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Precision cut lung slices: a novel versatile tool to examine host–pathogen interaction in the chicken lung

The avian respiratory tract is a common entry route for many pathogens and an important delivery route for vaccination in the poultry industry. Immune responses in the avian lung have mostly been studied in vivo due to the lack of robust, relevant in vitro and ex vivo models mimicking the microenvir...

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Autores principales: Bryson, Karen Jane, Garrido, Damien, Esposito, Marco, McLachlan, Gerry, Digard, Paul, Schouler, Catherine, Guabiraba, Rodrigo, Trapp, Sascha, Vervelde, Lonneke
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6954617/
https://www.ncbi.nlm.nih.gov/pubmed/31924278
http://dx.doi.org/10.1186/s13567-019-0733-0
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author Bryson, Karen Jane
Garrido, Damien
Esposito, Marco
McLachlan, Gerry
Digard, Paul
Schouler, Catherine
Guabiraba, Rodrigo
Trapp, Sascha
Vervelde, Lonneke
author_facet Bryson, Karen Jane
Garrido, Damien
Esposito, Marco
McLachlan, Gerry
Digard, Paul
Schouler, Catherine
Guabiraba, Rodrigo
Trapp, Sascha
Vervelde, Lonneke
author_sort Bryson, Karen Jane
collection PubMed
description The avian respiratory tract is a common entry route for many pathogens and an important delivery route for vaccination in the poultry industry. Immune responses in the avian lung have mostly been studied in vivo due to the lack of robust, relevant in vitro and ex vivo models mimicking the microenvironment. Precision-cut lung slices (PCLS) have the major advantages of maintaining the 3-dimensional architecture of the lung and includes heterogeneous cell populations. PCLS have been obtained from a number of mammalian species and from chicken embryos. However, as the embryonic lung is physiologically undifferentiated and immunologically immature, it is less suitable to examine complex host–pathogen interactions including antimicrobial responses. Here we prepared PCLS from immunologically mature chicken lungs, tested different culture conditions, and found that serum supplementation has a detrimental effect on the quality of PCLS. Viable cells in PCLS remained present for ≥ 40 days, as determined by viability assays and sustained motility of fluorescent mononuclear phagocytic cells. The PCLS were responsive to lipopolysaccharide stimulation, which induced the release of nitric oxide, IL-1β, type I interferons and IL-10. Mononuclear phagocytes within the tissue maintained phagocytic activity, with live cell imaging capturing interactions with latex beads and an avian pathogenic Escherichia coli strain. Finally, the PCLS were also shown to be permissive to infection with low pathogenic avian influenza viruses. Taken together, immunologically mature chicken PCLS provide a suitable model to simulate live organ responsiveness and cell dynamics, which can be readily exploited to examine host–pathogen interactions and inflammatory responses.
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spelling pubmed-69546172020-01-14 Precision cut lung slices: a novel versatile tool to examine host–pathogen interaction in the chicken lung Bryson, Karen Jane Garrido, Damien Esposito, Marco McLachlan, Gerry Digard, Paul Schouler, Catherine Guabiraba, Rodrigo Trapp, Sascha Vervelde, Lonneke Vet Res Research Article The avian respiratory tract is a common entry route for many pathogens and an important delivery route for vaccination in the poultry industry. Immune responses in the avian lung have mostly been studied in vivo due to the lack of robust, relevant in vitro and ex vivo models mimicking the microenvironment. Precision-cut lung slices (PCLS) have the major advantages of maintaining the 3-dimensional architecture of the lung and includes heterogeneous cell populations. PCLS have been obtained from a number of mammalian species and from chicken embryos. However, as the embryonic lung is physiologically undifferentiated and immunologically immature, it is less suitable to examine complex host–pathogen interactions including antimicrobial responses. Here we prepared PCLS from immunologically mature chicken lungs, tested different culture conditions, and found that serum supplementation has a detrimental effect on the quality of PCLS. Viable cells in PCLS remained present for ≥ 40 days, as determined by viability assays and sustained motility of fluorescent mononuclear phagocytic cells. The PCLS were responsive to lipopolysaccharide stimulation, which induced the release of nitric oxide, IL-1β, type I interferons and IL-10. Mononuclear phagocytes within the tissue maintained phagocytic activity, with live cell imaging capturing interactions with latex beads and an avian pathogenic Escherichia coli strain. Finally, the PCLS were also shown to be permissive to infection with low pathogenic avian influenza viruses. Taken together, immunologically mature chicken PCLS provide a suitable model to simulate live organ responsiveness and cell dynamics, which can be readily exploited to examine host–pathogen interactions and inflammatory responses. BioMed Central 2020-01-10 2020 /pmc/articles/PMC6954617/ /pubmed/31924278 http://dx.doi.org/10.1186/s13567-019-0733-0 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Bryson, Karen Jane
Garrido, Damien
Esposito, Marco
McLachlan, Gerry
Digard, Paul
Schouler, Catherine
Guabiraba, Rodrigo
Trapp, Sascha
Vervelde, Lonneke
Precision cut lung slices: a novel versatile tool to examine host–pathogen interaction in the chicken lung
title Precision cut lung slices: a novel versatile tool to examine host–pathogen interaction in the chicken lung
title_full Precision cut lung slices: a novel versatile tool to examine host–pathogen interaction in the chicken lung
title_fullStr Precision cut lung slices: a novel versatile tool to examine host–pathogen interaction in the chicken lung
title_full_unstemmed Precision cut lung slices: a novel versatile tool to examine host–pathogen interaction in the chicken lung
title_short Precision cut lung slices: a novel versatile tool to examine host–pathogen interaction in the chicken lung
title_sort precision cut lung slices: a novel versatile tool to examine host–pathogen interaction in the chicken lung
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6954617/
https://www.ncbi.nlm.nih.gov/pubmed/31924278
http://dx.doi.org/10.1186/s13567-019-0733-0
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