1. Volume 133, Issue 4, Pages 1210-1218 (October 2007)
Muc1 Mucin Limits Both Helicobacter pylori Colonization of the Murine Gastric Mucosa and Associated Gastritis 
Michael A. McGuckin, Alison L. Every, Caroline D. Skene, Sara K. Linden, Yok Teng Chionh, Agnieszka Swierczak, Julie McAuley, Stacey Harbour, Maria Kaparakis, Richard Ferrero, Philip Sutton 
Gastroenterology 
Volume 133, Issue 4, Pages (October 2007)
DOI: /j.gastro
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2. Figure 1
H pylori colonization levels are chronically elevated in Muc1−/− mice compared with wild-type controls. Wild-type (Muc1+/+) or Muc1−/− mice were infected with a single challenge of H pylori-SS1. At each time point, stomachs were removed, and bacterial colonization was determined by colony-forming assay. Data shown are pooled from 3 separate experiments. Box plots present the median colony-forming units (horizontal bar), interquartile range (boxed region), and the 10th and 90th percentile values (error bars). *Colonization in Muc1−/− was significantly higher than wild-type controls at the same time point (Mann–Whitney U test; P < .05). Numbers in parentheses indicate the fold increase in median bacterial colonization in Muc1−/− mice compared with wild-type controls.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

3. Figure 2
Heterozygous Muc1+/− mice exhibit intermediate H pylori colonization levels. Muc1+/+, Muc1+/−, or Muc1−/− mice were infected with a single challenge of H pylori-SS1. Two weeks postchallenge, stomachs were removed, and bacterial colonization levels determined by colony-forming assay. Box plots present the median colony-forming units (horizontal bar), interquartile range (boxed region), and the 10th and 90th percentile values (error bars). *Colonization in Muc1−/− was significantly higher than both Muc1+/− and Muc1+/+ mice (Mann–Whitney U test; P < .016). There was also a clear trend for higher colonization in the heterozygous mice compared with wild-type mice (Mann–Whitney U test; P < .065). Numbers in parentheses indicate the fold increase in median bacterial colonization in Muc1+/− and Muc1−/− mice compared with wild-type controls.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

4. Figure 3
Heterozygous Muc1+/− mice exhibit intermediate expression of Muc1 in the gastric mucosa. Gastric mucosal protein extracts from Muc1−/−, Muc1+/−, and Muc1+/+ mice (n = 3 pooled samples per strain) were separated by SDS-PAGE and subjected to Western blotting for detection of Muc1 with the monoclonal antibody CT2. CT2 binds the α subunit of Muc1, which under SDS-PAGE appears as a broad series of bands at ∼25 kilodaltons depending on glycosylation.31
Gastroenterology  , DOI: ( /j.gastro )
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5. Figure 4
Histopathology in Muc1−/− and wild-type mice following 2 months infection with H pylori. Images were taken from H&E-stained gastric sections, viewed under light microscopy (10× objective), and are to scale. (A) Corpus from a typical Muc1+/+ mouse infected with H pylori for 2 months, with a normal morphology except for a low level cellular infiltrate (CI). This level of inflammation is the typical mild gastritis observed in other mouse strains infected with H pylori.5 CI in this mouse was graded as 1, and all other parameters were zero. (B) Corpus from a typical Muc1−/− mouse infected with H pylori for 2 months. The gastric mucosa from these mice shows a marked cellular infiltrate both above (CI) and below (submucosal inflammation; SMI) the mucosae muscularis. The gastric sections from these mice display mucosal thickening with crypt elongation, loss of parietal cells, and abscesses (Ab). The histopathology in this mouse was graded as CI, 3; SMI, 3; and atrophy, 2.
Gastroenterology  , DOI: ( /j.gastro )
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6. Figure 5
Muc1-deficient mice exhibit normal immune responsiveness to infection and mitogen. Muc1−/− and Muc1+/+ mice were infected with H pylori-SS1 for 2 months before collection of spleens and sera. (A) Splenocytes were stimulated in vitro with H pylori lysate or Con A for 2 days then pulsed with 3H-thymidine. Data are presented as the mean counts per minute (cpm) of 6 replicates ± SD. (B) Antibody levels against H pylori in sera were compared by ELISA and expressed as end point titer minus background titers from uninfected controls. Box plots present the median colony-forming units (horizontal bar), interquartile range (boxed region), and the 10th and 90th percentile values (error bars). There was no significant difference in the proliferative or antibody responses of the Muc1−/− and wild-type mice.
Gastroenterology  , DOI: ( /j.gastro )
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7. Figure 6
H pylori binds to human MUC1. MUC1 from the cellular lysate of the MKN7 gastric cancer cell line was captured by a microtiter plate coated with antibody against MUC1. Control wells were coated with an isotype control. Biotinylated H pylori strains SS1 and J99 were added to the wells, and binding was detected with streptavidin-HRP. Data presented are mean values of 6 replicates ± SD after subtraction of the mean value of the isotype control for that sample. *MUC1 capture vs isotype control, P < .001 (Student t test).
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

8. Figure 7
Increased adherence of H pylori to murine primary gastric epithelial cells, which lack Muc1. Primary epithelial cells from Muc1−/− and Muc1+/+ littermates (n = 4) were cultured in 96-well plates for 2 days. (A) H pylori were added to 6 wells per mouse, with another 6 wells left with no bacteria. Positive controls: wells with no gastric cells, to which bacteria were added and allowed to adhere to the plastic before fixation. Negative controls: wells with no gastric cells or bacteria. Bacterial adhesion to cultured cells was determined by ELISA. Data shown are mean percentage adherence (calculated from: [(OD gastric cells with H pylori − OD gastric cells without H pylori)/(OD positive control − OD negative control)] × 100) ± SD. *Significantly greater bacterial adhesion compared with wild-type controls, P < .001 (Student t test). (B) Primary cell lysates from 2 wild-type and 2 Muc1−/− mice were run separately on SDS-PAGE and proteins transferred to nitrocellulose, which was then probed with anti-Muc1 antibody and detected using biotinylated anti-hamster, streptavidin-HRP, and ECL.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions