title: Transfection of live, tick derived sporozoites of the protozoan Apicomplexan parasite Theileria parva.
abstract: Theileria parva is an important veterinary protozoan causing the tick-borne disease East Coast fever. Transfection of Theileria parasites will facilitate the investigation of many aspects of this apicomplexan infection and its unique host-parasite interaction. The pathogen has the extraordinary capacity of transforming B and T cells into clonally dividing cancerous cell lines in a reversible way. Sequence data of the entire T. parva genome are available and in vitro infected cell lines can easily be generated, thereby eliminating the use of animals in the evaluation of the evolution of the transfected sporozoites. Here we report, for the first time, on experiments towards successful transient transfection of T. parva sporozoites, making use of a new generation transfection device. Plasmid DNA containing the strong EF-1α promoter and an Azami Green reporter gene were integrated by nucleofection into freshly purified T. parva sporozoites. Expression of Azami Green was detected with a fluorescence microscope and confirmed by counter staining with a monoclonal directed against a sporozoite protein. Despite the fact that transfection efficiencies are still low, this is the first step towards a stable infection method of T. parva parasites. In the long run, transfected parasites might become an alternative way to induce immunity without clinical signs.
authors: Institute of Tropical Medicine, Department of Biomedical Sciences, Nationalestraat 155, 2000 Antwerp, Belgium. Electronic address: idegoeyse@itg.be.
authors: Ine
authors: I
authors: De Goeyse
authors: Institute of Tropical Medicine, Department of Biomedical Sciences, Nationalestraat 155, 2000 Antwerp, Belgium.
authors: Famke
authors: F
authors: Jansen
authors: Institute of Tropical Medicine, Department of Biomedical Sciences, Nationalestraat 155, 2000 Antwerp, Belgium.
authors: Maxime
authors: M
authors: Madder
authors: Division of Collaboration and Education, Research Center for Zoonosis Control, Hokkaido University, Sapporo-shi, Hokkaido 001-0020, Japan.
authors: Kyoko
authors: K
authors: Hayashida
authors: Institute of Tropical Medicine, Department of Biomedical Sciences, Nationalestraat 155, 2000 Antwerp, Belgium.
authors: Dirk
authors: D
authors: Berkvens
authors: Division of Molecular Pathobiology, Vetsuisse Faculty, University of Bern, Länggassstrasse 122, CH-3012 Bern, Switzerland.
authors: Dirk
authors: D
authors: Dobbelaere
authors: Institute of Tropical Medicine, Department of Biomedical Sciences, Nationalestraat 155, 2000 Antwerp, Belgium.
authors: Dirk
authors: D
authors: Geysen
mesh_terms: D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D034101:Sporozoites / Q000502:physiology*; D016797:Theileria parva / Q000235:genetics / Q000502:physiology*; D014162:Transfection
chemical_list: D049452:Green Fluorescent Proteins
keywords: Nucleofection; Sporozoites; Theileria parva; Transfection