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Real-time quantitative PCR to detect changes in synovial gene expression in rheumatoid arthritis after corticosteroid treatment
  1. Danielle M Gerlag1,
  2. David L Boyle2,
  3. Sanna Rosengren2,
  4. Tony Nash3,
  5. Paul P Tak1,
  6. Gary S Firestein2
  1. 1Division of Clinical Immunology and Rheumatology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
  2. 2Division of Rheumatology, Allergy and Immunology, UCSD School of Medicine, La Jolla, California, USA
  3. 3Astra Zeneca, Macclesfield, UK
  1. Correspondence to:
    Professor P P Tak
    Division of Clinical Immunology and Rheumatology, F4-218, Academic Medical Center, University of Amsterdam, PO Box 22700, 1100 DE, Amsterdam, The Netherlands; p.p.tak{at}amc.uva.nl

Abstract

Synovial biomarkers are increasingly important in the development of novel therapeutic agents for the treatment of rheumatoid arthritis (RA). To identify biomarkers correlating with changes in clinical disease activity, real-time quantitative PCR (Q-PCR) was used to evaluate changes in synovial gene expression after treatment with corticosteroids. Patients with active RA received either oral prednisolone (n = 10, 60 mg daily for the first week and 40 mg daily for the second week) or placebo (n = 11) for 14 days. Real-time Q-PCR was used to quantify gene expression of tumour necrosis factor (TNF)α, IL1β, IL8 and matrix metalloproteinase (MMP) 1 in synovial tissue samples obtained through an arthroscopic procedure before and after treatment. mRNA levels were reported as relative expression units compared with a cell-based standard. Statistical analysis was performed using an analysis of covariance model. Prednisolone markedly decreased IL8 and MMP1 expression compared with placebo, and the CIs excluded the likelihood of no effect. A trend towards reduction was seen in IL1β and TNFα mRNA expression in the prednisolone group, although CIs included the value for no effect. These data suggest that Q-PCR can be used to measure synovial mRNA expression of mediators implicated in the pathogenesis of RA in small proof-of-concept trials.

  • c-DNA, complementary DNA
  • CE, cell equivalent
  • C(t), threshold cycle number
  • DAS, disease activity score
  • MMP, matrix metalloproteinase
  • PBMC, peripheral blood monocyte
  • Q-PCR, quantitative PCR
  • RA, rheumatoid arthritis
  • REU, relative expression units
  • TNF, tumour necrosis factor

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Footnotes

  • Published Online First 19 September 2006

  • Competing interests: TN worked for Astra Zeneca when the study was performed, and owns Astra Zeneca stock.

  • The clinical trial was sponsored by Astra Zeneca. DMG was sponsored by the Dutch Arthritis Association.

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