Supplementary Material
Please note: supplementary material is not edited by the Editorial Office, and is uploaded as it has been supplied by the author.
Supplementary material ERJ-01214-2017_Supplement
Figure S1. Pulmonary arterial isolation procedure. Pulmonary arteries(*) were identified under a stereomicroscope due to their close vicinity to airways(§). Arteries were exposed by gradual removal of lung tissue until ~1-2cm of artery was visible. The artery was then removed en-bloc including side branches and forwarded to cell isolation. Scale bar represents 1 cm. ERJ-01214-2017_Figure_S1
Figure S2. Lung morphology in donor and IPAH samples. Representative images of sections derived from explanted lung samples (n=4). Magnified areas containing pulmonary vessels are also shown. ERJ-01214-2017_Figure_S2
Figure S3. Correlation plots of cell populations measured in different flow cytometry panels. A) Total CD45+ cells, B) PMN: polymorphonuclear neutrophils, C) CD3+ T cells, D) Mac: macrophages, and E) Mast cells. P values and rho (r) estimates derived from Spearman's rank correlation are shown for each correlation. ERJ-01214-2017_Figure_S3
Figure S4. Correlation analysis of individual cell populations in matched lung samples. Analysed cell populations in the two separate pieces of lung were compared by Spearman's rank correlation. P values and rho (r) estimates derived from Spearman's rank correlation are shown. ERJ-01214-2017_Figure_S4
Figure S5. Abundance of cell populations excluded from global principal component analysis. Scatterplot and boxplot overlays of cell populations in IPAH and donor lungs as analysed by flow cytometry. NK, natural killer cells; NKT, natural killer T cells. Boxplots show median and interquartile range. ERJ-01214-2017_Figure_S5
Figure S6. Patient-to-patient variability in CD45+ cell populations. Heatmap representation presenting the patient-to-patient variability of the log odds ratio derived from the percentage CD45+ data for 18 individual cell populations. Blue colours indicate highest abundance, red lowest for a single cell type. PMN, polymorphonuclear neutrophils; Macs, Macrophages; Monos, monocytes; DC, dendritic cells; pDC, plasmacytoid DC; gd gamma delta. ERJ-01214-2017_Figure_S6
Figure S7. Euclidean clustering of individual IPAH and donor samples. Heatmap representation using Euclidean clustering of the log odds ratio derived from number of cells per mg tissue data for the 21 individual cell populations. Blue colours indicate highest abundance, red lowest for a single cell type. Monos, monocytes; DC dendritic cells, Macs, Macrophages; CTL, cytotoxic T cell; gd gamma delta; PMN, polymorphonuclear neutrophils. ERJ-01214-2017_Figure_S7
Figure S8. Abundance of remaining cell population in lung samples. Abundance of cell populations not included in Fig. 4 as determine by flow cytometric analysis of IPAH and donor lungs. DC, dendritic cells; PMN, polymorphonuclear neutrophils. Boxplots show median and interquartile range, statistical significance between groups was determined by the Wilcoxon rank sum test. ERJ-01214-2017_Figure_S8
Figure S9. Relative proportions of CD45+ cells in lung samples. The relative proportions of multiple cell populations in IPAH and donor lungs. DC, dendritic cells; pDC; plasmacytoid dendritic cells; PMN, polymorphonuclear neutrophils. Boxplots show median and interquartile range, statistical significance between groups was determined by the Wilcoxon rank sum test. ERJ-01214-2017_Figure_S9
Figure S10. Abundance of cell population in isolated pulmonary arteries. Remaining cell populations not included in Fig. 5 as measured by flow cytometric analysis in IPAH and donor samples. DC, dendritic cells; PMN, polymorphonuclear neutrophils. Boxplots show median and interquartile range, statistical significance between groups was determined by the Wilcoxon rank sum test. ERJ-01214-2017_Figure_S10