CD25⁺CD127⁺Foxp3^- Cells Represent a Major Subpopulation of CD8⁺ T Cells in the Eye Chambers of Normal Mice - Fig 2

<p><b>Expression/co-expression of CD25 and Foxp3 (A) and CD25 and CD127 (B) on CD8</b>^<b>+</b><b>cells in mouse peripheral blood and aqueous humor.</b> The results are expressed as a percentage of CD25⁺Foxp3⁺, CD25⁺Foxp3^-, CD25^-Foxp3⁺, CD25^-Foxp3^- (A), CD25⁺CD127⁺, CD25⁺CD127^-, CD25^-CD127⁺ and CD25^-CD127^- cells (B) within CD8⁺ T lymphocyte population. Results are the mean (± SD) of 3 independent experiments for each type of labeling. Peripheral blood samples were collected from individual mice [n = 15 (A); n = 15 (B)], whereas each aqueous humor sample consisted of cells pooled from the eyeballs of 5 (or sometimes more) mice [n = 15 samples (A); n = 15 samples (B)]. Examples of cytograms (dot plots) from different samples for different experiments (C). On the basis of expression/co-expression of CD25 and Foxp3 or CD25 and CD127, CD8⁺ T cells were subdivided into the following subsets: CD25⁺Foxp3⁺, CD25⁺Foxp3^-, CD25^-Foxp3⁺, CD25^-Foxp3^- (panels 1 and 2), CD25⁺CD127⁺, CD25⁺CD127^-, CD25^-CD127⁺ and CD25^-CD127^- cells (panels 3 and 4). Fluorescence minus one (FMO) staining was used to confirm the gating strategy used to identify CD25- (panels 1 and 3), Foxp3- (panel 1) and CD127-expressing cells (panel 3). Gated CD4⁺ T cells from peripheral blood served as a positive control for Foxp3 staining (panel 2). *P < 0.001.</p

The distribution of single- and double-positive CD4⁺ and CD8⁺ T cells in mouse peripheral blood and aqueous humor.

<p>The results are expressed as percentages of CD4⁺CD8^-, CD4^-CD8⁺ and CD4⁺CD8⁺ T cells within the total lymphocyte population (A) and as percentages of CD8^high and CD8^low cells among the CD8⁺ T cell subset (B). Results are the mean (± SD) of 6 independent experiments. Peripheral blood samples were collected from individual mice (n = 30), whereas each aqueous humor sample consisted of cells pooled from the eyeballs of 5 (or sometimes more) mice (n = 30 samples). Examples of cytograms (dot plots) from different samples for different experiments (C). As the first step, the lymphocyte population in peripheral blood was gated on the basis of forward and side scatter (FSC and SSC, respectively; panel 1 and 3). The location of this gate served as a point of reference to set the lymphocyte gate for aqueous humor samples. CD4⁺CD8^-, CD4^-CD8⁺ and CD4⁺CD8⁺ T cell subsets were defined according to the expression of CD4 and CD8 within the gated lymphocyte subpopulation (panels 2 and 4). Relative to the intensity of CD8 expression, the CD8⁺ T cell population was subdivided into CD8^high and CD8^low cell subsets (panels 2 and 4). Fluorescence minus one (FMO) staining was used to confirm the gating strategy used to identify CD8-expressing cells (panels 2 and 4). *P < 0.01, **P < 0.001.</p

Ultrastructure of mucous cells in the pyloric glands.

<p>Note the three types of granules: 1) irregular, filled with electron-lucent material and “speckled” with electron-dense spots (arrows); 2) round, bipartite (double arrows); and 3) granules with intermediate properties.</p

Ultrastructure of the surface epithelium cells in the basal (A), middle (B, C), and upper (D, E) regions of the corpus gastric pits and mucus structure (F).

<p>(A) Pit cells containing electron dense granules in their apical parts. Note the bundles of tonofilaments (arrow). (B) Cells with numerous electron-dense granules that form oval clusters in the supranuclear cytoplasm. (C) Cells with numerous granules that vary in electron density and aggregate with each other. (D) Degenerating cells with flattened nuclei and cytoplasm filled by the mucous mass. (E) Cells with destroyed apical regions. Note the mucus is partially released from the cell. (F) Mucus ultrastructure. Cell debris and fine fibrils are visible.</p

Ultrastructure of chief cells in the proper gastric gland of the corpus mucosa.

<p>(A) Bipolar organization of chief cells. Note the well-developed Golgi apparatus (<i>AG</i>). L: lumen of the gland. (B) Numerous secretory granules, varying in size and electron density, located in the supranuclear cytoplasm. (C) Well-developed rough endoplasmic reticulum in the basal region.</p

Gross morphological features of the European beaver stomach.

<p>(A) An external view of the stomach. Asterisk: the CGG on the lesser curvature; <i>E</i>: oesophagus; <i>D</i>: duodenum. (B) The mucosa after cutting the stomach along the greater curvature. Note differences in mucosal colour between the corpus - <i>C</i> and pylorus - <i>P</i>. Asterisk: the CGG. (C) The CGG viewed from the stomach lumen. Note the large crater-like (arrow heads) and small (arrows) orifices. <i>Z</i>: gastroesophageal junction (D) The stomach cut along the lesser curvature. Note the continuity between the muscular coat of the CGG and the muscularis of the esophagus. Asterisk: the CGG; <i>E</i>: oesophagus. (E) Section of the CGG showing its internal organization. Arrows: branched tubes that open on the luminal mucosal surface.</p

Ultrastructure of the neck of a proper gastric gland in the stomach corpus.

<p>Mucous neck cells - <i>M</i>, parietal cell - <i>P</i>, and differentiating chief cell - <i>C</i> surrounding the lumen of the gland - <i>L</i>. Note the bipartite secretory granules (arrows) and well-developed Golgi apparatus – <i>AG</i> present in the mucous neck cells as well as the numerous cisternae of rough endoplasmic reticulum (asterisk) and round, electron-dense secretory granules (double arrows) in the developing chief cell.</p

Ultrastructure of endocrine cells in the proper gastric gland of the corpus mucosa.

<p>(A) Endocrine cell with well-developed endoplasmic reticulum and polymorphic granules. (B) Endocrine cell with small, round, electron-dense granules and numerous filaments.</p

Cellular composition of the proper gastric glands.

<p>(A) Fundus mucosa. (B) Corpus mucosa at the greater curvature opposite to the GCC. (C) Mucosa surrounding the CGG orifices. (D) Corpus mucosa close to the pylorus. (E) The CGG. Cells were counted in ca. 500 000 µm² areas that contained longitudinal sections through the gastric glands, as described in “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094590#s2" target="_blank">Material and methods</a>”. Total number of cells forming the glands was taken as 100%. The presented data are means and standard deviations (n = 12).</p

Surface epithelium cell morphology and mucus structure.

<p>Note differences in the appearance of cells that line various parts of the pits as well as the presence of cell debris in the mucus.</p