Linking Complement C3 and B Cells in Nasal Polyposis

Figure 5

Larger B cells contain more intracellular C3 in nasal polyps. Polyp () and inferior turbinate () tissue samples were shredded and digested to prepare single cell suspensions. After excluding doublets and dead cells, lymphocytes were pregated for CD45. CD3+, CD19+, and CD3-/CD19-/CD16-/CD56+ immune cells were evaluated for C3 expression. (a) Gating strategy of nasal polyp with corresponding inferior turbinate. Immune cell subsets were gated by CD19-APC vs. C3-AlexaFluor488® compared to C3-matched isotype control. CD3+ (orange) and CD3-/CD19-/CD16-/CD56+ (green) cells are shown together in the lower part of the plots (CD19-), whereas CD19+ cells are displayed in violet or pink (high C3 expression: C3++), and the cellular size was further displayed using FSC and SSC. (b) Histogram of nasal polyp and corresponding inferior turbinate presenting C3-AlexaFlour488® signal for antibody (dark grey) and isotype control (light grey) of CD45+/CD19+ B lymphocytes with C3++ population (red box). (c) The high C3 positivity (C3++) is presented as percentage of immune cell subsets for nasal polyps () compared to corresponding inferior turbinates () (CD19+ cells ). Means with SEM are shown. . (d) Intracellular C3 staining revealed a low C3-positivity for all cell types investigated in both tissues (NP , cIT ) shown as MFI of AlexaFluor488® (AF488, C3) for isotype and antibody whereby C3++ CD19+ cells were excluded. Means with SEM are shown. (e) Extracellular staining of C3 without any permeabilization presented by displaying MFI of C3 and isotype control for lymphocyte subsets in nasal polyps (). Means with SEM are shown.

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