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MMP-3 expression and release by rheumatoid arthritis fibroblast-like synoviocytes induced with a bacterial ligand of integrin alpha5beta1.

Zeisel MB, Druet VA, Wachsmann D, Sibilia J - Arthritis Res. Ther. (2004)

Bottom Line: RT-PCR and pro-MMP-3 (pro-matrix metalloproteinase) assays were then performed to confirm the up-regulation of gene expression.Only one gene, that for insulin-like growth factor binding protein-4, was down-regulated in all RA FLSs.In contrast, in osteoarthritis FLSs only one gene, that for IL-6, was modulated.

View Article: PubMed Central - HTML - PubMed

Affiliation: Inserm 392, Infection et Inflammation, Université Louis Pasteur de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67400 Illkirch, France. zeisel@pharma.u-strasbg.fr

ABSTRACT
Fibroblast-like synoviocytes (FLSs) play a major role in the pathogenesis of rheumatoid arthritis (RA) by secreting effector molecules that promote inflammation and joint destruction. How these cells become and remain activated is still elusive. Both genetic and environmental factors probably play a role in transforming FLSs into inflammatory matrix-degrading cells. As bacterial products have been detected in the joint and shown to trigger joint inflammation, this study was undertaken to investigate whether a bacterial ligand of integrin alpha5beta1, protein I/II, could contribute to the aggressive behavior of RA FLSs. Protein I/II is a pathogen-associated molecular pattern (PAMP) isolated from oral streptococci that have been identified in the joints of RA patients. The response of RA and osteoarthritis FLSs to protein I/II was analyzed using human cancer cDNA expression arrays. RT-PCR and pro-MMP-3 (pro-matrix metalloproteinase) assays were then performed to confirm the up-regulation of gene expression. Protein I/II modulated about 6% of all profiled genes. Three of these, those encoding IL-6, leukemia inhibitory factor, and MMP-3, showed a high expression level in all RA FLSs tested, whereas the expression of genes encoding other members of the cytokine or MMP-family was not affected. Furthermore, the up-regulation of MMP-3 gene expression was followed by an increase of pro-MMP-3 release. The expression of interferon regulatory factor 1 and fibroblast growth factor-5 was also up-regulated, although the expression levels were lower. Only one gene, that for insulin-like growth factor binding protein-4, was down-regulated in all RA FLSs. In contrast, in osteoarthritis FLSs only one gene, that for IL-6, was modulated. These results suggest that a bacterial ligand of integrin alpha5beta1 may contribute to the aggressive behavior of RA FLSs by inducing the release of pro-inflammatory cytokines and a cartilage-degrading enzyme, such as IL-6 and MMP-3, respectively.

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Gene expression patterns in rheumatoid arthritis (RA) and osteoarthritis (OA) fibroblast-like synoviocytes (FLSs). Total RNA of control RA (a) and OA FLSs (c) and of RA (b) and OA FLSs (d) stimulated for 4 hours with protein I/II were first reverse-transcribed and 32P-labeled and then hybridized to Atlas human cancer cDNA expression arrays. Arrows indicate double spots representing IL-6.
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Figure 1: Gene expression patterns in rheumatoid arthritis (RA) and osteoarthritis (OA) fibroblast-like synoviocytes (FLSs). Total RNA of control RA (a) and OA FLSs (c) and of RA (b) and OA FLSs (d) stimulated for 4 hours with protein I/II were first reverse-transcribed and 32P-labeled and then hybridized to Atlas human cancer cDNA expression arrays. Arrows indicate double spots representing IL-6.

Mentions: In this study we used a gene array technique to further investigate the cellular response of FLSs to protein I/II. As we were particularly interested in genes that could contribute to the aggressive behavior of RA FLSs, we chose Atlas human cancer cDNA expression arrays containing genes involved in cell-cycle regulation, cellular adhesion, and inflammation. In parallel to RA FLSs, we used OA FLSs in order to study the protein-I/II-induced response of FLSs isolated from a noninflammatory arthropathy. After a 4-hour incubation in the presence or absence of 125 pm protein I/II, the total RNA from FLSs of three RA and three OA patients was extracted, and radiolabeled cDNA probes were subsequently hybridized to the cDNA arrays. Figure 1 shows the expression patterns of FLSs isolated from one RA and one OA patient in nonstimulated controls after 4 hours (Fig. 1a,1c) and after stimulation with protein I/II for 4 hours (Fig. 1b,1d). The products of some differentially expressed genes (e.g. the gene for IL-6) are easily observed. The relative expression level of each gene in protein-I/II-stimulated FLSs versus control FLSs was evaluated with the AtlasImage 2.0 Software.


