Optimized Protocol for Characterization of Mouse Gut Innate Lymphoid Cells
| العنوان: | Optimized Protocol for Characterization of Mouse Gut Innate Lymphoid Cells |
|---|---|
| المؤلفون: | Maria José Gomez-Sánchez, Ana Valle-Noguera, Aranzazu Cruz-Adalia, Mathilde J. H. Girard-Madoux |
| المساهمون: | Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), European Commission |
| المصدر: | Frontiers in Immunology, Vol 11 (2020) Frontiers in Immunology Digital.CSIC. Repositorio Institucional del CSIC instname |
| بيانات النشر: | Frontiers Media S.A., 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0301 basic medicine, lcsh:Immunologic diseases. Allergy, Innate lymphoid cell type 3, Immunology, innate lymphoid cells, Innate lymphoid cells, Inflammation, Cell Separation, Biology, Lymphocyte Activation, Flow cytometry, Mice, 03 medical and health sciences, 0302 clinical medicine, Intestine, Small, Methods, medicine, Extracellular, Animals, Immunology and Allergy, Lymphocytes, innate lymphoid cell type 3, Intestinal Mucosa, Tissue homeostasis, ILC3 cytokines, Lamina propria, Innate immune system, medicine.diagnostic_test, colon, flow cytometry, Innate lymphoid cell, Lamina propria cells [Colon], lamina propria cells, Colon: Lamina propria cells, Gut immunology, Small intestine, Immunity, Innate, Cell biology, Mice, Inbred C57BL, 030104 developmental biology, medicine.anatomical_structure, Cytokines, medicine.symptom, lcsh:RC581-607, small intestine, Function (biology), 030215 immunology |
| الوصف: | © 2020 Valle-Noguera, Gómez-Sánchez, Girard-Madoux and Cruz-Adalia. Since their discovery, innate lymphoid cells (ILCs) have gradually been gaining greater relevance in the field of immunology due to their multiple functions in the innate immune response. They can mainly be found in mucosal and barrier organs like skin, gut, and lungs, and have been classified into five main types (NKs, ILC1s, ILC2s, ILC3s, and Lti cells) according to their function and development. They all play major roles in functions such as tissue homeostasis, early pathogen defense, regulation of inflammation, or tissue remodeling. ILCs are mostly tissue-resident cells tightly bound to the tissue structure, a fact that requires long and complex protocols that do not always provide sufficient yield for analysis. This suggests the need for optimized approaches aimed at ensuring that enriched and viable ILC samples are obtained, in order to furnish quality results. Herein a detailed protocol is established for obtaining a single-cell suspension highly enriched in lymphoid cells from mouse gut in order to identify the different subsets of ILCs by means of flow cytometry. The cell marker panel and flow cytometry gating strategies for identification and quantification of all the different ILC populations are provided for simultaneous analysis. Moreover, the protocol described includes a procedure for studying the different cytokines produced by ILC3s involved in maintaining the integrity of the gut barrier and defending against extracellular pathogens. As a result, herein an efficient method is presented for studying mouse ILCs within the lamina propria of the small intestine and colon; this can constitute a useful tool for future investigations in the field. The present research was supported by grant Nº RTI2018-093647-B-I00 to AC-A from Ministerio de Ciencia, Innovación e Universidades (MCIU), Agenda Estatal de Investigación (AEI), and Fondo Europeo de Desarrollo Regional (FEDER). AV-N is a recipient of an FPI fellowship (PRE2019-090341) from the Spanish Ministry of Science, Innovation, and Universities. |
| اللغة: | English |
| تدمد: | 1664-3224 |
| DOI: | 10.3389/fimmu.2020.563414/full |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9d73f41f0ffdf0be7c439c9a704f9558 https://www.frontiersin.org/articles/10.3389/fimmu.2020.563414/full |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....9d73f41f0ffdf0be7c439c9a704f9558 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 16643224 |
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| DOI: | 10.3389/fimmu.2020.563414/full |