title: IL-10 overexpression differentially affects cartilage matrix gene expression in response to TNF-alpha in human articular chondrocytes in vitro.
abstract: Cartilage-specific extracellular matrix synthesis is the prerequisite for chondrocyte survival and cartilage function, but is affected by the pro-inflammatory cytokine TNF-alpha in arthritis. The aim of the present study was to characterize whether the immunoregulatory cytokine IL-10 might modulate cartilage matrix and cytokine expression in response to TNF-alpha. Primary human articular chondrocytes were treated with either recombinant IL-10, TNF-alpha or a combination of both (at 10ng/mL each) or transduced with an adenoviral vector overexpressing human IL-10 and subsequently stimulated with 10ng/ml TNF-alpha for 6 or 24h. The effects of IL-10 on the cartilage-specific matrix proteins collagen type II, aggrecan, matrix-metalloproteinases (MMP)-3, -13 and pro-inflammatory cytokines were evaluated by real-time RT-PCR and immunohistochemistry. Transduced chondrocytes overexpressed high levels of IL-10 which significantly up-regulated collagen type II expression. TNF-alpha suppressed collagen type II and aggrecan, but increased MMP and cytokine expression in chondrocytes compared to the non-stimulated controls. The TNF-alpha mediated down-regulation of aggrecan expression was significantly antagonized by IL-10 overexpression, whereas the suppression of collagen type II was barely affected. The MMP-13 and IL-1beta expression by TNF-alpha was slightly reduced by IL-10. These results suggest that IL-10 overexpression modulates some catabolic features of TNF-alpha in chondrocytes.
authors.affiliation: Department of Trauma and Reconstructive Surgery, Charité-University of Medicine, Campus Benjamin Franklin, FEM, Krahmerstr. 6-10, 12207 Berlin, Germany.
authors.forename: R D
authors.initials: RD
authors.lastname: Müller
authors.forename: T
authors.initials: T
authors.lastname: John
authors.forename: B
authors.initials: B
authors.lastname: Kohl
authors.forename: A
authors.initials: A
authors.lastname: Oberholzer
authors.forename: T
authors.initials: T
authors.lastname: Gust
authors.forename: A
authors.initials: A
authors.lastname: Hostmann
authors.forename: M
authors.initials: M
authors.lastname: Hellmuth
authors.forename: D
authors.initials: D
authors.lastname: Laface
authors.forename: B
authors.initials: B
authors.lastname: Hutchins
authors.forename: G
authors.initials: G
authors.lastname: Laube
authors.forename: R W
authors.initials: RW
authors.lastname: Veh
authors.forename: S K
authors.initials: SK
authors.lastname: Tschoeke
authors.forename: W
authors.initials: W
authors.lastname: Ertel
authors.forename: G
authors.initials: G
authors.lastname: Schulze-Tanzil
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism; D000368:Aged; D000369:Aged, 80 and over; D001158:Arteries / Q000378:metabolism; D064236:Cartilage Oligomeric Matrix Protein; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects* / Q000378:metabolism* / Q000648:ultrastructure; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D016753:Interleukin-10 / Q000235:genetics / Q000378:metabolism*; D064235:Matrilin Proteins; D053509:Matrix Metalloproteinase 13 / Q000235:genetics / Q000378:metabolism; D019278:Matrix Metalloproteinase 3 / Q000235:genetics / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D008875:Middle Aged; D012333:RNA, Messenger / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha
chemical_list: D064236:Cartilage Oligomeric Matrix Protein; D024043:Collagen Type II; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D064235:Matrilin Proteins; D012333:RNA, Messenger; C577553:TSP5 protein, human; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D053509:Matrix Metalloproteinase 13; D019278:Matrix Metalloproteinase 3