Journal: Mucosal immunology
Article Title: Single-cell analysis of human nasal mucosal IgE antibody secreting cells reveals a newly minted phenotype
doi: 10.1016/j.mucimm.2023.02.008
Figure Lengend Snippet: B cell and plasma cell composition of NPs. A, primary population gating strategy for fluorescence-activated cell sorted CD19 + ASCs, CD19 − ASCs, and B cell subsets. One representative patient sample was selected for display (AFRS 1); B, B cell subset frequency of total CD19 + B cells in NPs; C, histograms of CD138 expression in naïve B cells from one donor (AFRS 1) and in ASCs from AFRS 1, 2, and 3 (left). CD19 + CD138 + cell frequency of total CD19 + ASCs (right); D, relative proportions of immunoglobulin isotype subclasses as a percentage of total VDJ sequences isolated ( n = 5907); E, stacked barplots showing the absolute number of ASCs per polyp (left) and the relative composition of each NP VDJ repertoire (right). AFRS = allergic fungal rhinosinusitis; ASC = antibody secreting cell; CD = cluster of differentiation; DN = double-negative B cells; Ig = immunoglobulin; NP = nasal polyp; SWM = switched memory; USM = unswitched memory; VDJ = immunoglobulin gene.
Article Snippet: An antibody cocktail was then added to the single-cell suspension, which included the following: normal mouse serum, FITC anti-human IgD (BD, clone IA6-2), Brilliant Violet 711 anti-human CD3 (Biolegend, clone OKT3), Brilliant Violet 711 anti-human CD14 (Biolegend, clone M5E2), PE/Cy7 anti-human CD19 (BD, clone SJ25C1), V450 anti-human CD38 (BD, clone HIT2), APC anti-human CD138 (Miltenyi Biotec, clone 44F9), APC-eFluor 780 anti-human CD27 (eBiosciences, clone O323), and LiveDead Aqua (Invitrogen).
Techniques: Fluorescence, Expressing, Isolation