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Parallel Propagation of Toxoplasma gondii In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model

Toxoplasma gondii is an obligate intracellular protozoan. In pregnant women, it can lead to severe birth defects or intrauterine death of the fetus. Most of what is currently know on cell biology of T. gondii comes from studies relying on the RH strain propagated in mice. According to the recommenda...

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Autores principales: Gargaté, Maria João, Vilares, Anabela, Ferreira, Idalina, Reis, Tânia, Martins, Susana, Mendonça, Joana, Borges, Vítor, Gomes, João Paulo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9502748/
https://www.ncbi.nlm.nih.gov/pubmed/36145470
http://dx.doi.org/10.3390/pathogens11091038
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author Gargaté, Maria João
Vilares, Anabela
Ferreira, Idalina
Reis, Tânia
Martins, Susana
Mendonça, Joana
Borges, Vítor
Gomes, João Paulo
author_facet Gargaté, Maria João
Vilares, Anabela
Ferreira, Idalina
Reis, Tânia
Martins, Susana
Mendonça, Joana
Borges, Vítor
Gomes, João Paulo
author_sort Gargaté, Maria João
collection PubMed
description Toxoplasma gondii is an obligate intracellular protozoan. In pregnant women, it can lead to severe birth defects or intrauterine death of the fetus. Most of what is currently know on cell biology of T. gondii comes from studies relying on the RH strain propagated in mice. According to the recommendations concerning the animal welfare, we assayed in vitro/in vivo procedures to replace, or at least reduce, the demanding animal model for strain propagation. We evaluated the genetic and phenotypic stability of the RH strain throughout its parallel continuous propagation in mice, in human foreskin fibroblasts (HFF) and in an alternate fashion of these two procedures. We also assessed the virulence impact on the RH strain after different periods of its long-term propagation strictly in cells. The RH strain completely lost its virulence after long-term passage in HFF. Nevertheless, we obtained a successful outcome with the alternate passaging of the parasite in HFF and in mice as this approach enabled T. gondii to maintain the evaluated phenotypic properties, mainly its virulence potential. Also, no genetic changes were observed in genes known to be highly polymorphic or involved in pathoadaptation. In conclusion, the alternate model seems to be a feasible method for T. gondii propagation and maintenance, strongly impacting the number of sacrificed mice.
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spelling pubmed-95027482022-09-24 Parallel Propagation of Toxoplasma gondii In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model Gargaté, Maria João Vilares, Anabela Ferreira, Idalina Reis, Tânia Martins, Susana Mendonça, Joana Borges, Vítor Gomes, João Paulo Pathogens Article Toxoplasma gondii is an obligate intracellular protozoan. In pregnant women, it can lead to severe birth defects or intrauterine death of the fetus. Most of what is currently know on cell biology of T. gondii comes from studies relying on the RH strain propagated in mice. According to the recommendations concerning the animal welfare, we assayed in vitro/in vivo procedures to replace, or at least reduce, the demanding animal model for strain propagation. We evaluated the genetic and phenotypic stability of the RH strain throughout its parallel continuous propagation in mice, in human foreskin fibroblasts (HFF) and in an alternate fashion of these two procedures. We also assessed the virulence impact on the RH strain after different periods of its long-term propagation strictly in cells. The RH strain completely lost its virulence after long-term passage in HFF. Nevertheless, we obtained a successful outcome with the alternate passaging of the parasite in HFF and in mice as this approach enabled T. gondii to maintain the evaluated phenotypic properties, mainly its virulence potential. Also, no genetic changes were observed in genes known to be highly polymorphic or involved in pathoadaptation. In conclusion, the alternate model seems to be a feasible method for T. gondii propagation and maintenance, strongly impacting the number of sacrificed mice. MDPI 2022-09-13 /pmc/articles/PMC9502748/ /pubmed/36145470 http://dx.doi.org/10.3390/pathogens11091038 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Gargaté, Maria João
Vilares, Anabela
Ferreira, Idalina
Reis, Tânia
Martins, Susana
Mendonça, Joana
Borges, Vítor
Gomes, João Paulo
Parallel Propagation of Toxoplasma gondii In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model
title Parallel Propagation of Toxoplasma gondii In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model
title_full Parallel Propagation of Toxoplasma gondii In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model
title_fullStr Parallel Propagation of Toxoplasma gondii In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model
title_full_unstemmed Parallel Propagation of Toxoplasma gondii In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model
title_short Parallel Propagation of Toxoplasma gondii In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model
title_sort parallel propagation of toxoplasma gondii in vivo, in vitro and in alternate model: towards less dependence on the mice model
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9502748/
https://www.ncbi.nlm.nih.gov/pubmed/36145470
http://dx.doi.org/10.3390/pathogens11091038
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