker NcBAG1 and the cyst wall antigen recognized by the monoclonal antibody MAbCC2. Transmission electron microscopy demonstrated that intracellular parasites were located within parasitophorous vacuoles that were surrounded by a cyst wall-like structure, and the dense granule antigens NcGRA1, NcGRA2, and NcGRA7 were incorporated into the cyst wall. Adhesion-invasion assays employing purified tachyzoites and bradyzoites showed that tachyzoites adhered to, and invaded, Vero cells with higher efficiency than bradyzoites. However, removal of terminal sialic acid residues from either the host cell or the parasite surface increased the invasion of Vero cells by bradyzoites, but not tachyzoites.

* Corresponding author. Mailing address: Institute of Parasitology, University of Berne, Länggass-Strasse 122, CH-3012 Bern, Switzerland. Phone: 41 31 6312384. Fax: 41 31 6312477. E-mail for Andrew Hemphill: hemphill{at}ipa.unibe.ch. E-mail for Nathalie Vonlaufen: vonlaufen{at}ipa.unibe.ch.

Editor: W. A. Petri, Jr.

Infection and Immunity, January 2004, p. 576-583, Vol. 72, No. 1
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.1.576-583.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Keller, N., Riesen, M., Naguleswaran, A., Vonlaufen, N., Stettler, R., Leepin, A., Wastling, J. M., Hemphill, A. (2004). Identification and Characterization of a Neospora caninum Microneme-Associated Protein (NcMIC4) That Exhibits Unique Lactose-Binding Properties. Infect. Immun. 72: 4791-4800 [Abstract] [Full Text]  


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