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

Figure 7.

Figure 7

Interstitial CD4 T cells from the oral mucosa have a CD44hi phenotype. Anesthetized C57BL/6J mice were given 1.25 µg of FITC-conjugated anti-CD45 (CD45:FITC) mAb in 200 µl of PBS via retro-orbital intravenous injection. Mice were euthanized at 3 min. Single-cell suspensions obtained from blood were stained with vitality dye Zombie NIR followed by rat anti-mouse CD45:PE, B220, CD3, CD4, CD8, CD11b, Ly6G, and CD44 mAbs to identify CD4 T cell populations. Cells were analyzed by flow cytometry. (A) Dot plots from blood and oral mucosa samples gated on live Ly6G− CD11b− leukocytes (CD45:PE+) from a representative mouse are shown. In oral mucosa samples, CD4 T cells were identified as interstitial (CD4+ CD45:FITC−) denoted by the a gate and contour plot; or circulating (CD4+ CD45:FITC+) denoted by the b gate and contour plot. Circulating (CD4+ CD45:FITC+) CD4 T cells in blood are bimodal with high and low CD44 expression (panel c). Percentage of cells within a drawn gate is given. (B) Summary data of the% of antigen-experienced (CD44hi) CD4 T cells found in blood, or in the interstitial (CD45:FITC−) or circulating (CD45:FITC+) compartments of the oral mucosa. Data are from two experiments of at least six mice and is displayed as mean ± SEM. Student’s two-tailed t-test was used to compare means.