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. 2011 Nov;219(5):611-21.
doi: 10.1111/j.1469-7580.2011.01415.x. Epub 2011 Jul 22.

Distribution, organization and innervation of gastric MALT in conventional piglet

Maurizio Mazzoni  1 , Paolo Bosi, Nadia De Sordi, Giovanna Lalatta-Costerbosa
Affiliations

Distribution, organization and innervation of gastric MALT in conventional piglet

Maurizio Mazzoni et al. J Anat. 2011 Nov.

Abstract

Mucosa-associated lymphoid tissue (MALT) is the initial inductive site for mucosal immunity. It is present in the different layers of the mucosal wall and consists of organized lymphoid tissue which may occur as isolated or aggregated lymphoid follicles (LFs) and interfollicular areas. It is present in many organs, including the pig stomach. Gastric MALT has been intensely studied in experimentally infected pigs but few data are available in healthy, non-gnotobiotic or germ-free animals. In the present study we described the gastric MALT in conventional piglets in the cardiac mucosa of the gastric diverticulum, in the pyloric mucosa, and in the sites of transition from cardiac to oxyntic and from cardiac to pyloric mucosa by means of histological and immunohistochemical stains. The majority of LFs were located in the cardiac mucosa and in the transition from the cardiac to the oxyntic mucosa. Here the LFs were mainly located in the submucosa and reached the mucosa; we called these submucosal lymphoid follicles (SLFs). In the pyloric mucosa and in the transition sites from the cardiac to the pyloric mucosa, LFs were located in the mucosa; we called these mucosal lymphoid follicles (MLFs). In SLFs, a compartmental organization of T and B lymphocytes was present; by contrast, in the MLFs, the T and B cells were intermingled, suggesting the possibility of different roles for the two types of follicles. In the epithelium overlying the lymphoid tissue, numerous T lymphocytes and some cells immunoreactive to cytokeratin-18 were observed. Following the application of the fluorescent tracer DiI into the SLFs of the diverticulum, enteric neurones located in the submucosal plexus were labelled, confirming the interplay between the immune and the enteric nervous system.

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Figures

Fig. 1
Fig. 1
The stomach of a piglet opened along the greater curvature, from the diverticulum to the pyloric sphincter to show the sites of sampling (1–8).
Fig. 2
Fig. 2
Transverse section of piglet gastric mucosa stained by Masson's trichrome. (A) A single ovoid submucosal lymphoid follicle (arrowhead) crossing the muscularis mucosae. (B) A single mucosal lymphoid follicle confined to the lamina propria without crossing the muscularis mucosae (arrowheads). Bars: (A) 800 μm; (B) 200 μm.
Fig. 3
Fig. 3
Serial transverse section of a submucosal (A–C) and a mucosal (D–F) lymphoid follicle stained with Masson's trichrome (A,D), CD3 (B,E) and CD79α (C,F). Note the compartmental organization of the lymphocytes in submucosal follicles; the T lymphocytes were confined to the mantle zone (B) and the B lymphocytes to the germinal centre (C). In the mucosal follicles, the T and B lymphocytes were intermingled (E,F) and a sub-epithelial dome region was evident (D, arrowhead). Bars: (A–C) 400 μm; (D–F) 200 μm.
Fig. 4
Fig. 4
Tangential serial sections of piglet gastric mucosa showing the epithelium forming a diverticulum which enters the lymphoid tissue. (E,F) Lymphoid follicles were aggregated to form PP-like structures. Bars: (A–F) 800 μm.
Fig. 5
Fig. 5
Transverse section of piglet gastric mucosa stained with anti-CD3 (A,B) and anti-cytokeratin 18 antibody (C–D). T lymphocytes were densely packed at the base of a submucosal lymphoid follicle (A) and infiltrated the epithelium overlying a submucosal lymphoid follicle (B). Columnar cytokeratin 18-IR cells either single (C) or grouped (D) were intermingled with epithelial cells. A network of fibrils filled the cytoplasm and surrounded the unlabelled nucleus (D, arrowed). Bars: (A,B) 200 μm; (C) 50 μm; (D) 30 μm.
Fig. 6
Fig. 6
Tangential sections of pig gastric mucosa at the level of the diverticulum. (A) Stereomicroscopy showing DiI-coated beads (arrow) applied to a follicle in fixed tissue after 8 months of incubation. (B,C,E) DiI crystals applied to the follicles; note that the tracer was homogeneously distributed inside the follicles. (B) Labelled neurones (arrowhead) and processes entering the labelled follicle. (C) Polyhedral-labelled neurone (arrowhead) more than 400 μm from a labelled follicle. (D) Higher magnification of the neurone shown in (C). (E) Elongated neurone (about 400 μm from the labelled follicle) along a thin nervous fascicle (arrowhead). (F) Higher magnification of the neurone shown in (E). (G,H) Two labelled neurones with an ovoid shape. Note that the neurones showed smoothly contoured soma and an eccentrically located nucleus (D,G,H,). Bars: (B,F) 100 μm; (C,E) 200 μm; (D,G,H) 30 μm.
Fig. 7
Fig. 7
Transverse sections of pig gastric mucosa at the level of the diverticulum after the insertion of DiI crystals in the lamina propria. (A) Note a thin plexus of labelled fibres between the adenomers. (B) A neurone (arrowhead) in the lamina propria projecting to the epithelium. (C) Labelled neurones (arrowhead) projecting into a labelled follicle. (D,E) Labelled fibres in unlabelled follicles. (F) Labelled fibres resting on the connective follicular capsule. (G) Labelled, ovoid neurone (arrowhead) near the labelled muscularis mucosae. (E,F) Merging with the green filtre cube to best show the lymphoid follicle structure. Bars: (A,E,F) 200 μm; (B,C,G) 100 μm; (D) 50 μm.
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