<italic toggle="yes">In Vivo</italic> Analysis of <named-content content-type="genus-species">Trypanosoma cruzi</named-content> Persistence Foci at Single-Cell Resolution

ABSTRACT Infections with Trypanosoma cruzi are usually lifelong despite generating a strong adaptive immune response. Identifying the sites of parasite persistence is therefore crucial to understanding how T. cruzi avoids immune-mediated destruction. However, this is a major technical challenge, bec...

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Autores principales:	Alexander I. Ward, Michael D. Lewis, Archie A. Khan, Conor J. McCann, Amanda F. Francisco, Shiromani Jayawardhana, Martin C. Taylor, John M. Kelly
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	Trypanosoma cruzi Chagas disease chronic persistence murine imaging colon skeletal muscle Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/74fc772af41a456994e952224e9e0ba2
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spelling	oai:doaj.org-article:74fc772af41a456994e952224e9e0ba22021-11-15T15:56:43Z<italic toggle="yes">In Vivo</italic> Analysis of <named-content content-type="genus-species">Trypanosoma cruzi</named-content> Persistence Foci at Single-Cell Resolution10.1128/mBio.01242-202150-7511https://doaj.org/article/74fc772af41a456994e952224e9e0ba22020-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01242-20https://doaj.org/toc/2150-7511ABSTRACT Infections with Trypanosoma cruzi are usually lifelong despite generating a strong adaptive immune response. Identifying the sites of parasite persistence is therefore crucial to understanding how T. cruzi avoids immune-mediated destruction. However, this is a major technical challenge, because the parasite burden during chronic infections is extremely low. Here, we describe an integrated approach involving comprehensive tissue processing, ex vivo imaging, and confocal microscopy, which allowed us to visualize infected host cells in murine tissue with exquisite sensitivity. Using bioluminescence-guided tissue sampling, with a detection level of <20 parasites, we showed that in the colon, smooth muscle myocytes in the circular muscle layer are the most common infected host cell type. Typically, during chronic infections, the entire colon of a mouse contains only a few hundred parasites, often concentrated in a small number of cells each containing >200 parasites, which we term mega-nests. In contrast, during the acute stage, when the total parasite burden is considerably higher and many cells are infected, nests containing >50 parasites are rarely found. In C3H/HeN mice, but not BALB/c mice, we identified skeletal muscle as a major site of persistence during the chronic stage, with most parasites being found in large mega-nests within the muscle fibers. Finally, we report that parasites are also frequently found in the skin during chronic murine infections, often in multiple infection foci. In addition to being a site of parasite persistence, this anatomical reservoir could play an important role in insect-mediated transmission and have implications for drug development. IMPORTANCE Trypanosoma cruzi causes Chagas disease, the most important parasitic infection in Latin America. Major pathologies include severe damage to the heart and digestive tract, although symptoms do not usually appear until decades after infection. Research has been hampered by the complex nature of the disease and technical difficulties in locating the extremely low number of parasites. Here, using highly sensitive imaging technology, we reveal the sites of parasite persistence during chronic-stage infections of experimental mice at single-cell resolution. We show that parasites are frequently located in smooth muscle cells in the circular muscle layer of the colon and that skeletal muscle cells and the skin can also be important reservoirs. This information provides a framework for investigating how the parasite is able to survive as a lifelong infection, despite a vigorous immune response. It also informs drug development strategies by identifying tissue sites that must be accessed to achieve a curative outcome.Alexander I. WardMichael D. LewisArchie A. KhanConor J. McCannAmanda F. FranciscoShiromani JayawardhanaMartin C. TaylorJohn M. KellyAmerican Society for MicrobiologyarticleTrypanosoma cruziChagas diseasechronic persistencemurine imagingcolonskeletal muscleMicrobiologyQR1-502ENmBio, Vol 11, Iss 4 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Trypanosoma cruzi Chagas disease chronic persistence murine imaging colon skeletal muscle Microbiology QR1-502
