부산대학교 도서관

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.vetimm.2006.06.010 Byline: N. Nguyen Van, K. Taglinger, C.R. Helps, S. Tasker, T.J. Gruffydd-Jones, M.J. Day Keywords: Cat; Cytokine; Intestine Abbreviations: gDNA, genomic DNA; IBD, inflammatory bowel disease; CD, Crohn's disease; UC, ulcerative colitis; C.sub.t, threshold cycle; GI, gastrointestinal; mRNA, messenger RNA; RT-PCR, reverse transcriptase polymerase chain reaction; SPF, specific pathogen free Abstract: Inflammatory bowel disease (IBD) is a common condition in cats characterised by infiltration of inflammatory cells into the intestinal mucosa. In this study, real-time reverse transcriptase polymerase chain reaction (RT-PCR) was used to quantify cytokine messenger RNA (mRNA) expression in intestinal biopsies from cats. Biopsies were collected from seven cats with chronic diarrhoea and histologically confirmed IBD, five cats with chronic diarrhoea due to non-IBD gastrointestinal (GI) disease, and nine clinically normal cats with or without subclinical inflammatory changes in small intestine. Real-time RT-PCR was developed for quantification of mRNA encoding interleukin (IL)-2, IL-4, IL-5, IL-6, IL-10, IL-12 (p35 and p40), IL-18, tumour necrosis factor-alpha (TNF-[alpha]), interferon-gamma (IFN-[gamma]) and transforming growth factor-beta (TGF-[beta]). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a 'housekeeper' gene. All real-time PCR efficiencies were90% (range 90.4-102%) with correlation coefficients >0.99 (range 0.998-1). The results of the study were analyzed on the basis of either clinical presentation or histopathological evidence of intestinal inflammation. The former analysis showed that mRNA encoding IL-10 and TGF-[beta] (immunoregulatory cytokines), and IL-6, IL-18, TNF-[alpha] and IL-12 p40 (Th1 and pro-inflammatory cytokines) was significantly higher in clinically normal cats and cats with IBD when compared to cats with other GI diseases. IL-5 mRNA was significantly higher in cats with IBD compared to clinically normal cats. IL-2 mRNA was significantly lower in cats with non-IBD GI disease than in clinically normal cats. Analysis on the basis of histopathological change revealed that cats with intestinal inflammation had significantly more transcription of genes encoding IL-6, IL-10, IL-12p40, TNF-[alpha] and TGF-[beta] than those with normal intestinal morphology. The results suggest that immune dysregulation plays a role in feline IBD and that IBD in cats has a complicated pathogenesis with both pro-inflammatory and immunoregulatory features. Author Affiliation: School of Clinical Veterinary Science, University of Bristol, Langford, Bristol BS40 5DU, United Kingdom Article History: Received 20 March 2006; Revised 9 June 2006; Accepted 26 June 2006