Animal Models for Toxoplasma gondii Infection

Abstract

AbstractToxoplasma gondii is an obligate intracellular protozoan parasite that commonly infects mammals and birds throughout the world. This protocol describes murine models of acute T. gondii infection, toxoplasmic encephalitis and toxoplasma retinochoroiditis. T. gondii infection in severe combined immunodeficient (SCID) mice, deficient in T and B cells, has allowed for the study of T cell‐independent mechanisms of defense against intracellular organisms, as described here. The uracil auxotroph strain cps1‐1 and temperature‐sensitive mutant strains of T. gondii induce protection against challenge with virulent strains of the parasite. They have allowed studies of immunization and adoptive‐transfer experiments. A protocol is provided for infection with these mutant strains. The EGS strain of T. gondii has the unique feature of spontaneously forming tissue cysts in cell culture. Dual fluorescent reporter stains of this strain have allowed the study of tachyzoite to bradyzoite transitions in vitro and in vivo. A protocol for in vitro and in vivo growth of this strain and tissue cyst isolation is provided. Genetic manipulation of T. gondii and mice has led to the development of parasites that express fluorescent proteins as well as mice with fluorescently labeled leukocytes. This together with the use of T. gondii that express model antigens and transgenic mice that express the appropriate T cell receptor have facilitated the in vivo study of parasite host‐interaction. In addition, parasites that express bioluminescent markers have made it possible to study the dynamics of infection in real time using bioluminescence imaging. Support protocols present methodology for evaluation of progression of infection and immune response to the parasite that includes these newer methodologies. In addition, support protocols address the maintenance of T. gondii tissue cysts and tachyzoites, as well as preparation of T. gondii lysate antigens. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.Basic Protocol 1: Induction of acute T. gondii infection in miceBasic Protocol 2: Model of toxoplasmic encephalitis and toxoplasma retinochoroiditis in chronically infected miceBasic Protocol 3: Assessment of T. gondii invasion into neural tissueBasic Protocol 4: T. gondii infection in scid/scid (SCID) miceBasic Protocol 5: Infection with the uracil auxotroph strain CPS1‐1 or the temperature‐sensitive TS‐4 strain of T. gondiiBasic Protocol 6: In vivo and in vitro maintenance of the EGS strain of T. gondiiSupport Protocol 1: Assessment of progression of infection and immune response to T. gondiiSupport Protocol 2: Maintenance of a bank of T. gondii cysts of the ME49 strainSupport Protocol 3: Maintenance of T. gondii tachyzoites using human foreskin fibroblastsSupport Protocol 4: Maintenance of T. gondii tachyzoites in miceSupport Protocol 5: Preparation of T. gondii lysate antigensSupport Protocol 6: Isolation of T. gondii tissue cysts from brain