1. Volume 128, Issue 7, Pages 1937-1952 (June 2005)
Helicobacter felis Eradication Restores Normal Architecture and Inhibits Gastric Cancer Progression in C57BL/6 Mice 
Xun Cai, Jane Carlson, Calin Stoicov, Hanchen Li, Timothy C. Wang, JeanMarie Houghton 
Gastroenterology 
Volume 128, Issue 7, Pages (June 2005)
DOI: /j.gastro
Copyright © 2005 American Gastroenterological Association Terms and Conditions

2. Figure 1
Eradication protocol. (A) Mice were infected and received bacterial eradication therapy at 2, 6, or 12 months or were left infected and killed at set time points after eradication as shown by the arrows. The black bar represents the length of infection, and the white bar represents noninfection. (B) flaB polymerase chain reaction was used to confirm infection status, with glyceraldehyde-3-phosphate dehydrogenase as eukaryotic DNA control. All mice were evaluated, and a representative gel is shown.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

3. Figure 2
Natural progression of H felis infection in the C57BL/6 mouse model. (A–C) Sham infected. (A) Fundic mucosal strip taken from the lesser curvature at the squamocolumnar junction. (B) Antral mucosa. (C) IHC for H+,K+-ATPase. The arrow indicates large parietal with central nuclei and brown staining cytoplasm. (D–F) H felis infection for 4 months. (D) Fundic mucosal strip taken from the lesser curvature at the squamocolumnar junction. Moderate to severe submucosal chronic inflammatory infiltrate (thin arrow) and intramucosal chronic inflammatory cells (thick arrows) and antralization of glands (bar) are seen. (E) Antral mucosa with intramucosal inflammatory infiltrate (arrow) and mucous metaplasia of glands (bar). (F) IHC for H+,K+-ATPase. (G–I) H felis infection for 8 months. (G) Fundic submucosal chronic inflammatory infiltrates (thick arrow), intramucosal inflammation, marked mucous cell metaplasia (thin arrow), and complete loss of parietal cells are prominent. (H) Antral mucosa. (I) IHC for H+,K+-ATPase confirms a complete absence of parietal cells. (J–L) H felis infection for 14 months. (J) Dysplastic dilated cystic fundic gastric glands (gastrointestinal intraepithelial neoplasia) (thin arrow), and dilated cystic glands invading into the submucosal layer surrounded by chronic inflammatory cells (thick arrow). (K) Markedly thickened antral mucosa comprised of dysplastic glands (surface) and mucous metaplasia of deeper glands (bar). (L) IHC for H+,K+-ATPase confirms complete loss of parietal cells. (M–O) At 18 months postinfection, there is progressive architectural distortion of the fundus (M) with dilated cystic glands both intramucosal (thin arrow) and submucosal (thick arrow). (N) Architectural distortion of the antral mucosa is progressive with severe metaplasia and dysplasia of glands, profound thickening, and loss of all normal gland structure. (O) IHC for H+,K+-ATPase confirms a complete absence of parietal cells. (P) The fundus at 22 months of infection contains bizarre dilated cystic glands invading into the submucosal layer, surrounded by chronic inflammatory cells. The arrow indicates the squamocolumnar junction. (Q) Frank adenocarcinoma of the antrum. Cells have nuclear to cytoplasmic ratio alterations, loss of nuclear polarity, and architectural distortion. (R) Gross pathology at 22 weeks: (1) squamous forestomach; (2) fundus, greater curvature; (3) fundus, lesser curvature, more pronounced mucosal thickening; (4) large antral polypoid lesion causing gastric outlet obstruction; (5) proximal duodenum. All sections of fundus were taken at the squamocolumnar junction. Fundus and antral histology; H&E stain, original magnification 200×. H+,K+-ATPase IHC counterstained with hematoxylin.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

4. Figure 3
Histology scores. Mice were sham infected or infected with H felis and killed at the time points indicated. Inflammation, hyperplasia, and dysplasia were scored on a 0–4 scale. Two sections through the squamocolumnar junction at the lesser curvature were evaluated for each mouse, and the worst score was recorded for each section evaluated. (A) Natural history of H felis infection from 4 to 22 months of infection. (B) Mice infected with H felis for 2 months received sham eradication therapy or effective eradication therapy and were killed after 2, 6, or 12 additional months. (C) Mice infected with H felis for 6 months received sham eradication therapy or effective eradication therapy and were killed after 2, 6, or 12 additional months. (D) Mice infected with H felis for 12 months received sham eradication therapy or effective eradication therapy and were killed after 2, 6, or 12 additional months. n = 5 for each group. All experimental groups are compared with infected controls. A horizontal bar extending over more than one group indicates that each group was independently compared with infected control and shares a P value. Data are reported as mean ± SD. *P < .05.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

