Culture and functional studies of mouse macrophages on native-like fibrillar type I collagen

Article Article scientifique Article Culture and functional studies of mouse macrophages on native-like fibrillar type I collagen eng 2009 author Philippeaux Marie-Marthe 56598 author Bargetzi Jean-Pierre 25080 author Pache Jean-Claude 513830 author Robert John 55421 author Spiliopoulos Anastase 49910 author Mauel Jacques 243-256 10.1016/j.ejcb.2008.11.002 19124174 0171-9335 http://pdn.sciencedirect.com/science?_ob=MiamiImageURL&_cid=273240&_user=9565874&_pii=S0171933508001623&_check=y&_origin=article&_zone=toolbar&_coverDate=30-Apr-2009&view=c&originContentFamily=serial&wchp=dGLzVlk-zSkzS&md5=794cb6faafd4a4f6dc5988eb82568f61/1-s2.0-S0171933508001623-main.pdf Freshly isolated, starch-elicited mouse peritoneal macrophages (Mo) attached very efficiently to type I collagen in vitro, if collagen molecules were arranged in ordered supra-molecular assemblies corresponding to the precursor native fibrils. After 6-20h of incubation, the collagen-bound cells were observed to secrete fibronectin, which presumably enhanced cell-collagen interaction associated with cellular differentiation. Mo attachment to collagen could be temporarily inhibited by addition of the linear tri-peptide Arg-Gly-Asp (RGD) to the culture media. This inhibition was much more pronounced when using the cyclic RGD-containing peptide cGRGDSPA. Similarly, cells could be easily detached from the fibrillar collagen layers within 20 min at 37 degrees C by RGDS, GRGDS or cGRGDSPA but not by the glutamate-containing RGES peptide. Using antibodies to known collagen receptors, attachment of Mo to type I collagen fibers was best inhibited by antibodies directed against the alpha2 and beta1 integrin subunits. The presence of these integrins on Mo was confirmed by immunofluorescence. Binding of the alpha2beta1 integrin on collagen was divalent cation-dependent and was supported by magnesium but not by calcium. Cells recovered by RGD-mediated detachment from collagen were highly phagocytic and synthesized DNA when exposed to growth factors. These cells could be activated for cytotoxicity by treatment with interferon-gamma and lipopolysaccharide. Comparative in vitro assays performed on macrophages cultured on plastic and on collagen allowed the detection of NO production by activated macrophages followed by spontaneous deactivation for cells cultivated on collagen. These findings suggest that Mo can recognize native collagen of type I through functional interactions with their specific triple helix-binding integrin receptors indicating that integrins other than those directed to fibronectin may also occupy active focal points on the cell at the initial phase of attachment. Animals Cell Adhesion/drug effects/physiology *Cell Culture Techniques Cell Proliferation/drug effects Collagen Type I/*metabolism/ultrastructure Integrins/metabolism Interferon-gamma/pharmacology Lipopolysaccharides/pharmacology Macrophage-1 Antigen/metabolism Macrophages, Peritoneal/drug effects/metabolism/*physiology Male Mice Mice, Inbred C57BL Microscopy, Electron, Transmission Nitric Oxide/metabolism Oligopeptides/pharmacology Peptides, Cyclic/pharmacology Phagocytosis/drug effects/physiology Dewey 616.07 Dewey 617 137 Département de pathologie et immunologie 21F 57 Département de chirurgie 222 false 308 Chirurgie thoracique et chirurgie endocrinologique 573 392 Groupe Pache Jean-Claude (pathologie clinique) 658 European journal of cell biology 88 4 703d6424-6a40-4dc3-94ab-0a7833adff46 Article Restricted file1.pdf 1802328 application/pdf restrictedAccess Faculté de médecine / Section de médecine fondamentale / Département de pathologie et immunologie Faculté de médecine / Section de médecine clinique / Département de chirurgie

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Culture and functional studies of mouse macrophages on native-like fibrillar type I collagen	European journal of cell biology			2009	612	0

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Bargetzi, Jean-Pierre