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(American Journal of Pathology. 1999;154:105-111.)
© 1999 American Society for Investigative Pathology

Technical Advances

Active Transforming Growth Factor-ß in Wound Repair

Determination Using a New Assay

Liju Yang, Cindy X. Qiu, Anna Ludlow, Mark W. J. Ferguson and Georg Brunner

From the Division of Cells, Immunology and Development, School of Biological Sciences, University of Manchester, Manchester, United Kingdom

Transforming growth factor (TGF)-ß regulates wound repair and scarring in an isoform-specific fashion. TGF-ß is produced in a latent form, and its activation is a critical regulatory step controlling the bioactivity of this growth factor. To date, it has been impossible to determine latent TGF-ß activation in vivo due to a lack of quantitative assays. We describe here a semiquantitative modification of the plasminogen activator inhibitor-1/luciferase bioassay (PAI/L assay) for TGF-ß, which we used to determine active and latent TGF-ß isoforms in frozen sections of rat wound tissue. We found that significant amounts of latent TGF-ß were rapidly activated upon wounding (38% of the total TGF-ß at 1 hour after wounding). A second peak of active TGF-ß (17% of total) occurred at 5 days after wounding. The predominant isoforms were TGF-ß1 and -2 with only minor amounts of TGF-ß3 present. This is the first TGF-ß bioassay allowing semiquantitative determination of active and latent isoforms present in vivo, and our results document the significance and temporal regulation of latent TGF-ß isoform activation in wound repair.




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