Measurement of cytokine mRNA expression in intestinal biopsies of cats with inflammatory enteropathy using quantitative real-time RT-PCR

Văn, Nghĩa Nguyễn; Taglinger, K; Helps, Christopher R; Tasker, Séverine; Gruffydd-Jones, TJ; Day, Michael

doi:10.1016/j.vetimm.2006.06.010

Cited by 38 publications

(24 citation statements)

References 22 publications

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“…The absence of genomic contamination of all RNA samples was confirmed before the RT reactions by performing qPCR on the purified RNA without reverse transcription with the G3PDH assay as previously described 6. None of the samples showed evidence of amplifiable genomic DNA with this assay.…”

Section: Methodsmentioning

confidence: 90%

“…Isolation and DNase digestion of total RNA from 2 to 4 endoscopic biopsy specimens (8–18 mg) were performed as described previously 6. The quantity and quality of the resulting total RNA were assessed by automated gel electrophoresis 2…”

Section: Methodsmentioning

confidence: 99%

“…6 No feline‐specific IL‐12p19 sequence was available from GenBank for the design of primers and probes for this target. Therefore, a consensus sequence was obtained by aligning the human (GenBank accession number: NM_016584), murine (NM_031252) and canine (XM_538231) sequences.…”

Section: Methodsmentioning

confidence: 99%

“…Characterization of mucosal cellular infiltrates using immunohistochemical techniques in canine IBD2, 3 and in healthy cats and cats with IBD4, 5 has provided new insights into the cellular changes that may accompany the disease process. Recent reports have shown alterations in cytokine gene expression profiles in the small intestine of cats with IBD 6, 7. One of the key cytokines with a role in the immunopathogenesis of IBD is interleukin‐12 (IL‐12) which is a heterodimeric molecule composed of p35 and p40 subunits and which promotes development of IFN‐γ‐producing Th1 CD4 + cells 8…”

mentioning

confidence: 99%

“…The role of IL‐12 in the pathogenesis of IBD is being re‐evaluated in light of recent reports suggesting that IL‐23 may have greater relevance to the pathogenesis of various autoimmune and inflammatory diseases 10. Significant differences have been reported in the levels of expression of mRNA encoding IL‐12p40, but not IL‐12p35 between cats with and without histologic evidence of IBD 6. In light of the new findings related to IL‐23, up‐regulation of p40 mRNA may be associated with parallel up‐regulation of IL‐23p19 gene expression.…”

mentioning

confidence: 99%

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Measurement of IL‐12 (p40, p35), IL‐23p19, and IFN‐γ mRNA in Duodenal Biopsies of Cats with Inflammatory Enteropathy

Waly

Peters

Day

et al. 2013

Veterinary Internal Medicne

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BackgroundDietary hypersensitivity and inflammatory bowel disease (IBD) are important causes of chronic vomiting and diarrhea in cats. IL‐23 has been recently found to be a key factor in the immunopathogenesis of IBD in humans but the involvement in IBD has not been investigated in cats.Hypothesis/ObjectivesExpression of genes encoding Il‐12p35 and p40, IL‐23p19, and IFN‐γ may be up‐regulated in duodenal biopsy specimens taken from cats with histologic evidence of inflammation.Animals and MethodsDuodenal biopsy specimens were collected from control cats (n = 21) and cats with inflammatory enteropathy (n = 13). Routine histopathology, immunohistochemistry (IHC), and qRT‐PCR were used to assess expression of MHC class II and to measure gene transcripts encoding the p35, p40, and p19 subunits of the IL‐12 family of cytokines and IFN‐γ.ResultsThere were significant differences in expression of mRNA encoding IL‐12p35 and IL‐23p19 between healthy cats and cats with inflammatory enteropathy. IL‐12p35 mRNA was lower in the duodenal mucosa of cats with inflammatory enteropathy compared with the mucosa of healthy cats (P = .001). In contrast, IL‐23p19 mRNA expression was higher in duodenal biopsy specimens from cats with inflammatory enteropathy than in those from healthy controls (P = .001). There was no difference in expression of IL‐12p40 and IFN‐γ mRNA (P > .05). The majority of cats with inflammatory enteropathy had histologic evidence of moderate to severe colitis (score 2).Conclusions and Clinical ImportanceThe results of this preliminary study suggest that IL‐23 plays a role in the pathogenesis of feline inflammatory enteropathy.

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Section: Methodsmentioning

confidence: 90%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%