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, Vienna, Austria) and anti-human/rat IgG2b K isotype control-allophycocyanin-conjugated (eBioscience, Inc.) monoclonal immunoglobulins were used. Staining with fluorescein isothiocyanate-conjugated mouse anti-human CD14 (Beckman Coulter Company, Fullerton, CA, USA; Immunotech, Marseille, France) was performed to detect monocyte/macrophages and granulocytes, respectively. Cells were analysed on a NAVIOS (Beckman Coulter, Inc.) cytometer using CXP software (Beckman Coulter, Inc.). TREM-1 and TREM-2 expressions were measured as the total number and as Cell Cycle inhibitor the percentage of TREM-1 and TREM-2 positive cells out of all CD14+ cells and mean fluorescence intensity (MFI) of 10?000 cells in BALF subtracted from isotype control. The corresponding tests Thymidine kinase were called total number, percentage and MFI, respectively. The total number of TREM-1 and TREM-2 positive cells was calculated from the absolute count of BAL cells evaluated by flow cytometer (absolute count of cells per millilitre was obtained from filtered but not concentrated or centrifuged BAL by Flow-Count Fluor spheres (Beckman Coulter, Inc.). The one-sample Kolmogorov�CSmirnov test was used to determine whether the investigated population followed a normal distribution. Either non-parametric Mann�CWhitney U-test or parametric, non-paired t-test with Welch correction was used to determine differences and statistical significance. Fisher's exact test was used to determine specificity and sensitivity. We defined the cut-off for total number as more than 20 positive cells per microlitre (>20/��l), percentage as more than 20% of positive cells (>20% positive cells) and MFI superior to 4 (MFI;?>?4) for TREM-1 or TREM-2 positive cells. A value higher than the cut-off was a positive result and lower than the cut-off was negative. We found a significant increase in all tested variables: total number, percentage, and MFI of both TREM-1 and TREM-2 positive cells in PS patients compared with the control group. Characteristics of these groups are summarized in Table?1. Total number, percentage and MFI of TREM-1 positive cells were higher in PS patients (total number: median: 23.81 Quizartinib cells/��l, range: 6.89�C150.63; percentage: median: 41.30%, range: 13.30�C67.90; MFI: median: 5.43, range: 1.38�C12.50) in comparison with other ILD patients (total number: median: 13.50 cells/��l, range: 1.05�C123.95, P?=?0.0039; percentage: median: 15.70%, range: 1.20�C64.70, P?=?0.0002; MFI: median: 1.96, range: 0.48�C8.26, P?=?0.0005) (Figs?1-4). Total number, percentage and MFI of TREM-2 positive cells were higher in PS patients (total number: median: 32.81 cells/��l, range: 9.42�C81.25; percentage: median: 34%, range: 11.30�C82.30; MFI: median: 6.85, range: 1.92�C62.68) in comparison with other ILD patients (total number: median: 7.76 cells/��l, range: 0.28�C53.57, P?