eprintid: 10108306
rev_number: 20
eprint_status: archive
userid: 608
dir: disk0/10/10/83/06
datestamp: 2020-08-24 11:39:05
lastmod: 2021-12-08 23:44:49
status_changed: 2020-08-24 11:39:05
type: article
metadata_visibility: show
creators_name: Ward, AI
creators_name: Lewis, MD
creators_name: Khan, AA
creators_name: McCann, CJ
creators_name: Francisco, AF
creators_name: Jayawardhana, S
creators_name: Taylor, MC
creators_name: Kelly, JM
title: In Vivo Analysis of Trypanosoma cruzi Persistence Foci at Single-Cell Resolution
ispublished: pub
divisions: UCL
divisions: B02
divisions: D13
divisions: G22
keywords: Chagas disease, Trypanosoma cruzi, chronic, chronic persistence, colon, murine imaging, persistence, skeletal muscle, skin
note: © 2020 Ward et al. This is an openaccess article distributed under the terms of the Creative Commons Attribution 4.0
International license (https://creativecommons.org/licenses/by/4.0/).
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.IMPORTANCETrypanosoma 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.
date: 2020-08
date_type: published
official_url: https://doi.org/10.1128/mBio.01242-20
oa_status: green
full_text_type: pub
pmcid: PMC7407085
language: eng
primo: open
primo_central: open_green
verified: verified_manual
elements_id: 1808945
doi: 10.1128/mBio.01242-20
pii: mBio.01242-20
lyricists_name: McCann, Conor
lyricists_id: CMCCA07
actors_name: McCann, Conor
actors_id: CMCCA07
actors_role: owner
full_text_status: public
publication: mBio
volume: 11
number: 4
article_number: e01242
event_location: United States
citation:        Ward, AI;    Lewis, MD;    Khan, AA;    McCann, CJ;    Francisco, AF;    Jayawardhana, S;    Taylor, MC;           Ward, AI;  Lewis, MD;  Khan, AA;  McCann, CJ;  Francisco, AF;  Jayawardhana, S;  Taylor, MC;  Kelly, JM;   - view fewer <#>    (2020)    In Vivo Analysis of Trypanosoma cruzi Persistence Foci at Single-Cell Resolution.                   mBio , 11  (4)    , Article e01242.  10.1128/mBio.01242-20 <https://doi.org/10.1128/mBio.01242-20>.       Green open access   
 
document_url: https://discovery.ucl.ac.uk/id/eprint/10108306/7/McCann_In%20Vivo%20Analysis%20of%20Trypanosoma%20cruzi%20Persistence%20Foci%20at%20Single-Cell%20Resolution_VoR.pdf