Anti-TNF effects on destructive fibroblasts depend on mechanical stress.
Joint destruction in rheumatoid arthritis (RA) starts typically at sites of mechanically stressed inserts of the synovial membrane near the cartilage/bone border. In the therapy of RA, tumour necrosis factor (TNF) antagonists have rapidly emerged as a valuable class of anti-rheumatic agents that reduce joint destruction. The aim of this study was to investigate and profile genes involved in the interaction between articular movement and anti-TNF therapy in an in vitro model. Murine LS48 cells, an established substitute for invasive RA synovial fibroblasts, were cultured, stretched and/or treated with anti-TNF-alpha antibody for 24 h. RNA was isolated and gene transcript levels were determined using U74Av2 Affymetrix GeneChips to identify transcriptional events. Positive findings were verified by polymerase chain reaction (PCR). We identified 170 differentially regulated genes, including 44 of particular interest. Gene expression fell into different functional groups that can be explained by RA pathogenesis and experimental conditions. For 21 genes of the 44 of particular interest, regulation could be confirmed by real-time PCR. Remarkably, we found structural as well as functional genes differently regulated between stretched cells, anti-TNF-treated cells, and stretched cells treated with anti-TNF antibody. Additionally, we also found a large number of genes that are apparently not related to the experimental conditions. Mechanical exertion modulates gene expression and subsequently cellular response to anti-TNF therapy. Results in exerted cells correspond to current knowledge regarding RA pathogenesis and underline the relevance of our experimental approach. Finally, the central function of the interleukin-18 system in joint destruction could be confirmed by our findings.
Lange
F
F
Institute of Clinical Immunology, University of Leipzig, Leipzig, Germany. franziska.lange@medizin.uni-leipzig.de
Härtl
S
S
Ungethuem
U
U
Kuban
R J
RJ
Hammerschmidt
S
S
Faber
S
S
Morawietz
L
L
Wirtz
H
H
Emmrich
F
F
Krenn
V
V
Sack
U
U
D000818:Animals; D000906:Antibodies / Q000494:pharmacology*; D001169:Arthritis, Experimental / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D005347:Fibroblasts / Q000187:drug effects*; D005786:Gene Expression Regulation; D005787:Gene Frequency; D051379:Mice; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis; D013314:Stress, Mechanical; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*
D000906:Antibodies; D014409:Tumor Necrosis Factor-alpha