Interactions between monocytes and smooth-muscle cells can lead to extracellular matrix degradation Yun Kui Zhu, MDa, Xiangde Liu, MDb, Hangjun Wang,

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Interactions between monocytes and smooth-muscle cells can lead to extracellular matrix degradation Yun Kui Zhu, MDa, Xiangde Liu, MDb, Hangjun Wang, MDc, Tadashi Kohyama, MDb, Fu-Qiang Wen, MD, PhDb, C.Magnus Sköld, MD, PhDd, Stephen I. Rennard, MDb Journal of Allergy and Clinical Immunology Volume 108, Issue 6, Pages 989-996 (December 2001) DOI: 10.1067/mai.2001.120193 Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 1 Effect of NE on SMCs cocultured with blood monocytes on collagen degradation. Gels were cultured for 5 days, and hydroxyproline content was determined as described in the “Methods” section. Vertical axis , Hydroxyproline content in the gels (percentage to SMC alone); horizontal axis , culture conditions. Data are means ± SEM of triplicate samples from 3 separate experiments. *P < .01 by t test and P < .01 by Tukey correction compared with SMCs alone. BM , Blood monocytes. Journal of Allergy and Clinical Immunology 2001 108, 989-996DOI: (10.1067/mai.2001.120193) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 2 TNF-α and IL-1β release. Cells were cultured in collagen gels, and media were collected on day 5. TNF-α and IL-1β were determined with ELISA. Vertical axis , Cytokine concentrations (in nanograms per milliliter); horizontal axis , culture conditions. BM , Blood monocytes. Journal of Allergy and Clinical Immunology 2001 108, 989-996DOI: (10.1067/mai.2001.120193) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 3 Effects of exogenous TNF-α and IL-1β on collagen degradation mediated by SMCs. SMCs were cultured in collagen gels and floated in media containing TNF-α (10 ng/mL) and IL-1β (5 ng/mL) or combined with NE (20 nmol/L). Hydroxyproline content in the gels was determined on day 5. Vertical axis , Hydroxy-proline content (percentage to control); horizontal axis , culture conditions. Data are means ± SEM of triplicate samples from 3 separate experiments. *P < .01 by Tukey correction compared with control. Journal of Allergy and Clinical Immunology 2001 108, 989-996DOI: (10.1067/mai.2001.120193) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 4 PGE2 release in cocultures. Gels were released into media containing NE (20 nmol/L), indomethacin (1 μmol/L), or both. After 5 days, PGE2 in the medium was quantified by means of enzyme immunoassay. Vertical axis , PGE2 concentration; horizontal axis , culture condition. *P < .01 by Tukey correction compared with SMCs alone. INDO , Indomethacin; BM , blood monocytes. Journal of Allergy and Clinical Immunology 2001 108, 989-996DOI: (10.1067/mai.2001.120193) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 5 The effect of PGE2 on collagen degradation. Gels were released into media containing NE (20 nmol/L), indomethacin (1 μmol/L), PGE2 (0.1 μmol/L), or some combination thereof. Hydroxyproline was quantified on day 5. Vertical axis , Hydroxyproline (percentage to SMCs alone); horizontal axis , culture conditions. The data are means ± SEM of triplicate samples from 3 separate experiments. *P < .01 by Tukey correction. INDO , Indomethacin; BM , blood monocytes. Journal of Allergy and Clinical Immunology 2001 108, 989-996DOI: (10.1067/mai.2001.120193) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 6 Effect of dexamethasone on collagen degradation. Gels were released into media containing NE (20 nmol/L), dexamethasone (1 μmol/L), or both, and hydroxyproline was quantified on day 5. Vertical axis , Hydroxyproline (percentage to SMCs alone); horizontal axis , culture conditions. The data are means ± SEM of triplicate samples from 2 separate experiments. *P < .01 by Tukey correction. DEX , Dexamethasone; BM , blood monocytes. Journal of Allergy and Clinical Immunology 2001 108, 989-996DOI: (10.1067/mai.2001.120193) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 7 MMP-2 and MMP-9 activity. Gelatin zymography was used to identify gelatinases in various media. Additions were NE (20 nmol/L), indomethacin (INDO ; 1 μmol/L), dexamethasone (DEX ; 1 μmol/L), PGE2 (0.1 μmol/L), or some combination thereof. Molecular mass markers are indicated. Journal of Allergy and Clinical Immunology 2001 108, 989-996DOI: (10.1067/mai.2001.120193) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 8 MMP-1 detection. Immunoblotting was used to detect MMP-1 in various media. Additions were NE (20 nmol/L), indomethacin (INDO ; 1 mmol/L), dexamethasone (DEX ; 1 μmol/L), PGE2 (0.1 μmol/L), or some combination thereof. Molecular mass markers are indicated. Journal of Allergy and Clinical Immunology 2001 108, 989-996DOI: (10.1067/mai.2001.120193) Copyright © 2001 Mosby, Inc. Terms and Conditions