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AB0076 Analysis for The Gene Expression Profiles in Rheumatoid Synovial Fibroblasts Regulated by Light
  1. T. Maeda1,
  2. Y. Miura1,
  3. K. Fukuda2,
  4. S. Hayashi1,
  5. M. Kurosaka1
  1. 1Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, Kobe
  2. 2Department of Orthopaedic Surgery, Kakogawa City Hospital, Kakogawa, Japan

Abstract

Background Rheumatoid Arthritis (RA) is an autoimmune disease characterized by over-proliferation of synovial tissues and subsequent joint destruction [1]. Lymphotoxin-like, exhibits inducible expression, and competes with herpes simplex virus glycoprotein D for herpes virus entry mediator, a receptor expressed on T lymphocytes (LIGHT) is a member of tumor necrosis factor (TNF) receptor superfamily and is expressed on antigen-presenting cells through the activation of T cells. LIGHT modulates T-cell activation and promotes inflammation through the activation of nuclear factor (NF)κB by binding to its specific receptors herpes virus entry mediator (HVEM) and lymphotoxin (LT)-βR [2]. Furthermore, decoy receptor 3 (DcR3) competitively binds soluble LIGHT and inhibits LIGHT signaling via HVEM [3,4]. Studies have suggested that LIGHT is overexpressed in RA fibroblast-like synoviocytes (RA-FLS) and production of TNFα is induced by stimulation with LIGHT, and that LIGHT may be related to autoimmune diseases such as inflammatory bowel disease or multiple sclerosis [5].

Objectives In this study, we investigated the genes regulated by LIGHT in RA-FLS by comprehensive genetic analysis using microarrays to elucidate the involvement of LIGHT in RA pathogenesis.

Methods Microarray assay. Four individual lines of primary cultured RA-FLS were incubated with either 1.0 μg/ml recombinant human LIGHT protein or phosphate buffered saline (PBS) diluted with serum-free Opti-MEM medium as non-stimulated control for 12 hours at 37°C with 5% CO2. Gene expression was detected using cDNA microarray assay (Human Genome U133 Plus 2.0, GeneChip® 3' Expression Array, Affymetrix), and the gene expression profiles of LIGHT-stimulated cells and controls were compared.

Real-time polymerase chain reaction (real-time PCR). The four most highly up- and down-regulated genes were discovered by comprehensive genetic analysis using the microarray. The relative mRNA expression levels of those genes were compared using TaqMan® real-time PCR on a StepOne™ real-time PCR system.

Results Microarray assay. The microarray analysis revealed that 1042 genes were up-regulated and 801 genes were down-regulated more than twofold by stimulation with LIGHT in RA-FLS.

Expression of LIGHT-related genes in RA-FLS. The real-time PCR analysis confirmed that mRNA expression of the top-four genes up-regulated and those down-regulated was actually regulated by LIGHT.

Conclusions To our knowledge, this is the first report on the expression profiles of genes regulated by LIGHT in RA-FLS, which suggested the involvement of LIGHT-HVEM/DcR3 signaling in the pathogenesis of RA.

  1. Alamanos Y, et al. Autoimmun Rev. 2005 Mar;4:130–6.

  2. Murphy TL, Murphy KM. Annu Rev Immunol. 2010;28:389–411.

  3. Ware CF. Immunol Rev. 2008 Jun;223:186–201.

  4. Wang Y, et al. Immunol Rev. 2009 May;229(1):232–43.

  5. Steinberg MW, et al. Immunol Rev. 2011 Nov;244(1):169–87.

Disclosure of Interest None declared

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