Project description:That commensal bacteria can influence intestinal inflammation has been observed using other models of chronic colitis. Loss of IL-10, a major immunosuppressive cytokine, induces spontaneous colitis in mice. The incidence of spontaneous polyp formation in IL-10-deficient mice was also completely eliminated in the absence of STING We used microarrays to evaluate the inflammatory cytokine expression in the colon from IL10 KO mice and IL10/STING KO mice.
Project description:Epithelial cells play an important role in the protection of the colon mucosa from the resident microbiota and are involved in the initiation and maintenance of intestinal inflammation. LMD is a technique that allows the extraction of specific cell types, such as colonic epithelial cells, to analyse gene expression. LMD of colon epithelial cells followed by microarray analysis could be of more value than microarray analysis of intact colon for determining which pathways are active in the colon mucosa in the early and late stages of inflammation due to increased sensitivity to changes in specific cell populations. An experiment was performed using microarray analysis of intact colon samples and microdissected colon epithelial cell samples from Il10-/- and C57BL/6J mice at 6 and 12 weeks of age to study the molecular changes that occur in early and late inflammation stages in colon epithelium of a mouse model of colitis. Results showed that intact colon and colon epithelial cell gene expression profiles were similar in terms of pathways between Il10-/- and C57BL/6J mice at 12 weeks of age and between Il10-/- mice at 12 and 6 weeks of age. More immune-related pathways were identified at 6 weeks of age in epithelial cells than intact colon. This suggests that LMD and targeting of specific cell types may be of particular use when studying the early stages of inflammation before the intestinal morphology is detectably altered, by increasing analysis sensitivity to mucosal gene expression changes. 2x2 factorial with two tissue types analysed. Two strains of mouse (Il10 knockout mouse and the background strain C57BL/6J) were sampled at 2 timepoints (6 and 12 weeks of age) and intact proximal colon and colon epithelium harvested from each mouse (6 mice per group except for group colon epithelium C57 mouse 12 weeks where only 5 samples reached quality control standards).
Project description:Lcn2 is involved in host defense against pathogens, but the function in intestinal mucosal immunity and inflammation remains largely unknown. Genetic ablation of Lcn2 results in early-onset colitis and spontaneous emergence of right-sided colonic tumors in the setting of IL-10 deficiency (Lcn2-/-;IL10-/- mice). To address whether inflammation or other mechanisms drives the site-specific tumor locations gene expression analyses in proximal versus distal colons of Lcn2-/- IL10-/- mice were performed. Differential expression between distal colon versus cecum and proximal colon samples were analyses using Affymetrix MoGene 2.0 ST arrays on formalin-fixed, paraffin-embedded tissue sections of Lcn2-/-; IL10-/-mice.
Project description:Epithelial cells play an important role in the protection of the colon mucosa from the resident microbiota and are involved in the initiation and maintenance of intestinal inflammation. LMD is a technique that allows the extraction of specific cell types, such as colonic epithelial cells, to analyse gene expression. LMD of colon epithelial cells followed by microarray analysis could be of more value than microarray analysis of intact colon for determining which pathways are active in the colon mucosa in the early and late stages of inflammation due to increased sensitivity to changes in specific cell populations. An experiment was performed using microarray analysis of intact colon samples and microdissected colon epithelial cell samples from Il10-/- and C57BL/6J mice at 6 and 12 weeks of age to study the molecular changes that occur in early and late inflammation stages in colon epithelium of a mouse model of colitis. Results showed that intact colon and colon epithelial cell gene expression profiles were similar in terms of pathways between Il10-/- and C57BL/6J mice at 12 weeks of age and between Il10-/- mice at 12 and 6 weeks of age. More immune-related pathways were identified at 6 weeks of age in epithelial cells than intact colon. This suggests that LMD and targeting of specific cell types may be of particular use when studying the early stages of inflammation before the intestinal morphology is detectably altered, by increasing analysis sensitivity to mucosal gene expression changes.
