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.