Volume 153, Issue 1, Pages (January 2018)

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Volume 153, Issue 1, Pages 77-86 (January 2018) Triggering Receptor Expressed on Myeloid Cells-2 Expression Tracks With M2-Like Macrophage Activity and Disease Severity in COPD  Derek E. Byers, MD, PhD, Kangyun Wu, PhD, Geoffrey Dang-Vu, MD, Xiaohua Jin, MD, Eugene Agapov, PhD, Xiaofeng Zhang, MD, John T. Battaile, MD, Kenneth Schechtman, PhD, Roger Yusen, MD, MPH, Richard A. Pierce, PhD, Michael J. Holtzman, MD  CHEST  Volume 153, Issue 1, Pages 77-86 (January 2018) DOI: 10.1016/j.chest.2017.09.044 Copyright © 2017 American College of Chest Physicians Terms and Conditions

Figure 1 A-C, TREM2 messenger RNA (mRNA) expression is increased in COPD in cohort 1. A, Levels of TREM1 and TREM2 mRNA in lung tissues from patients with Global Initiative for Chronic Obstructive Lung Disease stage IV COPD and non-COPD. B, Corresponding analysis of TREM2 vs DAP12, CD68, and CD206 mRNA levels in lung tissues from patients with COPD and non-COPD. C, TREM1 and TREM2 mRNA expression normalized to GAPDH mRNA level in lung tissues from patients with COPD and non-COPD. Each dot represents the value for a single subject. Boxes demonstrate the median (horizontal line) and 25th to 75th percentiles, and whiskers represent the range. r values are from Spearman correlation, and P values are from Mann-Whitney U test. CD68 expression was not significantly different between control and COPD tissues. NS = not significant. CHEST 2018 153, 77-86DOI: (10.1016/j.chest.2017.09.044) Copyright © 2017 American College of Chest Physicians Terms and Conditions

Figure 2 A-C, TREM-2 protein expression is increased in COPD in cohort 1. A, Levels of TREM-1 and TREM-2 protein in lung tissues from patients with COPD and non-COPD measured by enzyme-linked immunosorbent assay. Each dot represents the value for a single subject. Boxes demonstrate the median (horizontal line) and 25th to 75th percentiles, and whiskers represent the range. P values are from Mann-Whitney U test. B, Immunostaining for TREM-2 (red) and TREM-1, CD68, and CCR2 (green) in lung tissues patients with Global Initiative for Chronic Obstructive Lung Disease stage IV COPD. Bars = 50 μm. C, Flow cytometry analysis of TREM-2 on CD14+ myeloid cells in lung tissues from patients with COPD and non-COPD. Gating strategy is noted in the first panel. Results from CD14-positive lung cells labeled with anti-TREM2 (gray) and isotype control cells (red) are representative of five patients with COPD and four patients with non-COPD examined overall, respectively. TREM = Triggering Receptor Expressed on Myeloid Cells. See Figure 1 legend for expansion of other abbreviation. CHEST 2018 153, 77-86DOI: (10.1016/j.chest.2017.09.044) Copyright © 2017 American College of Chest Physicians Terms and Conditions

Figure 3 A-F, TREM2 messenger (mRNA) expression is increased in COPD in cohort 2. A, Levels of TREM1 and TREM2 mRNA (normalized to GAPDH) in lung tissues from patients with Global Initiative for Chronic Obstructive Lung Disease (GOLD) stage IV COPD and never smokers. B, Levels of TREM1 and TREM2 mRNA normalized to CD68 mRNA level in lung tissues from patients with GOLD stage IV COPD and never smokers. D, Levels of TREM2/TREM1 mRNA ratio in patients with GOLD stage IV COPD and never smokers. E, Analysis of TREM2/TREM1 mRNA ratio in lung tissue from all subjects in cohort 2 using the receiver operating characteristic curve to compare values in patients with no COPD or GOLD stage I or II COPD vs GOLD stage III or IV COPD. F, Analysis of TREM2/TREM1 mRNA ratio vs FEV1 percent predicted or FEV1/FVC in all subjects from cohort 2. In (A-F), each dot represents the value for a single subject. Boxes demonstrate the median (horizontal line) and 25th to 75th percentiles, and whiskers represent the range. r values are from Spearman correlation, and P values are from Mann-Whitney U test. CD68 expression was not significantly different between never smokers and GOLD stage IV COPD in (B), and did not correlate with FEV1 percent predicted or FEV1/FVC in (F). AUC = area under the curve; NS = not significant. CHEST 2018 153, 77-86DOI: (10.1016/j.chest.2017.09.044) Copyright © 2017 American College of Chest Physicians Terms and Conditions