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Single cell RNA sequencing identifies unique inflammatory airspace macrophage subsets
Kara J. Mould, … , Max Seibold, William J. Janssen
Kara J. Mould, … , Max Seibold, William J. Janssen
Published February 5, 2019
Citation Information: JCI Insight. 2019;4(5):e126556. https://doi.org/10.1172/jci.insight.126556.
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Research Article Immunology Pulmonology

Single cell RNA sequencing identifies unique inflammatory airspace macrophage subsets

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Abstract

Macrophages are well recognized for their dual roles in orchestrating inflammatory responses and regulating tissue repair. In almost all acutely inflamed tissues, 2 main subclasses of macrophages coexist. These include embryonically derived resident tissue macrophages and BM-derived recruited macrophages. While it is clear that macrophage subsets categorized in this fashion display distinct transcriptional and functional profiles, whether all cells within these categories and in the same inflammatory microenvironment share similar functions or whether further specialization exists has not been determined. To investigate inflammatory macrophage heterogeneity on a more granular level, we induced acute lung inflammation in mice and performed single cell RNA sequencing of macrophages isolated from the airspaces during health, peak inflammation, and resolution of inflammation. In doing so, we confirm that cell origin is the major determinant of alveolar macrophage (AM) programing, and, to our knowledge, we describe 2 previously uncharacterized, transcriptionally distinct subdivisions of AMs based on proliferative capacity and inflammatory programing.

Authors

Kara J. Mould, Nathan D. Jackson, Peter M. Henson, Max Seibold, William J. Janssen

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Figure 1

Single cell transcriptional profiling identifies 5 discrete AM populations across homeostasis, acute inflammation, and resolving inflammation.

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Single cell transcriptional profiling identifies 5 discrete AM populatio...
Mice were treated with intratracheal LPS and macrophages were isolated from lavage at days 0 (homeostasis), 3 (peak neutrophil inflammation), and 6 (resolution of lung inflammation). (A) T-distributed stochastic neighbor embedding (tSNE) plot shows clustering of 902 cells based on gene expression. Point coordinates are based on tSNE dimensionality reduction of the top 6 principal components calculated from the 5,784 most informative genes. Cell color specifies assignment of cells to 1 of 5 clusters (c1–5) inferred using shared nearest neighbor clustering. (B) Normalized expression of macrophage markers overlaid on tSNE plot. (C) Time course information overlaid on tSNE plot. (D) Relative proportion of cells in each cluster versus time.

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