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Flow Cytometric Analysis of Myeloid Cells in Human Blood, Bronchoalveolar Lavage, and Lung Tissues.

Publication ,  Journal Article
Yu, Y-RA; Hotten, DF; Malakhau, Y; Volker, E; Ghio, AJ; Noble, PW; Kraft, M; Hollingsworth, JW; Gunn, MD; Tighe, RM
Published in: Am J Respir Cell Mol Biol
January 2016

Clear identification of specific cell populations by flow cytometry is important to understand functional roles. A well-defined flow cytometry panel for myeloid cells in human bronchoalveolar lavage (BAL) and lung tissue is currently lacking. The objective of this study was to develop a flow cytometry-based panel for human BAL and lung tissue. We obtained and performed flow cytometry/sorting on human BAL cells and lung tissue. Confocal images were obtained from lung tissue using antibodies for cluster of differentiation (CD)206, CD169, and E cadherin. We defined a multicolor flow panel for human BAL and lung tissue that identifies major leukocyte populations. These include macrophage (CD206(+)) subsets and other CD206(-) leukocytes. The CD206(-) cells include: (1) three monocyte (CD14(+)) subsets, (2) CD11c(+) dendritic cells (CD14(-), CD11c(+), HLA-DR(+)), (3) plasmacytoid dendritic cells (CD14(-), CD11c(-), HLA-DR(+), CD123(+)), and (4) other granulocytes (neutrophils, mast cells, eosinophils, and basophils). Using this panel on human lung tissue, we defined two populations of pulmonary macrophages: CD169(+) and CD169(-) macrophages. In lung tissue, CD169(-) macrophages were a prominent cell type. Using confocal microscopy, CD169(+) macrophages were located in the alveolar space/airway, defining them as alveolar macrophages. In contrast, CD169(-) macrophages were associated with airway/alveolar epithelium, consistent with interstitial-associated macrophages. We defined a flow cytometry panel in human BAL and lung tissue that allows identification of multiple immune cell types and delineates alveolar from interstitial-associated macrophages. This study has important implications for defining myeloid cells in human lung samples.

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Published In

Am J Respir Cell Mol Biol

DOI

EISSN

1535-4989

Publication Date

January 2016

Volume

54

Issue

1

Start / End Page

13 / 24

Location

United States

Related Subject Headings

  • Young Adult
  • Sialic Acid Binding Ig-like Lectin 1
  • Respiratory System
  • Myeloid Cells
  • Microscopy, Confocal
  • Male
  • Macrophages, Alveolar
  • Lung
  • Immunophenotyping
  • Humans
 

Citation

APA
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ICMJE
MLA
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Yu, Y.-R., Hotten, D. F., Malakhau, Y., Volker, E., Ghio, A. J., Noble, P. W., … Tighe, R. M. (2016). Flow Cytometric Analysis of Myeloid Cells in Human Blood, Bronchoalveolar Lavage, and Lung Tissues. Am J Respir Cell Mol Biol, 54(1), 13–24. https://doi.org/10.1165/rcmb.2015-0146OC
Yu, Yen-Rei A., Danielle F. Hotten, Yuryi Malakhau, Ellen Volker, Andrew J. Ghio, Paul W. Noble, Monica Kraft, John W. Hollingsworth, Michael D. Gunn, and Robert M. Tighe. “Flow Cytometric Analysis of Myeloid Cells in Human Blood, Bronchoalveolar Lavage, and Lung Tissues.Am J Respir Cell Mol Biol 54, no. 1 (January 2016): 13–24. https://doi.org/10.1165/rcmb.2015-0146OC.
Yu Y-RA, Hotten DF, Malakhau Y, Volker E, Ghio AJ, Noble PW, et al. Flow Cytometric Analysis of Myeloid Cells in Human Blood, Bronchoalveolar Lavage, and Lung Tissues. Am J Respir Cell Mol Biol. 2016 Jan;54(1):13–24.
Yu, Yen-Rei A., et al. “Flow Cytometric Analysis of Myeloid Cells in Human Blood, Bronchoalveolar Lavage, and Lung Tissues.Am J Respir Cell Mol Biol, vol. 54, no. 1, Jan. 2016, pp. 13–24. Pubmed, doi:10.1165/rcmb.2015-0146OC.
Yu Y-RA, Hotten DF, Malakhau Y, Volker E, Ghio AJ, Noble PW, Kraft M, Hollingsworth JW, Gunn MD, Tighe RM. Flow Cytometric Analysis of Myeloid Cells in Human Blood, Bronchoalveolar Lavage, and Lung Tissues. Am J Respir Cell Mol Biol. 2016 Jan;54(1):13–24.

Published In

Am J Respir Cell Mol Biol

DOI

EISSN

1535-4989

Publication Date

January 2016

Volume

54

Issue

1

Start / End Page

13 / 24

Location

United States

Related Subject Headings

  • Young Adult
  • Sialic Acid Binding Ig-like Lectin 1
  • Respiratory System
  • Myeloid Cells
  • Microscopy, Confocal
  • Male
  • Macrophages, Alveolar
  • Lung
  • Immunophenotyping
  • Humans