title: Monocyte chemoattractant protein-1 (MCP-1) inhibits the intestinal-like differentiation of monocytes.
abstract: Monocytes (MO) migrating into normal, non-inflamed intestinal mucosa undergo a specific differentiation resulting in a non-reactive, tolerogenic intestinal macrophage (IMAC). Recently we demonstrated the differentiation of MO into an intestinal-like macrophage (MAC) phenotype in vitro in a three-dimensional cell culture model (multi-cellular spheroid or MCS model). In the mucosa of patients with inflammatory bowel disease (IBD) in addition to normal IMAC, a reactive MAC population as well as increased levels of monocyte chemoattractant protein 1 (MCP-1) is found. The aim of this study was to investigate the influence of MCP-1 on the differentiation of MO into IMAC. MCS were generated from adenovirally transfected HT-29 cells overexpressing MCP-1, macrophage inflammatory protein 3 alpha (MIP-3alpha) or non-transfected controls and co-cultured with freshly elutriated blood MO. After 7 days of co-culture MCS were harvested, and expression of the surface antigens CD33 and CD14 as well as the intracellular MAC marker CD68 was determined by flow-cytometry or immunohistochemistry. MCP-1 and MIP-3alpha expression by HT-29 cells in the MCS was increased by transfection at the time of MCS formation. In contrast to MIP-3alpha, MCP-1 overexpression induced a massive migration of MO into the three-dimensional aggregates. Differentiation of IMAC was disturbed in MCP-1-transfected MCS compared to experiments with non-transfected control aggregates, or the MIP-3alpha-transfected MCS, as indicated by high CD14 expression of MO/IMAC cultured inside the MCP-1-transfected MCS, as shown by immunohistochemistry and FACS analysis. Neutralization of MCP-1 was followed by an almost complete absence of monocyte migration into the MCS. MCP-1 induced migration of MO into three-dimensional spheroids generated from HT-29 cells and inhibited intestinal-like differentiation of blood MO into IMAC. It may be speculated that MCP-1 could play a role in the disturbed IMAC differentiation in IBD mucosa.
authors.affiliation: Department of Internal Medicine I, University of Regensburg, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Spoettl
authors.forename: M
authors.initials: M
authors.lastname: Hausmann
authors.forename: M
authors.initials: M
authors.lastname: Herlyn
authors.forename: M
authors.initials: M
authors.lastname: Gunckel
authors.forename: A
authors.initials: A
authors.lastname: Dirmeier
authors.forename: W
authors.initials: W
authors.lastname: Falk
authors.forename: H
authors.initials: H
authors.lastname: Herfarth
authors.forename: J
authors.initials: J
authors.lastname: Schoelmerich
authors.forename: G
authors.initials: G
authors.lastname: Rogler
mesh_terms: D015703:Antigens, CD / Q000032:analysis; D015214:Antigens, Differentiation, Myelomonocytic / Q000032:analysis; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects; D002465:Cell Movement; D018932:Chemokine CCL2 / Q000235:genetics / Q000378:metabolism*; D054418:Chemokine CCL20; D019742:Chemokines, CC / Q000235:genetics / Q000378:metabolism; D003864:Depression, Chemical; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D019073:HT29 Cells; D006801:Humans; D007150:Immunohistochemistry; D015212:Inflammatory Bowel Diseases / Q000276:immunology / Q000473:pathology; D007413:Intestinal Mucosa / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D019402:Macrophage Inflammatory Proteins / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000166:cytology* / Q000276:immunology; D009000:Monocytes / Q000166:cytology* / Q000276:immunology; D063268:Sialic Acid Binding Ig-like Lectin 3; D014162:Transfection / Q000379:methods; D000097382:CD68 Molecule
chemical_list: D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D015415:Biomarkers; D018932:Chemokine CCL2; D054418:Chemokine CCL20; D019742:Chemokines, CC; D018950:Lipopolysaccharide Receptors; D019402:Macrophage Inflammatory Proteins; D063268:Sialic Acid Binding Ig-like Lectin 3; C111835:CCL2 protein, human; C108355:CCL20 protein, human; C571163:CD33 protein, human; C067980:CD68 antigen, human; D000097382:CD68 Molecule