Supplementary material

Supplementary material

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  • Supplementary materials and methods -
    Subjects
    Cell isolation from peripheral blood
    Lung cell isolation
    Flow cytometry analysis
    PBMC stimulation
    Immunocytochemistry
    IgG, IgA and IgM ELISA
    Antigen specific ELISPOT analysis
    Statistical analyses
  • Figure S1 - A schematic overview of the study design is depicted. PBMCs = peripheral blood mononuclear cells, ICC = immunocytochemistry, ELISPOT = Enzyme-linked immunosorbent spot, ELISA = Enzyme-linked immunosorbent assay.
  • Figure S2 - A) the gates for total B cells (CD20 positive) and memory B cells (CD20 and CD27 positive), B) the gate for class-switched memory B cells (CD27 positive and IgM negative) within the total B cell population, and C) the gates for IgG positive memory B cells, IgM positive memory B cells and memory B cells negative for IgG and IgM within the memory B cell population are shown.
  • Figure S3 - Percentages of total B cells, memory B cells, and class switched memory B cells in peripheral blood of COPD patients (closed symbols) and healthy individuals (open symbols) are shown. COPD patients had lower percentages of total B cells (COPD effect p=0.009) and memory B cells (COPD effect p= 0.029) than healthy individuals. Current smokers had higher percentages of memory B cells (smoke effect p=0.029) and class switched memory B cells (smoke effect p=0.001) than ex- and never smokers. The results of the Mann Whitney U tests are depicted in the figures. * indicates that p< 0.05.
  • Figure S4 - Percentages of IgE positive cells in peripheral blood of COPD patients (closed symbols) and healthy individuals (open symbols) are shown. COPD patients had lower percentages of IgE positive B cells than healthy individuals (COPD effect p=0.001).
  • Figure S5 - Total IgG (A) and IgM (B) antibody levels in supernatants from PBMCs derived from COPD patients (closed symbols) and healthy individuals (open symbols), which were cultured for 6 days without stimulation (left), with ECM + cytokine mix stimulation (middle) or cytokine mix stimulation (right) are shown. The results of the Mann Whitney U tests are depicted in the figures. * indicates that p< 0.05.
  • Figure S6 - Percentages of IgA and IgG plasma cells in cytospins from PBMCs derived from COPD patients (closed symbols) and healthy individuals (open symbols), which were cultured for 6 days with ECM + cytokine mix stimulation (left) or cytokine mix stimulation (right) are shown. The results of the Mann Whitney U tests are depicted in the figures. * indicates that p< 0.05.