FIGURE LEGENDS
Figure 1. Flow cytometry and mass cytometry identify similar T cell subpopulations. (A) PBMCs and cells from control nasal tissues (Cont) and nasal polyps (NP) were stained for CD3+ T cell subsets using flow (left) or mass (right) cytometry. Ratios of CD4+ to CD8+ cell frequencies within the CD3+ T cell population are shown as means ± SEMs. Flow cytometry, n=5 PBMC, 2 Cont, and 12 NP; Mass cytometry, n=3 PBMC, 2 Cont, and 6 NP. (B) CD3+ cells were analyzed biaxially for CD4+ and CD8+ expression. Flow (left) and mass (right) cytometry data are depicted as stacked means of CD4-CD8- (double negative, DN), CD4+CD8+ (double positive, DP), CD4-CD8+ (CD8+) and CD4+CD8-(CD4+). (C) Proportions of CD69+cells within CD4+ T cells determined by flow (left) and mass (right) cytometry are shown as means ± SEMs. Circles represent individual participants. (D) Proportions of CD69+ cells within CD8+ T cells determined by mass cytometry are shown as means ± SEMs. Circles represent individual participants. (E and F) CD45+leukocytes from mass cytometry data were clustered based on expression of canonical markers (see Supplemental Table E3 and Supplemental Figure E2). Frequency of naïve CD4+ and CD8+ T cells (Panel E) and CD8+central memory T (TCM) cells (Panel F) within the CD3+ population are shown as box and whisker plots. n=3 PBMC, 2 Cont, and 6 NP. *, p<0.05; **, p<0.01 between groups indicated by horizontal lines.
Figure 2. Identification of unique T cell and lymphoid cell populations in nasal polyps by mass cytometry. (A) MDS map of CD45+ leukocytes in nasal polyps after unsupervised clustering based on expression of surface markers. (B) Frequency of naïve CD8+ T cell subpopulations within CD3+ T cells in nasal polyps (NP) and control nasal tissues (Cont) is presented. (C) Frequency of the CD161hiCXCR3loCD127loCD4+ TCM population within CD3+ T cells in nasal polyps (NP) and control nasal tissues (Cont) is presented. ND, not detected. (D) viSNE mapping revealed clusters of T cell populations in CD45+CD3+CD19-cells; representative nasal polyp (top) and control sinus tissue (bottom) are presented. Colors indicate marker expression levels as shown by the color key. Boxed cell population is unique to nasal polyps and minimally represented in control sinus tissues. (E) Frequency of a CD4+CD161hiCXCR3locell population (Panel D, boxed population) within CD45+CD3+CD19-cells is shown. Each circle represents an individual subject. (F) viSNE mapping of CD45+CD3-CD19-cells revealed a cluster of unique lymphoid cells (red circles) in nasal polyps but not in control sinus tissues; representative nasal polyp (top) and control sinus tissue (bottom) maps are presented. (G) Frequency of ILC2-like CD25+CRTH2+CD69+cells (Panel F, circled population) within the CD45+CD3-CD19-cell population is shown. Data in Panels B, C, E, and G are shown as means ± SEMs. n=6 NP and n=2 Cont. *, p<0.05; **, p<0.01 between groups indicated by horizontal lines.