THROMBOSPONDIN CAUSES ACTIVATION OF LATENT TRANSFORMING GROWTH-FACTOR-BETA SECRETED BY ENDOTHELIAL-CELLS BY A NOVEL MECHANISM

Thrombospondin (TSP) forms specific complexes with transforming growth factor-beta (TGF-beta) in the alpha granule releasate of platelets an d these TSP-TGF-beta complexes inhibit the growth of bovine aortic end othelial cells (BAE). In these studies, we report that TSP stripped of associated TGF-beta (sTSP) retained growth inhibitory activity which was partially reversed by a neutralizing antibody specific for TGF-bet a. Since BAE cells secrete latent TGF-beta, we determined whether sTSP activates the latent TGF-beta secreted by BAE cells. Cells were cultu red with or without sTSP and then the conditioned medium was tested fo r the ability to support TGF-beta-dependent normal rat kidney (NRK) co lony formation in soft agar. Medium conditioned with sTSP showed a dos e- and time-dependent ability to stimulate BAE-secreted TGF-beta activ ity, reaching maximal activation by 1-2 h with 0.4 mug/ml (0.9 nM) sTS P. The sTSP-mediated stimulation of TGF-beta activity is not dependent on serum factors and is not a general property of extracellular matri x molecules. The sTSP-mediated stimulation of TGF-beta activity was bl ocked by a mAb specific for sTSP and by neutralizing antibodies to TGF -beta. Activation of BAE cell secreted latent TGF-beta by sTSP can occ ur in the absence of cells and apparently does not require interaction s with cell surface molecules, since in conditioned medium removed fro m cells and then incubated with sTSP, activation occurs with kinetics and at levels similar to what is seen when sTSP is incubated in the pr esence of cells. Serine proteases such as plasmin are not involved in sTSP-mediated activation of TGF-beta. Factors that regulate the conver sion of latent to active TGF-beta are keys to controlling TGF-beta act ivity. These data suggest that TSP is a potent physiologic regulator o f TGF-beta activation.