Use of a differential subtraction method to identify genes that characterize the phenotype of cultured rheumatoid arthritis synoviocytes

248

201

Autoimmune collagen-induced arthritis (CIA) in IFN-γR-deficient DBA/1 mice was shown to be reduced in severity by treatment with the bicyclam derivative AMD3100, a specific antagonist of the interaction between the chemokine stromal cell-derived factor-1 (SDF-1) and its receptor CXCR4. The beneficial effect of the CXCR4 antagonist was demonstrable when treatment was initiated between the time of immunization and appearance of the first symptoms. Treatment also reduced the delayed-type hypersensitivity response to the autoantigen, collagen type II. These observations are indicative of an action on a late event in the pathogenesis, such as chemokine-mediated attraction of leukocytes toward joint tissues. The notion of SDF-1 involvement was further supported by the observation that exogenous SDF-1 injected in periarthritic tissue elicited an inflammatory response that could be inhibited by AMD3100. The majority of leukocytes harvested from inflamed joints of mice with CIA were found to be Mac-1+ and CXCR4+, and AMD3100 was demonstrated to interfere specifically with chemotaxis and Ca2+ mobilization induced in vitro by SDF-1 on Mac-1+/CXCR4+ splenocytes. We conclude that SDF-1 plays a central role in the pathogenesis of murine CIA, by attracting Mac-1+/CXCR4+ cells to the inflamed joints.

Section: Discussionsupporting

confidence: 89%

AMD3100, a Potent and Specific Antagonist of the Stromal Cell-Derived Factor-1 Chemokine Receptor CXCR4, Inhibits Autoimmune Joint Inflammation in IFN-γ Receptor-Deficient Mice

Matthys¹,

Hatse

Vermeire³

et al. 2001

248

201

“…In addition, SDF-1 mRNA was expressed by cultured RA synoviocytes. Of note, previous studies indicated that SDF-1 mRNA expression was unique to RA and not OA synoviocytes (48). The current data show that cultured RA fibroblasts produce SDF-1 protein.…”

Section: Cd45rosupporting

confidence: 57%

Stromal Cell-Derived Factor-1-CXC Chemokine Receptor 4 Interactions Play a Central Role in CD4+ T Cell Accumulation in Rheumatoid Arthritis Synovium

Nanki

Hayashida

El-Gabalawy

et al. 2000

427

338

Rheumatoid arthritis (RA) is characterized by the accumulation of CD4+ memory T cells in the inflamed synovium. To address the mechanism, we analyzed chemokine receptor expression and found that the frequency of CXC chemokine receptor (CXCR)4 expressing synovial tissue CD4+ memory T cells was significantly elevated. CXCR4 expression could be enhanced by IL-15, whereas stromal cell-derived factor (SDF)-1, the ligand of CXCR4, was expressed in the RA synovium and could be increased by CD40 stimulation. SDF-1 stimulated migration of rheumatoid synovial T cells and also inhibited activation-induced apoptosis of T cells. These results indicate that SDF-1-CXCR4 interactions play important roles in CD4+ memory T cell accumulation in the RA synovium, and emphasize the role of stromal cells in regulating rheumatoid inflammation.

“…As a way of confirmation, we also analyzed the mRNA expression of genes based on published knowledge from synoviocyte studies. Table I shows genes known to be constitutively overexpressed in RA synoviocytes compared with OA synoviocytes (caldesmon 1 (28), biglycan (28), aggrecan 1 (28), VCAM-1 (29,30), IGFBP-5 (29, 31), CXCL12 (29,31), CD147/basigin (32), MMP-2 (32), and IL-6 (22, 28)), genes known to be expressed at low levels compared with OA synoviocytes (CXCL2 (28) and CXCL6 (28)) and genes known not to be expressed in RA synoviocytes (IL-10, IL-13, TNF-␣, IL-15, and IL-18 (22)). The accuracy of our microarray datasets was further tested through the analysis of expression data upon stimulation (Table I).…”

Section: Transcript Profiling Data Analysis and Validationmentioning

confidence: 99%

Genome-Wide Comparison between IL-17A- and IL-17F-Induced Effects in Human Rheumatoid Arthritis Synoviocytes

Zrioual

Écochard

Tournadre

et al. 2009

157

158

IL-17A is implicated in rheumatoid arthritis (RA) pathogenesis; however, the contribution of IL-17F remains to be clarified. Using microarrays and gene-specific expression assays, we compared the regulatory effects of IL-17A and IL-17F alone or in combination with TNF-α on RA synoviocytes. IL-17A and IL-17F expression was studied in osteoarthritis and RA synovium by immunohistochemistry. The comparison between the IL-17A and IL-17F stimulatory effect on RA synoviocytes was assessed at the protein level by ELISA and at the mRNA level by microarrays and real-time RT-PCR. TNFRII expression was studied by real-time RT-PCR and immunofluorescence, and neutralizing Ab was used to analyze its contribution to CCL20 secretion. IL-17A and IL-17F were detected in plasma cell-like cells from RA but not osteoarthritis synovium. In microarrays, IL-17A and IL-17F alone had similar regulatory effects, IL-17F being quantitatively less active. Both cytokines induced a similar expression pattern in the presence of TNF-α. Based on a cooperation index, 130 and 203 genes were synergistically induced by IL-17A or IL-17F plus TNF-α, respectively. Among these, the new target genes CXCR4, LPL, and IL-32 were validated by real-time RT-PCR. IL-17A and IL-17F up-regulated TNFRII expression, but had no effects on TNFRI, IL-17RA or IL-17RC. TNFRII blockade inhibited the synergistic induction of CCL20 by IL-17A or IL-17F and TNF-α. IL-17A and IL-17F are both expressed in RA synovium. In the presence of TNF-α, they induced a similar expression pattern in RA synoviocytes. Accordingly, IL-17F appears as a target in Th17-mediated diseases such as RA.