CPHEN‐011: Comprehensive phenotyping of murine lung resident lymphocytes after recovery from pneumococcal pneumonia
| Autor: | Carolina Lyon De Ana, Kimberly A. Barker, Joseph P. Mizgerd, Anukul T. Shenoy, E.I. Arafa, I.M.C. Martin, Anna C. Belkina |
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| Rok vydání: | 2021 |
| Předmět: |
Histology
Lung medicine.diagnostic_test biology Cell Biology Pneumonia Pneumococcal CD8-Positive T-Lymphocytes medicine.disease Acquired immune system Phenotype Pathology and Forensic Medicine Flow cytometry Mice medicine.anatomical_structure Immune system Immunity Immunology Pneumococcal pneumonia medicine biology.protein Animals Lymphocytes Antibody Immunologic Memory |
| Zdroj: | Cytometry Part A. 101:892-902 |
| ISSN: | 1552-4930 1552-4922 |
| DOI: | 10.1002/cyto.a.24522 |
| Popis: | Recovery from pneumococcal (Spn) pneumonia induces development of tissue resident memory CD4+ TRM cells, BRM cells, and antibody secreting plasma cells in experienced lungs. These tissue resident lymphocytes confer protection against subsequent lethal challenge by serotype mismatched Spn (termed as heterotypic immunity). While traditional flow cytometry and gating strategies support premeditated identification of cells using a limited set of markers, discovery of novel tissue resident lymphocytes necessitates stable platforms that can handle larger sets of phenotypic markers and lends itself to unbiased clustering approaches. In this report, we leverage the power of full spectrum flow cytometry (FSFC) to develop a comprehensive panel of phenotypic markers that allows identification of multiple subsets of tissue resident lymphocytes in Spn-experienced murine lungs. Using Phenograph algorithm on this multidimensional data, we identify unforeseen heterogeneity in lung resident adaptive immune landscape which includes unexpected subsets of TRM and BRM cells. Further, using conventional gating strategy informed by our unsupervised clustering data, we confirm their presence exquisitely in Spn-experienced lungs as potentially relevant to heterotypic immunity and define CD73 as a highly expressed marker on TRM cells. Thus, our study emphasizes the utility of FSFC for confirmatory and discovery studies relating to tissue resident adaptive immunity. |
| Databáze: | OpenAIRE |
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