spellingShingle	Trypanosoma cruzi Chagas disease chronic persistence murine imaging colon skeletal muscle Microbiology QR1-502 Alexander I. Ward Michael D. Lewis Archie A. Khan Conor J. McCann Amanda F. Francisco Shiromani Jayawardhana Martin C. Taylor John M. Kelly <italic toggle="yes">In Vivo</italic> Analysis of <named-content content-type="genus-species">Trypanosoma cruzi</named-content> Persistence Foci at Single-Cell Resolution
description	ABSTRACT Infections with Trypanosoma cruzi are usually lifelong despite generating a strong adaptive immune response. Identifying the sites of parasite persistence is therefore crucial to understanding how T. cruzi avoids immune-mediated destruction. However, this is a major technical challenge, because the parasite burden during chronic infections is extremely low. Here, we describe an integrated approach involving comprehensive tissue processing, ex vivo imaging, and confocal microscopy, which allowed us to visualize infected host cells in murine tissue with exquisite sensitivity. Using bioluminescence-guided tissue sampling, with a detection level of <20 parasites, we showed that in the colon, smooth muscle myocytes in the circular muscle layer are the most common infected host cell type. Typically, during chronic infections, the entire colon of a mouse contains only a few hundred parasites, often concentrated in a small number of cells each containing >200 parasites, which we term mega-nests. In contrast, during the acute stage, when the total parasite burden is considerably higher and many cells are infected, nests containing >50 parasites are rarely found. In C3H/HeN mice, but not BALB/c mice, we identified skeletal muscle as a major site of persistence during the chronic stage, with most parasites being found in large mega-nests within the muscle fibers. Finally, we report that parasites are also frequently found in the skin during chronic murine infections, often in multiple infection foci. In addition to being a site of parasite persistence, this anatomical reservoir could play an important role in insect-mediated transmission and have implications for drug development. IMPORTANCE Trypanosoma cruzi causes Chagas disease, the most important parasitic infection in Latin America. Major pathologies include severe damage to the heart and digestive tract, although symptoms do not usually appear until decades after infection. Research has been hampered by the complex nature of the disease and technical difficulties in locating the extremely low number of parasites. Here, using highly sensitive imaging technology, we reveal the sites of parasite persistence during chronic-stage infections of experimental mice at single-cell resolution. We show that parasites are frequently located in smooth muscle cells in the circular muscle layer of the colon and that skeletal muscle cells and the skin can also be important reservoirs. This information provides a framework for investigating how the parasite is able to survive as a lifelong infection, despite a vigorous immune response. It also informs drug development strategies by identifying tissue sites that must be accessed to achieve a curative outcome.
format	article
author	Alexander I. Ward Michael D. Lewis Archie A. Khan Conor J. McCann Amanda F. Francisco Shiromani Jayawardhana Martin C. Taylor John M. Kelly
author_facet	Alexander I. Ward Michael D. Lewis Archie A. Khan Conor J. McCann Amanda F. Francisco Shiromani Jayawardhana Martin C. Taylor John M. Kelly
author_sort	Alexander I. Ward
title	<italic toggle="yes">In Vivo</italic> Analysis of <named-content content-type="genus-species">Trypanosoma cruzi</named-content> Persistence Foci at Single-Cell Resolution
title_short	<italic toggle="yes">In Vivo</italic> Analysis of <named-content content-type="genus-species">Trypanosoma cruzi</named-content> Persistence Foci at Single-Cell Resolution
title_full	<italic toggle="yes">In Vivo</italic> Analysis of <named-content content-type="genus-species">Trypanosoma cruzi</named-content> Persistence Foci at Single-Cell Resolution
title_fullStr	<italic toggle="yes">In Vivo</italic> Analysis of <named-content content-type="genus-species">Trypanosoma cruzi</named-content> Persistence Foci at Single-Cell Resolution
title_full_unstemmed	<italic toggle="yes">In Vivo</italic> Analysis of <named-content content-type="genus-species">Trypanosoma cruzi</named-content> Persistence Foci at Single-Cell Resolution
title_sort	<italic toggle="yes">in vivo</italic> analysis of <named-content content-type="genus-species">trypanosoma cruzi</named-content> persistence foci at single-cell resolution
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/74fc772af41a456994e952224e9e0ba2
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<italic toggle="yes">In Vivo</italic> Analysis of <named-content content-type="genus-species">Trypanosoma cruzi</named-content> Persistence Foci at Single-Cell Resolution

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