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Journal of Histochemistry and Cytochemistry, Vol. 49, 727-738, June 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

Transforming Growth Factor-ß Messenger RNA and Protein in Murine Colitis

Christine V. Whitinga, Amanda M. Williamsa, Mogens H. Claessonb, Soren Bregenholtb, Jorg Reimannc, and Paul W. Blanda
a Department of Clinical Veterinary Science, University of Bristol, Bristol, United Kingdom
b The Panum Institute, University of Copenhagen, Copenhagen, Denmark
c Institute for Medical Microbiology and Immunology, University of Ulm, Ulm, Germany

Correspondence to: Christine V. Whiting, Div. of Molecular & Cellular Biology, Dept. of Clinical Veterinary Science, University of Bristol, Langford, Bristol, UK BS40 5DU. E-mail: c.v.whiting@bris.ac.uk

Using a CD4+ T-cell-transplanted SCID mouse model of colitis, we have analyzed TGF-ß transcription and translation in advanced disease. By in situ hybridization, the epithelium of both control and inflamed tissues transcribed TGF-ß1 and TGF-ß3 mRNAs, but both were expressed significantly farther along the crypt axis in disease. Control lamina propria cells transcribed little TGF-ß1 or TGF-ß3 mRNA, but in inflamed tissues many cells expressed mRNA for both isoforms. No TGF-ß2 message was detected in either control or inflamed tissues. Immunohistochemistry for latent and active TGF-ß1 showed that all cells produced perinuclear latent TGF-ß1. The epithelial cell basal latent protein resulted in only low levels of subepithelial active protein, which co-localized with collagen IV and laminin in diseased and control tissue. Infiltrating cells expressed very low levels of active TGF-ß. By ELISA, very low levels (0–69 pg/mg) of soluble total or active TGF-ß were detected in hypotonic tissue lysates. TGF-ß1 and TGF-ß3 are produced by SCID mouse colon and transcription is increased in the colitis caused by transplantation of CD4+ T-cells, but this does not result in high levels of soluble active protein. Low levels of active TGF-ß may be a factor contributing to unresolved inflammation. (J Histochem Cytochem 49:727–738, 2001)

Key Words: TGF-ß, inflammatory bowel disease, in situ hybridization, basal lamina


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C. V. Whiting, J. F. Tarlton, M. Bailey, C. L. Morgan, and P. W. Bland
Abnormal Mucosal Extracellular Matrix Deposition Is Associated with Increased TGF-{beta} Receptor-expressing Mesenchymal Cells in a Mouse Model of Colitis
J. Histochem. Cytochem., September 1, 2003; 51(9): 1177 - 1189.
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