Supplementary Material
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Supplementary material ERJ-01469-2021.Supplement
Supplementary table S1 ERJ-01469-2021.Table
Supplementary figure S1. Gating strategy for mouse PBMCs and mouse lung cell flow cytometry analysis. Mouse PBMCs and dissociated lung cells were stained with a cocktail of fluorescently labelled monoclonal antibodies (mAbs) and analysed by flow cytometry. The leukocyte population was determined by gating CD45.2+ cells. CD45 is a general leukocyte cell surface marker. Leukocytes were subsequently stratified into specific immune cell subsets. a) PCs were identified by selecting CD19+ CD3− cells and then selecting CD38+ and CD138+ cells. b) B regulatory cells were identified by selecting CD5+ and then CD19+. ERJ-01469-2021.Figure_S1
Supplementary figure S2. Gating strategy for human white blood cell analysis using flow cytometry. Representative flow cytometry plots were shown for white blood cells from one control patient that was washed and stained with a cocktail of fluorescently labelled mAbs to distinguish T and B cell subsets. Lymphocytes were gated on FSC, and SSC properties and viable cells were gated and analysed for CD3+ T cells and CD19+ B cells. Mature B cells were identified as CD19+ CD20+ cells. These cells were then analysed for the presence of CD20+ CD27+ memory B cells (Bmem) and the Bmem cells were further analysed for the presence of CD20+ CD38+ plasmablasts (PBs). ERJ-01469-2021.Figure_S2
Supplementary figure S3. a) Low magnification image of a representative lung section from Blm + vehicle treated mice stained with H&E. The image shows areas of normal tissue and dense fibrosis, and multiple mononuclear cell aggregates are present within areas of fibrosis. Inset is enlarged in b) with a mononuclear cell aggregate (arrow). b) High magnification image of mononuclear cell aggregate contains CD19+CD138+ PCs (yellow). The isotype control shows no positive CD19 or CD138 staining. Scale bars 100 μm. ERJ-01469-2021.Figure_S3