Zastosowanie metody real-time PCR oraz techniki immunohistochemicznej do oceny ekspresji receptora TLR-5 w biopsjach błony śluzowej…
The Toll-like receptor 5 (TLR-5) was found to play a role in the development of some inflammatory diseases, however, its involvement in the pathogenesis of ulcerative colitis (UC) and Crohn’s disease (CD), main entities of Inflammatory Bowel Disease (IBD) has yet to be determined. The aim of our preliminary study was to assess the expression of TLR-5 in the colonic biopsies of 2 UC and 2 CD patients by the use of real-time PCR and immunocytochemistry.
Total RNA isolated from one biopsy specimen was used to obtain cDNA by the reverse transcriptase reaction. Primers specific for exon 1 of the tlr-5 gene were used for real-time PCR reaction. In this way the initial amount of TLR-5 and β-actin mRNAs were determined for mucosal biopsies taken from inflamed and non-inflamed colonic mucosa of the same patient. We found increased TLR-5/β-actin mRNA ratio in the altered mucosa of UC patients. On the contrary, the expression of the tlr-5 gene was decreased or unchanged in the inflammatory mucosa of CD patients. The presence of TLR-5 protein was established in the enterocytes, however, no positive reaction was found in the goblet cells or lamina propria cells. The preliminary results of this quantitative and qualitative study suggest that TLR-5 expression may be different in the colonic mucosa of UC and CD patients, and that real-time PCR technique together with immunocytochemistry represent valuable methods for the assessment of TLR-5 role in the pathogenesis of IBD.
Adres: mgr inż. Maciej Wierzbicki
Katedra Histologii i Immunologii AMG
ul. Dębinki 1, 80-211 Gdańsk