In vitro maturation of Toxoplasma gondii bradyzoites in human myotubes and their metabolomic characterization

Céline Christiansen, Deborah Maus, Ellen Hoppenz, Mateo Murillo-León, Tobias Hoffmann, Jana Scholz, Florian Melerowicz, Tobias Steinfeldt, Frank Seeber and Martin Blume ()
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Céline Christiansen: Robert Koch-Institute
Deborah Maus: Robert Koch-Institute
Ellen Hoppenz: Robert Koch-Institute
Mateo Murillo-León: Medical Center University of Freiburg
Tobias Hoffmann: Centre for Biological Threats and Special Pathogens 4, Robert Koch-Institute
Jana Scholz: Robert Koch-Institute
Florian Melerowicz: Robert Koch-Institute
Tobias Steinfeldt: Medical Center University of Freiburg
Frank Seeber: Robert Koch-Institute
Martin Blume: Robert Koch-Institute

Nature Communications, 2022, vol. 13, issue 1, 1-15

Abstract: Abstract The apicomplexan parasite Toxoplasma gondii forms bradyzoite-containing tissue cysts that cause chronic and drug-tolerant infections. However, current in vitro models do not allow long-term culture of these cysts to maturity. Here, we developed a human myotube-based in vitro culture model of functionally mature tissue cysts that are orally infectious to mice and tolerate exposure to a range of antibiotics and temperature stresses. Metabolomic characterization of purified cysts reveals global changes that comprise increased levels of amino acids and decreased abundance of nucleobase- and tricarboxylic acid cycle-associated metabolites. In contrast to fast replicating tachyzoite forms of T. gondii these tissue cysts tolerate exposure to the aconitase inhibitor sodium fluoroacetate. Direct access to persistent stages of T. gondii under defined cell culture conditions will be essential for the dissection of functionally important host-parasite interactions and drug evasion mechanisms. It will also facilitate the identification of new strategies for therapeutic intervention.

Date: 2022
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DOI: 10.1038/s41467-022-28730-w

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