MMP dependence of fibroblast contraction and collagen production induced by human mast cell activation in a three-dimensional collagen lattice

Abstract

Mast cell-fibroblast interactions may contribute to fibrosis in asthma and other disease states. Fibroblast contraction is known to be stimulated by coculture with the human mast cell line, HMC-1, or by mast cell-derived agents. Matrix metalloproteinases (MMPs) can also mediate contraction, but the MMP-dependence of mast cell-induced fibroblast contractility is not established, and the consequences of mast cell activation within the coculture system have not been fully explored. We demonstrate that activation of primary human mast cells (pHMC) with IgE receptor cross-linking, or activation of HMC-1 with C5a, enhanced contractility of human lung fibroblasts in a three-dimensional collagen lattice system. This enhanced contractility was inhibited by the pan-MMP antagonist, batimastat, and was transferrable in the conditioned medium of activated mast cells. Exogenously added MMPs promoted gel contraction by mediating the proteolytic activation of latent transforming growth factor-beta (TGF-beta). Consistent with this, fibroblast contraction induced by mast cell activation was enhanced by addition of excess latent TGF-beta to the cultures. Batimastat inhibited this response, suggesting that MMPs capable of activating latent TGF-beta were released following mast cell activation in coculture with fibroblasts. Collagen production was also stimulated by activated mast cells in an MMP-dependent manner. MMP-2 and MMP-3 content of the gels increased in the presence of activated mast cells, and inhibition of these enzymes blocked the contractile response. These findings demonstrate the MMP dependence of mast cell-induced fibroblast contraction and collagen production.

IL-33 synergizes with IgE-dependent and IgE-independent agents to promote mast cell and basophil activation

“…To generate human primary mast cells, CD34-positive progenitors were purified from PBMC, as previously described [41]. Cells were cultured in Iscove's medium containing glutamine (Lonza, Walkersville, MD, USA), 100 ng/ml SCF (R&D Systems, Minneapolis, MN, USA), 3% 209 concentrated conditioned medium generated from HCC2157 BL lymphoblastoid cell line (ATCC), and 10 pg/ml GM-CSF (R&D Systems) at 10 4 cells/ml.…”

Mast Cell-Dependent Contraction of Human Airway Smooth Muscle Cell-Containing Collagen Gels: Influence of Cytokines, Matrix Metalloproteases, and Serine Proteases

“…We have recently used a contraction model with collagen gels coembedded with human lung fibroblasts and mast cells to study cellular interactions contributing to fibrosis (18). We now adapt this system to model mast cell interactions with human airway smooth muscle cells (HASM) and explore the consequences of human mast cell activation in coculture with smooth muscle cells in the collagen gel matrix.…”

Characterization of Total and Active Matrix Metalloproteinases-1, -3, and -13 Synthesized and Secreted by Anterior Cruciate Ligament Fibroblasts in Three-Dimensional Collagen Gels

“…[20][21][22][23][24][25] In this study, the MMP production and activity was measured in cells cultured in 3D collagen gels under static conditions, therefore no extrinsic load was applied to the gels. Gels remained tethered throughout the experiment.…”

“…Several studies have suggested that general MMP inhibitors can prevent fibroblast-induced gel contraction. 20,24,28,29 Although specific MMPs involved have not been identified, MMP-2 and -9 have been implicated. 30,31 Other studies have suggested that fibroblasts exhibit contractile force independent of MMPs 32 through alteration in the cytoskeleton and cell-cell interactions.…”