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Differential effects of TNFalpha and IL1beta on FLS global gene expression profile


ABSTRACT: TNFalpha and IL1beta play a pathogenic role in rheumatoid arthritis. Both cytokines are known to activate cytokine and metalloproteinase secretion by synovial fibroblasts. In the present study, we wanted to investigate whether TNFalpha and IL1beta displayed differential effects on cultured Fibroblast-like Synovial Cells derived from RA patients. Global gene expression analyses indicated that both cytokines induced similar genes in these cells. Fibroblast-like cells (FLS) were purified from synovial biopsies from RA patients. Briefly, minced synovial fragments were digested in 1 mg/ml hyaluronidase solution (Sigma Aldrich, St Louis, MO) for 15 minutes at 37M-BM-0C and 6 mg/ml collagenase type IV (Invitrogen, Paisley, UK) for 2 hours at 37M-BM-0C. Next, cells were washed, resuspended in high-glucose DulbeccoM-bM-^@M-^Ys Modified Eagle Medium (Invitrogen) supplemented with 1% antibiotics-antimycotics (Invitrogen) and 1% MEM sodium pyruvate (Invitrogen), and seeded at 10,000 cells/cm2 in 6-well plates. When the cells reached confluence, adherent cells were detached using sterile 0.5% trypsin-EDTA (Invitrogen) and used as FLS between passages 3 and 9. Cells were seeded in 24-well plated at 25.000/well and incubated overnight with or without the following cytokines : TNF-M-NM-1 (R&D Systems, Minneapolis, MN) 10 ng/ml, IL-1M-NM-2 (R&D Systems) 10 ng/ml. After overnight incubation with the indicated cytokines, cells were harvested and total RNA was extracted using the NucleospinM-BM-. RNA II extraction kit (Macherey-Nagel, DM-CM-

ORGANISM(S): Homo sapiens

SUBMITTER: Bernard Lauwerys 

PROVIDER: E-GEOD-15615 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Gene expression profiling in the synovium identifies a predictive signature of absence of response to adalimumab therapy in rheumatoid arthritis.

Badot Valérie V   Galant Christine C   Nzeusseu Toukap Adrien A   Theate Ivan I   Maudoux Anne-Lise AL   Van den Eynde Benoît J BJ   Durez Patrick P   Houssiau Frédéric A FA   Lauwerys Bernard R BR  

Arthritis research & therapy 20090423 2


<h4>Introduction</h4>To identify markers and mechanisms of resistance to adalimumab therapy, we studied global gene expression profiles in synovial tissue specimens obtained from severe rheumatoid arthritis (RA) patients before and after initiation of treatment.<h4>Methods</h4>Paired synovial biopsies were obtained from the affected knee of 25 DMARD (disease-modifying antirheumatic drug)-resistant RA patients at baseline (T0) and 12 weeks (T12) after initiation of adalimumab therapy. DAS28-CRP (  ...[more]

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