Induction of follicular gastritis following postthymectomy autoimmune gastritis in Helicobacter pylori-infected BALB/c mice

Abstract

Helicobacter pylori is the major causative agent of chronic antral gastritis and is thought to be involved in the pathogenesis of mucosa-associated lymphoid tissue lymphoma (MALToma) developing in the human stomach. The aim of this study was to clarify whether corporal autoimmune gastritis (AIG), which is known to decrease acidity due to destruction of parietal cells, predisposes mice to H. pylori infection, thereby leading to MALToma-like pathology. BALB/c mice in which AIG had been induced by thymectomy 3 days after birth (AIG mice) were used. The AIG mice were orally administered mouse-adapted H. pylori at the age of 6 weeks and were examined histologically and serologically after 2 to 12 months. The results were compared with those obtained from uninfected AIG mice and infected normal mice. Germinal centers were induced in the corpus in 57% of the H. pylori-infected AIG mice, which elicited anti-H. pylori antibody responses in association with upregulation of interleukin-4 (IL-4) mRNA. In these mice, parietal cells remained in the corpus mucosa. These findings were in contrast to those with the uninfected AIG mice: fundic gland atrophy due to disappearance of parietal cells associated with upregulation of gamma interferon, but not IL-4, mRNA and no germinal center formation in the corpus. These observations suggest that AIG alters the infectivity of H. pylori, leading to MALToma-like follicular gastritis, at an early stage after H. pylori infection.

FIG. 1

Sections of gastric mucosae of d3-Tx (a to f and h to j) and normal (g) BALB/c mice given saline or H. pylori orally at the age of 6 weeks. (a) Four months after saline administration; (b, c, d, f, g, h, i, and j) 4 months after H. pylori administration; (e) 12 months after H. pylori administration. (a) The body mucosa, showing the infiltration of mononuclear cells and gland atrophy characterized by loss of parietal and chief cells and replacement with proliferating epithelial cells. Magnification, ×125. (b) Germinal center formation in the body mucosa and persistence of parietal cells (arrowheads). Magnification, ×50. (c) Germinal center and LEL (arrows). Magnification, ×250. (d) Massive proliferation of plasma cells in the body mucosa. Magnification, ×250. (e) Mucin-producing cells and cells which have eosinophilic cytoplasm in the corpus. Magnification, ×125. (f) Colonization of the body pits by H. pylori. Magnification, ×500. (g) Low lymphocyte infiltration in the gastric mucosa in a normal BALB/c mouse after H. pylori infection. Magnification, ×125. (h, i, and j) Massive B220⁺ cells, scattered CD4⁺ cells, and no CD8⁺ cells, respectively, in the lymph follicle of a d3-Tx mouse 4 months after H. pylori infection. Magnification, ×250. (a to e and g) Hematoxylin-eosin staining; (f) May-Grunwald Giemsa staining; (h to j) immunohistochemical staining.

FIG. 1

Sections of gastric mucosae of d3-Tx (a to f and h to j) and normal (g) BALB/c mice given saline or H. pylori orally at the age of 6 weeks. (a) Four months after saline administration; (b, c, d, f, g, h, i, and j) 4 months after H. pylori administration; (e) 12 months after H. pylori administration. (a) The body mucosa, showing the infiltration of mononuclear cells and gland atrophy characterized by loss of parietal and chief cells and replacement with proliferating epithelial cells. Magnification, ×125. (b) Germinal center formation in the body mucosa and persistence of parietal cells (arrowheads). Magnification, ×50. (c) Germinal center and LEL (arrows). Magnification, ×250. (d) Massive proliferation of plasma cells in the body mucosa. Magnification, ×250. (e) Mucin-producing cells and cells which have eosinophilic cytoplasm in the corpus. Magnification, ×125. (f) Colonization of the body pits by H. pylori. Magnification, ×500. (g) Low lymphocyte infiltration in the gastric mucosa in a normal BALB/c mouse after H. pylori infection. Magnification, ×125. (h, i, and j) Massive B220⁺ cells, scattered CD4⁺ cells, and no CD8⁺ cells, respectively, in the lymph follicle of a d3-Tx mouse 4 months after H. pylori infection. Magnification, ×250. (a to e and g) Hematoxylin-eosin staining; (f) May-Grunwald Giemsa staining; (h to j) immunohistochemical staining.

