Toxoplasma gondii: Bradyzoite Differentiation In Vitro and In Vivo

Joshua Mayoral, Manlio Di Cristina, Vern B. Carruthers, Louis M. Weiss

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

Toxoplasma gondii, a member of the Apicomplexa, is known for its ability to infect an impressive range of host species. It is a common human infection that causes significant morbidity in congenitally infected children and immunocompromised patients. This parasite can be transmitted by bradyzoites, a slowly replicating life stage found within intracellular tissue cysts, and oocysts, the sexual life cycle stage that develops in domestic cats and other Felidae. T. gondii bradyzoites retain the capacity to revert back to the quickly replicating tachyzoite life stage, and when the host is immune compromised unrestricted replication can lead to significant tissue destruction. Bradyzoites are refractory to currently available Toxoplasma treatments. Improving our understanding of bradyzoite biology is critical for the development of therapeutic strategies to eliminate latent infection. This chapter describes a commonly used protocol for the differentiation of T. gondii tachyzoites into bradyzoites using human foreskin fibroblast cultures and a CO2-limited alkaline cell media, which results in a high proportion of differentiated bradyzoites for further study. Also described are methods for purifying tissue cysts from chronically infected mouse brain using isopycnic centrifugation and a recently developed approach for measuring bradyzoite viability.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages269-282
Number of pages14
DOIs
StatePublished - Jan 1 2020

Publication series

NameMethods in Molecular Biology
Volume2071
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

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Keywords

  • Bradyzoite
  • Differentiation
  • In vitro
  • In vivo
  • Stress
  • Tissue cyst
  • Toxoplasma

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Mayoral, J., Di Cristina, M., Carruthers, V. B., & Weiss, L. M. (2020). Toxoplasma gondii: Bradyzoite Differentiation In Vitro and In Vivo. In Methods in Molecular Biology (pp. 269-282). (Methods in Molecular Biology; Vol. 2071). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9857-9_15