MMP-3 expression and release by rheumatoid arthritis fibroblast-like synoviocytes induced with a bacterial ligand of integrin alpha5beta1.

Zeisel MB, Druet VA, Wachsmann D, Sibilia J - Arthritis Res. Ther. (2004)

Gene expression patterns in rheumatoid arthritis (RA) and osteoarthritis (OA) fibroblast-like synoviocytes (FLSs). Total RNA of control RA (a) and OA FLSs (c) and of RA (b) and OA FLSs (d) stimulated for 4 hours with protein I/II were first reverse-transcribed and 32P-labeled and then hybridized to Atlas human cancer cDNA expression arrays. Arrows indicate double spots representing IL-6.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC1064889&req=5

Figure 1: Gene expression patterns in rheumatoid arthritis (RA) and osteoarthritis (OA) fibroblast-like synoviocytes (FLSs). Total RNA of control RA (a) and OA FLSs (c) and of RA (b) and OA FLSs (d) stimulated for 4 hours with protein I/II were first reverse-transcribed and 32P-labeled and then hybridized to Atlas human cancer cDNA expression arrays. Arrows indicate double spots representing IL-6.
Mentions: In this study we used a gene array technique to further investigate the cellular response of FLSs to protein I/II. As we were particularly interested in genes that could contribute to the aggressive behavior of RA FLSs, we chose Atlas human cancer cDNA expression arrays containing genes involved in cell-cycle regulation, cellular adhesion, and inflammation. In parallel to RA FLSs, we used OA FLSs in order to study the protein-I/II-induced response of FLSs isolated from a noninflammatory arthropathy. After a 4-hour incubation in the presence or absence of 125 pm protein I/II, the total RNA from FLSs of three RA and three OA patients was extracted, and radiolabeled cDNA probes were subsequently hybridized to the cDNA arrays. Figure 1 shows the expression patterns of FLSs isolated from one RA and one OA patient in nonstimulated controls after 4 hours (Fig. 1a,1c) and after stimulation with protein I/II for 4 hours (Fig. 1b,1d). The products of some differentially expressed genes (e.g. the gene for IL-6) are easily observed. The relative expression level of each gene in protein-I/II-stimulated FLSs versus control FLSs was evaluated with the AtlasImage 2.0 Software.

Bottom Line: RT-PCR and pro-MMP-3 (pro-matrix metalloproteinase) assays were then performed to confirm the up-regulation of gene expression.Only one gene, that for insulin-like growth factor binding protein-4, was down-regulated in all RA FLSs.In contrast, in osteoarthritis FLSs only one gene, that for IL-6, was modulated.

View Article: PubMed Central - HTML - PubMed

Affiliation: Inserm 392, Infection et Inflammation, Université Louis Pasteur de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67400 Illkirch, France. zeisel@pharma.u-strasbg.fr

ABSTRACT
Fibroblast-like synoviocytes (FLSs) play a major role in the pathogenesis of rheumatoid arthritis (RA) by secreting effector molecules that promote inflammation and joint destruction. How these cells become and remain activated is still elusive. Both genetic and environmental factors probably play a role in transforming FLSs into inflammatory matrix-degrading cells. As bacterial products have been detected in the joint and shown to trigger joint inflammation, this study was undertaken to investigate whether a bacterial ligand of integrin alpha5beta1, protein I/II, could contribute to the aggressive behavior of RA FLSs. Protein I/II is a pathogen-associated molecular pattern (PAMP) isolated from oral streptococci that have been identified in the joints of RA patients. The response of RA and osteoarthritis FLSs to protein I/II was analyzed using human cancer cDNA expression arrays. RT-PCR and pro-MMP-3 (pro-matrix metalloproteinase) assays were then performed to confirm the up-regulation of gene expression. Protein I/II modulated about 6% of all profiled genes. Three of these, those encoding IL-6, leukemia inhibitory factor, and MMP-3, showed a high expression level in all RA FLSs tested, whereas the expression of genes encoding other members of the cytokine or MMP-family was not affected. Furthermore, the up-regulation of MMP-3 gene expression was followed by an increase of pro-MMP-3 release. The expression of interferon regulatory factor 1 and fibroblast growth factor-5 was also up-regulated, although the expression levels were lower. Only one gene, that for insulin-like growth factor binding protein-4, was down-regulated in all RA FLSs. In contrast, in osteoarthritis FLSs only one gene, that for IL-6, was modulated. These results suggest that a bacterial ligand of integrin alpha5beta1 may contribute to the aggressive behavior of RA FLSs by inducing the release of pro-inflammatory cytokines and a cartilage-degrading enzyme, such as IL-6 and MMP-3, respectively.

Show MeSH
Related in: MedlinePlus