FIG. 1

Sections of gastric mucosae of d3-Tx (a to f and h to j) and normal (g) BALB/c mice given saline or H. pylori orally at the age of 6 weeks. (a) Four months after saline administration; (b, c, d, f, g, h, i, and j) 4 months after H. pylori administration; (e) 12 months after H. pylori administration. (a) The body mucosa, showing the infiltration of mononuclear cells and gland atrophy characterized by loss of parietal and chief cells and replacement with proliferating epithelial cells. Magnification, ×125. (b) Germinal center formation in the body mucosa and persistence of parietal cells (arrowheads). Magnification, ×50. (c) Germinal center and LEL (arrows). Magnification, ×250. (d) Massive proliferation of plasma cells in the body mucosa. Magnification, ×250. (e) Mucin-producing cells and cells which have eosinophilic cytoplasm in the corpus. Magnification, ×125. (f) Colonization of the body pits by H. pylori. Magnification, ×500. (g) Low lymphocyte infiltration in the gastric mucosa in a normal BALB/c mouse after H. pylori infection. Magnification, ×125. (h, i, and j) Massive B220⁺ cells, scattered CD4⁺ cells, and no CD8⁺ cells, respectively, in the lymph follicle of a d3-Tx mouse 4 months after H. pylori infection. Magnification, ×250. (a to e and g) Hematoxylin-eosin staining; (f) May-Grunwald Giemsa staining; (h to j) immunohistochemical staining.

FIG. 1

Sections of gastric mucosae of d3-Tx (a to f and h to j) and normal (g) BALB/c mice given saline or H. pylori orally at the age of 6 weeks. (a) Four months after saline administration; (b, c, d, f, g, h, i, and j) 4 months after H. pylori administration; (e) 12 months after H. pylori administration. (a) The body mucosa, showing the infiltration of mononuclear cells and gland atrophy characterized by loss of parietal and chief cells and replacement with proliferating epithelial cells. Magnification, ×125. (b) Germinal center formation in the body mucosa and persistence of parietal cells (arrowheads). Magnification, ×50. (c) Germinal center and LEL (arrows). Magnification, ×250. (d) Massive proliferation of plasma cells in the body mucosa. Magnification, ×250. (e) Mucin-producing cells and cells which have eosinophilic cytoplasm in the corpus. Magnification, ×125. (f) Colonization of the body pits by H. pylori. Magnification, ×500. (g) Low lymphocyte infiltration in the gastric mucosa in a normal BALB/c mouse after H. pylori infection. Magnification, ×125. (h, i, and j) Massive B220⁺ cells, scattered CD4⁺ cells, and no CD8⁺ cells, respectively, in the lymph follicle of a d3-Tx mouse 4 months after H. pylori infection. Magnification, ×250. (a to e and g) Hematoxylin-eosin staining; (f) May-Grunwald Giemsa staining; (h to j) immunohistochemical staining.

FIG. 1

Sections of gastric mucosae of d3-Tx (a to f and h to j) and normal (g) BALB/c mice given saline or H. pylori orally at the age of 6 weeks. (a) Four months after saline administration; (b, c, d, f, g, h, i, and j) 4 months after H. pylori administration; (e) 12 months after H. pylori administration. (a) The body mucosa, showing the infiltration of mononuclear cells and gland atrophy characterized by loss of parietal and chief cells and replacement with proliferating epithelial cells. Magnification, ×125. (b) Germinal center formation in the body mucosa and persistence of parietal cells (arrowheads). Magnification, ×50. (c) Germinal center and LEL (arrows). Magnification, ×250. (d) Massive proliferation of plasma cells in the body mucosa. Magnification, ×250. (e) Mucin-producing cells and cells which have eosinophilic cytoplasm in the corpus. Magnification, ×125. (f) Colonization of the body pits by H. pylori. Magnification, ×500. (g) Low lymphocyte infiltration in the gastric mucosa in a normal BALB/c mouse after H. pylori infection. Magnification, ×125. (h, i, and j) Massive B220⁺ cells, scattered CD4⁺ cells, and no CD8⁺ cells, respectively, in the lymph follicle of a d3-Tx mouse 4 months after H. pylori infection. Magnification, ×250. (a to e and g) Hematoxylin-eosin staining; (f) May-Grunwald Giemsa staining; (h to j) immunohistochemical staining.

