Transfection of live, tick derived sporozoites of the protozoan Apicomplexan parasite Theileria parva

Ine De Goeyse, Famke Jansen, Maxime Madder, Kyoko Hayashida, Dirk Berkvens, Dirk Dobbelaere, Dirk Geysen

    Research output: Contribution to journalA1: Web of Science-article

    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.

    Original languageEnglish
    JournalVeterinary Parasitology
    Volume208
    Issue number3-4
    Pages (from-to)238-241
    Number of pages4
    ISSN0304-4017
    DOIs
    Publication statusPublished - 2015

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