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. 2017 Oct 30;8:1398. doi: 10.3389/fimmu.2017.01398

Figure 9.

Figure 9

Unaccounted circulating leukocytes skew frequency of granulocytes and B cells in oral mucosa samples. Anesthetized C57BL/6J or BALB/cJ mice were given anti-CD45:FITC mAb as described in Figure 7 and euthanized at 3 min. Oral mucosa were pooled from two mice and processed as single-cell suspensions. Cells were stained with vitality dye Zombie NIR followed by rat anti-mouse CD45:PE, CD3, CD8, CD4, B220, and Gr1 mAbs to identify live leukocyte cell subsets by flow cytometry. (A) Gating strategy used to identify live interstitial (Zombie NIRlow, CD45:PE+, CD45:FITC−) and circulating (Zombie NIRlow, CD45:PE−, CD45:FITC+) granulocytes (Gr1+, B220−, CD3−), B cells (B220+, Gr1−, CD3−), CD4 T cells (CD3+, CD4+, B220−), and CD8 T cells (CD3+, CD8+, B220−). Percentage of cells within a drawn gate is given. (B) Frequency of granulocytes, B cells, CD4 T cells, and CD8 T cells found in oral mucosa samples as a percentage of total immune cell populations (both circulating and interstitial) or interstitial immune cell populations alone. Data are displayed as mean ± SEM. Student’s two-tailed t-test was used to compare means (*p < 0.05, n = 4).