FIG. 1

Sections of gastric mucosae of d3-Tx (a to f and h to j) and normal (g) BALB/c mice given saline or H. pylori orally at the age of 6 weeks. (a) Four months after saline administration; (b, c, d, f, g, h, i, and j) 4 months after H. pylori administration; (e) 12 months after H. pylori administration. (a) The body mucosa, showing the infiltration of mononuclear cells and gland atrophy characterized by loss of parietal and chief cells and replacement with proliferating epithelial cells. Magnification, ×125. (b) Germinal center formation in the body mucosa and persistence of parietal cells (arrowheads). Magnification, ×50. (c) Germinal center and LEL (arrows). Magnification, ×250. (d) Massive proliferation of plasma cells in the body mucosa. Magnification, ×250. (e) Mucin-producing cells and cells which have eosinophilic cytoplasm in the corpus. Magnification, ×125. (f) Colonization of the body pits by H. pylori. Magnification, ×500. (g) Low lymphocyte infiltration in the gastric mucosa in a normal BALB/c mouse after H. pylori infection. Magnification, ×125. (h, i, and j) Massive B220⁺ cells, scattered CD4⁺ cells, and no CD8⁺ cells, respectively, in the lymph follicle of a d3-Tx mouse 4 months after H. pylori infection. Magnification, ×250. (a to e and g) Hematoxylin-eosin staining; (f) May-Grunwald Giemsa staining; (h to j) immunohistochemical staining.

FIG. 1

Sections of gastric mucosae of d3-Tx (a to f and h to j) and normal (g) BALB/c mice given saline or H. pylori orally at the age of 6 weeks. (a) Four months after saline administration; (b, c, d, f, g, h, i, and j) 4 months after H. pylori administration; (e) 12 months after H. pylori administration. (a) The body mucosa, showing the infiltration of mononuclear cells and gland atrophy characterized by loss of parietal and chief cells and replacement with proliferating epithelial cells. Magnification, ×125. (b) Germinal center formation in the body mucosa and persistence of parietal cells (arrowheads). Magnification, ×50. (c) Germinal center and LEL (arrows). Magnification, ×250. (d) Massive proliferation of plasma cells in the body mucosa. Magnification, ×250. (e) Mucin-producing cells and cells which have eosinophilic cytoplasm in the corpus. Magnification, ×125. (f) Colonization of the body pits by H. pylori. Magnification, ×500. (g) Low lymphocyte infiltration in the gastric mucosa in a normal BALB/c mouse after H. pylori infection. Magnification, ×125. (h, i, and j) Massive B220⁺ cells, scattered CD4⁺ cells, and no CD8⁺ cells, respectively, in the lymph follicle of a d3-Tx mouse 4 months after H. pylori infection. Magnification, ×250. (a to e and g) Hematoxylin-eosin staining; (f) May-Grunwald Giemsa staining; (h to j) immunohistochemical staining.

FIG. 1

Sections of gastric mucosae of d3-Tx (a to f and h to j) and normal (g) BALB/c mice given saline or H. pylori orally at the age of 6 weeks. (a) Four months after saline administration; (b, c, d, f, g, h, i, and j) 4 months after H. pylori administration; (e) 12 months after H. pylori administration. (a) The body mucosa, showing the infiltration of mononuclear cells and gland atrophy characterized by loss of parietal and chief cells and replacement with proliferating epithelial cells. Magnification, ×125. (b) Germinal center formation in the body mucosa and persistence of parietal cells (arrowheads). Magnification, ×50. (c) Germinal center and LEL (arrows). Magnification, ×250. (d) Massive proliferation of plasma cells in the body mucosa. Magnification, ×250. (e) Mucin-producing cells and cells which have eosinophilic cytoplasm in the corpus. Magnification, ×125. (f) Colonization of the body pits by H. pylori. Magnification, ×500. (g) Low lymphocyte infiltration in the gastric mucosa in a normal BALB/c mouse after H. pylori infection. Magnification, ×125. (h, i, and j) Massive B220⁺ cells, scattered CD4⁺ cells, and no CD8⁺ cells, respectively, in the lymph follicle of a d3-Tx mouse 4 months after H. pylori infection. Magnification, ×250. (a to e and g) Hematoxylin-eosin staining; (f) May-Grunwald Giemsa staining; (h to j) immunohistochemical staining.

