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Monitoring Group 2 Innate Lymphoid Cell Biology in Models of Lung Inflammation
Innate lymphoid cells (ILCs) are a rare cell population subdivided into ILC1s, ILC2s, and ILC3s, based on transcription factor expression and cytokine production. In models of lung inflammation, the release of alarmins from the epithelium activates ILC2s and promotes the production of Th2-cytokines...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Bio-Protocol
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10366677/ https://www.ncbi.nlm.nih.gov/pubmed/37497449 http://dx.doi.org/10.21769/BioProtoc.4717 |
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author | Badrani, Jana H. Strohm, Allyssa N. Haung, Yung-An Doherty, Taylor A. |
author_facet | Badrani, Jana H. Strohm, Allyssa N. Haung, Yung-An Doherty, Taylor A. |
author_sort | Badrani, Jana H. |
collection | PubMed |
description | Innate lymphoid cells (ILCs) are a rare cell population subdivided into ILC1s, ILC2s, and ILC3s, based on transcription factor expression and cytokine production. In models of lung inflammation, the release of alarmins from the epithelium activates ILC2s and promotes the production of Th2-cytokines and the proliferation and migration of ILC2s within the lung. ILC2s are the innate counterpart to CD4(+) Th2s and, as such, express Gata-3 and produce IL-4, IL-5, and IL-13. Due to the low number of ILCs and the lack of specific surface markers, flow cytometry is the most reliable technique for the identification and characterization of ILCs. In this protocol, multicolor flow cytometry is utilized to identify Lineage- Thy1.2+ ILCs. Intracellular cytokine staining further identifies ILC2s within the lung. This protocol presents a reliable method for promoting ILC2-mediated lung inflammation and for monitoring ILC2 biology. Key features In this protocol, ILC2s are expanded via intranasal challenges withAlternaria alternata, a fungal allergen, or recombinant IL-33. Bronchoalveolar lavage (BAL) and lung are collected and processed into single-cell suspension for multicolor flow cytometric analysis, including intracellular staining of transcription factors and cytokines. During lung inflammation, the percentage of ILC2s and eosinophils increases. ILC2s express greater levels ofGata-3andKi-67and produce greater amounts of IL-5 and IL-13. Graphical overview [Image: see text] |
format | Online Article Text |
id | pubmed-10366677 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Bio-Protocol |
record_format | MEDLINE/PubMed |
spelling | pubmed-103666772023-07-26 Monitoring Group 2 Innate Lymphoid Cell Biology in Models of Lung Inflammation Badrani, Jana H. Strohm, Allyssa N. Haung, Yung-An Doherty, Taylor A. Bio Protoc Methods Article Innate lymphoid cells (ILCs) are a rare cell population subdivided into ILC1s, ILC2s, and ILC3s, based on transcription factor expression and cytokine production. In models of lung inflammation, the release of alarmins from the epithelium activates ILC2s and promotes the production of Th2-cytokines and the proliferation and migration of ILC2s within the lung. ILC2s are the innate counterpart to CD4(+) Th2s and, as such, express Gata-3 and produce IL-4, IL-5, and IL-13. Due to the low number of ILCs and the lack of specific surface markers, flow cytometry is the most reliable technique for the identification and characterization of ILCs. In this protocol, multicolor flow cytometry is utilized to identify Lineage- Thy1.2+ ILCs. Intracellular cytokine staining further identifies ILC2s within the lung. This protocol presents a reliable method for promoting ILC2-mediated lung inflammation and for monitoring ILC2 biology. Key features In this protocol, ILC2s are expanded via intranasal challenges withAlternaria alternata, a fungal allergen, or recombinant IL-33. Bronchoalveolar lavage (BAL) and lung are collected and processed into single-cell suspension for multicolor flow cytometric analysis, including intracellular staining of transcription factors and cytokines. During lung inflammation, the percentage of ILC2s and eosinophils increases. ILC2s express greater levels ofGata-3andKi-67and produce greater amounts of IL-5 and IL-13. Graphical overview [Image: see text] Bio-Protocol 2023-07-20 /pmc/articles/PMC10366677/ /pubmed/37497449 http://dx.doi.org/10.21769/BioProtoc.4717 Text en ©Copyright : © 2023 The Authors; This is an open access article under the CC BY license https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Methods Article Badrani, Jana H. Strohm, Allyssa N. Haung, Yung-An Doherty, Taylor A. Monitoring Group 2 Innate Lymphoid Cell Biology in Models of Lung Inflammation |
title | Monitoring Group 2 Innate Lymphoid Cell Biology in Models of Lung Inflammation |
title_full | Monitoring Group 2 Innate Lymphoid Cell Biology in Models of Lung Inflammation |
title_fullStr | Monitoring Group 2 Innate Lymphoid Cell Biology in Models of Lung Inflammation |
title_full_unstemmed | Monitoring Group 2 Innate Lymphoid Cell Biology in Models of Lung Inflammation |
title_short | Monitoring Group 2 Innate Lymphoid Cell Biology in Models of Lung Inflammation |
title_sort | monitoring group 2 innate lymphoid cell biology in models of lung inflammation |
topic | Methods Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10366677/ https://www.ncbi.nlm.nih.gov/pubmed/37497449 http://dx.doi.org/10.21769/BioProtoc.4717 |
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