Project description:Milk and soy are reported to contain bioactive molecules with antibacterial and immunomodulatory actions, which may be beneficial to people with IBD. The aim of this study was to determine whether diets containing ruminant milk or soy solids reduce intestinal inflammation in Il10-/- mice. Male Il10-/- mice and C57BL/6J mice were fed diets containing 40% (w/w) sheep, goat, or cow whole milk powder, 40% (w/w) soy solids (NOW® Foods Soy Milk Powder, Instant), or one of two control diets (casein-free modified-AIN76A or standard AIN76A) from 4 to 11 weeks of age. Diets were based on AIN76A, which was included as an inter-experimental control for inflammation. For all diets except AIN76A, total protein, fat, carbohydrate and energy were kept as similar as possible. Weight and food intake were measured throughout the experiment (three times weekly), and intestinal tissue was taken for histopathology evaluation of inflammation and analysis of gene expression. Analysis of mouse weight and feed intake both showed a significant strain-diet interaction: Il10-/- mice fed the cow and goat milk diets ate less and gained less weight than all the other diet groups. This diet effect was not evident for the C57BL/6J mice. Il10-/- mice on the cow and goat milk diets had reduced colon histological injury scores relative to those on the other diets. Il10-/- mice on the cow and goat milk diets also had reduced expression of many immune/inflammatory-related genes and pathways.
Project description:Quantitative proteomic analysis of colon tissue extracts from CRL and IF1-KO mice was performed with isobaric tags for relative and absolute quantitation (iTRAQ) labeling method coupled to tandem mass spectrometry (MS/MS).
Project description:We performed a quantitative proteomic analysis of colon tissue extracts from CRL and IF1-KO mice with isobaric tags for relative and absolute quantitation (iTRAQ) labeling method coupled to tandem mass spectrometry (MS/MS).
Project description:We observed that deletion of polyketide synthase (pks) from E. coli NC101 reduces its ability to induce tumors in interleukin-10 knockout (Il10-/-) mice injected with azoxymethane (AOM), without altering histologic inflammation. The goal of this experiment is to assess inflammatory cytokine levels in colonic tissue of these mice. 2 germ-free Il10-/- mice were assayed and used as controls. 3 E. coli NC101 and 3 E. coli NC101-delta-pks monoassociated mice were experimental samples.
Project description:Dietary n-3 polyunsaturated fatty acids can reduce inflammation via a range of mechanisms. This study tested the effect of dietary eicosapentaenoic acid (EPA) on intestinal inflammation using interleukin-10 gene-deficient (Il10-/-) mice. Methods: At 35 days of age, 12 weaned Il10-/- and 12 C57 mice were randomly assigned to one of two modified AIN-76A diets, supplemented with 3.7% purified ethyl esters of either EPA (n-3) or oleic acid (OA, control). To identify genes relevant to colon inflammation, transcription profiling (microarrays and qRT-PCR) and bioinformatic analyses were used. Results: In this study, dietary EPA reversed the decrease in colon fatty acid β-oxidation gene expression observed in OA-fed Il10-/- compared to C57 mice. Il10-/- mice fed the OA diet showed decreased expression of antioxidant enzyme genes, as well as those involved in detoxification of xenobiotics, compared to C57 mice on the same diet. In contrast, dietary EPA increased the expression of these genes in Il10-/- mice. Conclusions: These data indicate that dietary EPA induced endogenous lipid oxidation which might have a potential anti-inflammatory effect on colon tissue. This is supported by the activation of the Ppara gene that regulates the expression of pro-inflammatory and immunomodulatory genes and proteins.
Project description:We observed that interleukin-10 knockout (Il10-/-) mice injected with azoxymethane (AOM) and monoassociated with E. coli NC101, but not E. faecalis OG1RF, develop tumors. Histologic inflammation is not different in mice monoassociated with either bacterium. The goal of this experiment is to assess inflammatory cytokine levels in colonic tissue of these mice. 4 germ-free Il10-/- mice were assayed and used as controls. 4 E. coli and 4 E. faecalis monoassociated mice were experimental samples.