Interaction and cystogenesis of Toxoplasma gondii within skeletal muscle cells in vitro

Guimarães, Erick Vaz; Carvalho, Laís de; Barbosa, Helene Santos

doi:10.1590/s0074-02762009000200007

Cited by 15 publications

(9 citation statements)

References 30 publications

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“…Parasites of T. gondii ME49 strain are routinely maintained in C57BL/6 mice. Tissue cysts are obtained from the brains of mice inoculated intraperitoneally with 50 cysts/animal, as described elsewhere [38]. Briefly, cysts were ruptured with acid pepsin solution and free parasites were added to Vero cells cultures.…”

Section: Parasites Culturementioning

confidence: 99%

Extracellular Traps Released by Neutrophils from Cats are Detrimental to Toxoplasma gondii Infectivity

et al. 2020

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Toxoplasma gondii is the causative agent of toxoplasmosis, an infectious disease that affects over 30% of the human world population, causing fatal infections in immunocompromised individuals and neonates. The life cycle of T. gondii is complex, and involves intermediate hosts (birds and mammals) and definitive hosts (felines, including domestic cats). The innate immune repertoire against the parasite involves the production of neutrophil extracellular traps (NET), and neutrophils from several intermediate hosts produce NET induced by T. gondii. However, the mechanisms underlying NET release in response to the parasite have been poorly explored. Therefore, the aims of this study were to investigate whether neutrophils from cats produce NET triggered by T. gondii and to understand the mechanisms thereby involved. Neutrophils from cats were stimulated with T. gondii tachyzoites and NET-derived DNA in the supernatant was quantified during the time. The presence of histone H1 and myeloperoxidase was detected by immunofluorescence. We observed that cat neutrophils produce both classical and rapid/early NET stimulated by T. gondii. Inhibition of elastase, intracellular calcium, and phosphatidylinositol 3-kinase (PI3K)-δ partially blocked classical NET release in response to the parasite. Electron microscopy revealed strands and networks of DNA in close contact or completely entrapping parasites. Live imaging showed that tachyzoites are killed by NET. We conclude that the production of NET is a conserved strategy to control infection by T. gondii amongst intermediate and definitive hosts.

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Section: Parasites Culturementioning

confidence: 99%

Extracellular Traps Released by Neutrophils from Cats are Detrimental to Toxoplasma gondii Infectivity

et al. 2020

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“…In contrast, in vitro models are scalable and the stage conversion of T. gondii can be induced by a variety of treatments, such as alkaline, heat and chemical stress 23 , nutrient starvation 24 , excess adenosine 25 , inhibitors of parasite protein kinases 26 as well as host cell-dependent factors such as cell cycle arrest and low glycolytic flux 20 , 27 , 28 . In addition, the infection of C2C12 murine skeletal muscle cells, neurons 29 , 30 , and primary murine brain cells 31 facilitates spontaneous stage conversion of cystogenic strains. Generally, maturation times of in vitro cysts remain brief, but cysts develop detectable tolerance against short exposure to low doses of pyrimethamine after three days 10 and become orally infectious to mice at high doses after five days of culture 32 .…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2022

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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.

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“…In contrast, in vitro models are scalable and the stage conversion of T. gondii can be induced by a variety of treatments, such as alkaline, heat and chemical stress (Soete et al, 1994), nutrient starvation (Fox et al, 2004), excess adenosine (Mahamed et al, 2012), inhibitors of parasite protein kinases (Donald et al, 2002) as well as host cell-dependent factors such as cell cycle arrest and low glycolytic flux (Lüder and Rahman, 2017; Mayoral et al, 2020; Weilhammer et al, 2012). In addition, the infection of C2C12 murine skeletal muscle cells and neurons (Guimarães et al, 2009; Guimaraes et al, 2008) facilitates spontaneous stage conversion of cystogenic strains. Generally, maturation times of cysts in vitro -cysts remain brief but cysts develop detectable tolerance against short exposure to low doses of pyrimethamine after three days (Murata et al, 2017) and become orally infectious to mice at high doses after five days of culture (Fux et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

In vitromaturation ofToxoplasma gondiibradyzoites in human myotubes and their metabolomic characterization

Christiansen

Maus

Melerowicz

et al. 2021

Preprint

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SummaryThe apicomplexan parasite Toxoplasma gondii causes chronic and drug-tolerant infections, yet current models do not permit metabolomic characterization of these persisting tissue cysts. Here, we developed a human myotube-based in vitro culture model of functionally mature tissue cysts that enabled direct measurements of their metabolome. The cysts are functionally mature and tolerate exposure to a range of antibiotics and to extended temperature stresses and are orally infectious to mice. Metabolomic characterization of purified cysts reveals global changes that comprise systematically increased levels of amino acids and decrease abundance of nucleobase- and tricarboxylic acid cycle-associated metabolites. Consequently, pharmacological modulation of the TCA cycle in T. gondii bradyzoites reveals that this pathway is rendered dispensable during parasite stage conversion. Direct access to persisting parasite stages will be essential for the dissection of functionally important host-parasite interactions and drug evasion mechanisms and also help to identify new strategies for therapeutic intervention.Highlights–Toxoplasma gondii forms mature tissue cysts in immortalized human myotubes–In vitro cysts of T. gondii develop drug tolerance and temperature stress resistance–Untargeted metabolomic characterization of tissue cysts reveals a distinct metabolome–The mitochondrial tricarboxylic acid cycle is dispensable in T. gondii bradyzoites

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Interaction and cystogenesis of Toxoplasma gondii within skeletal muscle cells in vitro

Abstract: Infection by the protozoan parasite

Cited by 15 publications

References 30 publications

Extracellular Traps Released by Neutrophils from Cats are Detrimental to Toxoplasma gondii Infectivity

Extracellular Traps Released by Neutrophils from Cats are Detrimental to Toxoplasma gondii Infectivity

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

In vitromaturation ofToxoplasma gondiibradyzoites in human myotubes and their metabolomic characterization

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