In Vitro Induction of Neospora caninum Bradyzoites in Vero Cells Reveals Differential Antigen Expression, Localization, and Host-Cell Recognition of Tachyzoites and Bradyzoites
- 1 January 2004
- journal article
- research article
- Published by American Society for Microbiology in Infection and Immunity
- Vol. 72 (1), 576-583
- https://doi.org/10.1128/iai.72.1.576-583.2004
Abstract
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.Keywords
This publication has 73 references indexed in Scilit:
- Tissue Culture and Explant Approaches to Studying and VisualizingNeospora caninumand Its Interactions with the Host CellMicroscopy and Microanalysis, 2004
- Neospora caninum and Toxoplasma gondii: a novel adhesion/invasion assay reveals distinct differences in tachyzoite–host cell interactionsExperimental Parasitology, 2003
- Isolation of Neospora caninum genes detected during a chronic murine infectionInternational Journal for Parasitology, 2001
- The Role of Mucins in Host–Parasite Interactions. Part I – Protozoan ParasitesParasitology Today, 2000
- Use of purified tachyzoite surface antigen p38 in an ELISA to diagnose bovine neosporosisInternational Journal for Parasitology, 2000
- Long-Term Culture of Murine Epidermal KeratinocytesJournal of Investigative Dermatology, 2000
- Comparative ultrastructure of tachyzoites, bradyzoites, and tissue cysts of Neospora caninum and Toxoplasma gondiiInternational Journal for Parasitology, 1999
- Isolation of the cDNA encoding a dense granule associated antigen (NCDG2) of Neospora caninum1Note: Nucleotide sequence data reported in this paper are available from the GenBank™ database under accession number AF029350.1Molecular and Biochemical Parasitology, 1998
- Identification and characterisation of a dense granuleassociated protein in Neospora caninum tachyzoitesInternational Journal for Parasitology, 1998
- Characterization of the first European isolate ofNeospora caninum(Dubey, Carpenter, Speer, Topper and Uggla)Parasitology, 1995