FIG. 1

Sections of gastric mucosae of d3-Tx (a to f and h to j) and normal (g) BALB/c mice given saline or H. pylori orally at the age of 6 weeks. (a) Four months after saline administration; (b, c, d, f, g, h, i, and j) 4 months after H. pylori administration; (e) 12 months after H. pylori administration. (a) The body mucosa, showing the infiltration of mononuclear cells and gland atrophy characterized by loss of parietal and chief cells and replacement with proliferating epithelial cells. Magnification, ×125. (b) Germinal center formation in the body mucosa and persistence of parietal cells (arrowheads). Magnification, ×50. (c) Germinal center and LEL (arrows). Magnification, ×250. (d) Massive proliferation of plasma cells in the body mucosa. Magnification, ×250. (e) Mucin-producing cells and cells which have eosinophilic cytoplasm in the corpus. Magnification, ×125. (f) Colonization of the body pits by H. pylori. Magnification, ×500. (g) Low lymphocyte infiltration in the gastric mucosa in a normal BALB/c mouse after H. pylori infection. Magnification, ×125. (h, i, and j) Massive B220⁺ cells, scattered CD4⁺ cells, and no CD8⁺ cells, respectively, in the lymph follicle of a d3-Tx mouse 4 months after H. pylori infection. Magnification, ×250. (a to e and g) Hematoxylin-eosin staining; (f) May-Grunwald Giemsa staining; (h to j) immunohistochemical staining.

FIG. 1

Sections of gastric mucosae of d3-Tx (a to f and h to j) and normal (g) BALB/c mice given saline or H. pylori orally at the age of 6 weeks. (a) Four months after saline administration; (b, c, d, f, g, h, i, and j) 4 months after H. pylori administration; (e) 12 months after H. pylori administration. (a) The body mucosa, showing the infiltration of mononuclear cells and gland atrophy characterized by loss of parietal and chief cells and replacement with proliferating epithelial cells. Magnification, ×125. (b) Germinal center formation in the body mucosa and persistence of parietal cells (arrowheads). Magnification, ×50. (c) Germinal center and LEL (arrows). Magnification, ×250. (d) Massive proliferation of plasma cells in the body mucosa. Magnification, ×250. (e) Mucin-producing cells and cells which have eosinophilic cytoplasm in the corpus. Magnification, ×125. (f) Colonization of the body pits by H. pylori. Magnification, ×500. (g) Low lymphocyte infiltration in the gastric mucosa in a normal BALB/c mouse after H. pylori infection. Magnification, ×125. (h, i, and j) Massive B220⁺ cells, scattered CD4⁺ cells, and no CD8⁺ cells, respectively, in the lymph follicle of a d3-Tx mouse 4 months after H. pylori infection. Magnification, ×250. (a to e and g) Hematoxylin-eosin staining; (f) May-Grunwald Giemsa staining; (h to j) immunohistochemical staining.

FIG. 2

Expression of the H. pylori-urease A gene in the gastric mucosae of d3-Tx AIG [AIG(+)] and normal [AIG(−)] mice infected with H. pylori. The PCR products of extracted DNAs were analyzed by Southern blotting as described in Materials and Methods. n.c, negative control with DNA from uninfected normal mice; p.c, positive control with DNA extracted from H. pylori. Five AIG mice were analyzed individually, but the pooled gastric mucosae of three normal mice were used for the experiment. The five AIG mice were positive for the urease A gene, and the normal mice were negative.

FIG. 3

Expression of IFN-γ and IL-4 mRNAs in the gastric mucosae of d3-Tx AIG mice at 4 months after saline ingestion (a) or H. pylori (H.P.) infection (b). Saline or H. pylori was administered to each AIG mouse at the age of 6 weeks. Total RNAs were extracted from the mucosae of the five pooled stomachs, and RT-PCR was carried out as described in Materials and Methods. Five individual mice were tested for each group. n.c, negative controls without template DNA; p.c, positive controls with mRNAs isolated from Th1 clone cells (no. 16) for IFN-γ and Th2 clone cells (D10.G4.1) for IL-4. G3PDH, glyceraldehyde-3-phosphate dehydrogenase.