Characterization of Immune Cells and Proinflammatory Mediators in the Pulmonary Environment
| Autor: | Wen Zheng, Breanne Y. Farris, Edwin C. K. Wan, Kelly L. Monaghan |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Chemokine General Chemical Engineering Article General Biochemistry Genetics and Molecular Biology Flow cytometry Proinflammatory cytokine Mice 03 medical and health sciences 0302 clinical medicine Immune system Cell Movement Animals Medicine Lung 030219 obstetrics & reproductive medicine General Immunology and Microbiology biology medicine.diagnostic_test business.industry Macrophages General Neuroscience Dendritic Cells Pneumonia Flow Cytometry Natural killer T cell medicine.disease 030104 developmental biology medicine.anatomical_structure Integrin alpha M Immunology biology.protein Chemokines Inflammation Mediators business |
| Zdroj: | J Vis Exp |
| ISSN: | 1940-087X |
| DOI: | 10.3791/61359 |
| Popis: | Immune cell expansion, activation, and trafficking to the lungs, which are controlled by the expression of multiple cytokines and chemokines, may be altered by severe brain injury. This is evidenced by the fact that pneumonia is a major cause of mortality in patients who have suffered from ischemic stroke. The goal of this protocol is to describe the use of multicolor flow cytometric analysis to identify 13 types of immune cells in the lungs of mice, including alveolar macrophages, interstitial macrophages, CD103+ or CD11b+ dendritic cells (DCs), plasmacytoid DCs, eosinophils, monocytes/monocyte-derived cells, neutrophils, lymphoid-derived T and B cells, NK cells, and NKT cells, following ischemic stroke induction by transient middle cerebral artery occlusion. Moreover, we describe the preparation of lung homogenates using a bead homogenization method, to determine the expression levels of 13 different cytokines or chemokines simultaneously by multiplex bead arrays coupled with flow cytometric analysis. This protocol can also be used to investigate the pulmonary immune response in other disease settings, such as infectious lung disease or allergic disease. |
| Databáze: | OpenAIRE |
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