5. Figure 4
Eradication of Helicobacter infection restores parietal cell mass. Mice at early (2 months), mid (6 months), or late (12 months) points of infection received triple eradication therapy and were killed 2, 6 (data not shown), or 12 months later. (A) Sham eradication therapy at 2 months of infection. (B and C) Two months of infection, followed by eradication therapy, and killed (B) 2 months later or (C) 12 months later shows complete restoration of architecture and regression of inflammatory cell infiltrates. (D) H+,K+-ATPase IHC demonstrates complete restoration of parietal cell mass at 12 months after eradication therapy. (E) Six months of infection, sham eradication therapy. (F and G) Six months of infection, followed by eradication therapy, and killed (F) 2 months later or (G) 12 months later. Complete restoration of architecture is noted, with mild residual inflammation at 2 months and complete regression of inflammation at 12 months. (H) H+,K+-ATPase IHC shows complete restoration of parietal cell mass at 12 months posteradication. (I) Twelve months of infection, sham eradication therapy. (J and K) Twelve months of infection, followed by eradication therapy, and killed (J) 2 months later or (K) 12 months later. (L) H+,K+ATPase IHC shows partial restoration of parietal cell mass.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

6. Figure 5
Bacterial eradication at 1 year of infection prevents death due to obstructing antral adenocarcinoma/gastric outlet obstruction. (A) Mice were infected with H felis, received sham eradication or eradication therapy at 12 months of infection, and were followed up until 26 months. At 24 months of infection, all infected mice had died; at 26 months (14 months after eradication therapy), all mice that had received eradication therapy were alive. (B) Incidence of microscopic invasive antral adenocarcinoma at 8, 14, 16, and 22 months of infection and in mice infected for 12 months that received bacterial eradication and were evaluated at 24 months (12 months after eradication therapy). For 8, 14, and 16 months of infection, n = 5; for 22 and 24 months of infection, n = 10.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

7. Figure 6
Bacterial eradication decreases gastric mucosal proliferation. Mice infected with H felis for 2, 6, or 12 months received bacterial eradication and were killed 2 or 12 months later. All mice received intraperitoneal BrdU 1 hour before they were killed. (A–I) Fundus. (J–R) Antrum. BrdU LI counted as the number of positive nuclei per 20 glands. (A) Mice were infected for 2 months (early infection) and killed 2 months later (4-month time point) or 12 months later (14-month time point). IHC for BrdU in (B) eradicated and (C) uneradicated mice. (D) Mice were infected for 6 months (midpoint), received eradication therapy, and were killed 2 months later (8-month time point) or 12 months later (18-month time point). Representative IHC for BrdU in (E) eradicated and (F) uneradicated mice are shown. (G) Mice were infected with H felis for 12 months (late infection), received eradication therapy, and were killed 2 months later (14-month time point) or 10 months later (22-month time point). IHC for BrdU in (H) uneradicated and (I) eradicated mice are shown. BrdU LI in the antrum for (J–L) early, (M–O) mid, and (P–R) late time points of infection as described for the corresponding sections of fundus described previously. n = 5 mice per group. Twenty glands were counted per histologic section. Two histologic sections were counted per mouse and reported as the average ± SD. Results were considered statistically significant at P < .05.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

8. Figure 7
α-catenin expression pattern is restored with successful bacterial eradication. α-catenin expression was evaluated by IHC (brown staining), counterstained with hematoxylin (blue). (A) Control mice. Mice infected wiwth H felis for (B) 2 months, (C) 6 months, (D) 12 months, and (E) 24 months. Short-term effects of eradication therapy on α-catenin were evaluated in tissues 2 months after sham or effective eradication therapy at (F and G) 2 months, (H and I) 6 months, or (J and K) 12 months of infection. Mice eradicated at 12 months were evaluated at a late time point of 22 months (10 months posteradication, L) along with (M) infected control. (A–M, original magnification 400×.)
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

9. Figure 8
β-catenin expression pattern is restored with successful bacterial eradication. β-catenin expression was evaluated by IHC (brown staining), counterstained with hematoxylin (blue). (A) Control mice. Mice infected with H felis for (B) 2 months, (C) 6 months, (D) 12 months, and (E) 24 months. Short-term effects of eradication therapy on β-catenin were evaluated in tissues 2 months after sham or effective eradication therapy at (F and G) 2 months, (H and I) 6 months, or (J and K) 12 months of infection. Mice eradicated at 12 months were evaluated at a late time point of 22 months (10 months posteradication, L) along with (M) infected control. (A–M, original magnification 400×.)
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

10. Figure 9
IQGAP1 expression pattern is restored with successful bacterial eradication. IQGAP1 expression was evaluated by IHC (brown staining), counterstained with hematoxylin (blue). (A) Control mice. Mice infected with H felis for (B) 2 months, (C) 6 months, (D) 12 months, and (E) 24 months. Short-term effects of eradication therapy on IQGAP1 were evaluated in tissues 2 months after sham or effective eradication therapy at (F and G) 2 months, (H and I) 6 months, or (J and K) 12 months of infection. Mice eradicated at 12 months were evaluated at a late time point of 22 months (10 months posteradication, L) along with (M) infected control. (A–M, original magnification 400×.)
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions