Further, blocking VEGF function by administration of a neutralising VEGF antibody or a soluble VEGF receptor to mice with established CIA, sup pressed disease progression, whereas exogenous VEGF exacerbated synovial inflammation and joint destruction. Based on these observations, sellekchem we utilised the mouse CIA model to investigate the functional capillary density in the inflamed synovium and to analyse genes that are involved in synovial angiogenesis during arthritis. Gene expression profiling of murine CIA has been conducted before. However, these reports were based on a whole genome approach, and many angiogenesis related genes, such as VEGF and its receptors, have not been assessed.

Given that relatively little information is avail able on the kinetics of angiogenesis relevant gene expression in CIA, the present study applied quantitative reverse transcription PCR to investigate the expression of selected genes Inhibitors,Modulators,Libraries at onset, peak and declining phases of arthritis in DBA 1 mice. The hypoxic micro environment in RA, coupled with the paradoxical fea ture of increased synovial vascularity, led us to speculate that the function of the synovial vasculature is disturbed and that abnormal angiogenic activity might contribute to the progression of CIA. We thus hypothesised that angiogenesis related genes, including growth factors and receptors, would correlate with arthritis progression and angiogenic activity, particularly in the established phases of arthritis, allowing us to identify potential molecular targets for therapeutic intervention.

Materials and methods Mice Male DBA 1J and female C57BL 6 mice were pur chased from Harlan Laboratories and maintained under standard conditions at the Biological Services Unit of the Kennedy Institute of Rheumatology, Imperial College London, UK. Studies were performed in accordance with the UK Animals Act 1986 regulations Inhibitors,Modulators,Libraries for the handling and use of laboratory animals, and followed an Ethics Committee and Home Office approved project licence. For intravital microscopy studies, male DBA 1J mice were obtained from Taconic and kept under standard con ditions at the animal care facility of the University of Rostock, Germany. Induction and evaluation of arthritis Type II collagen was purified from bovine articular Inhibitors,Modulators,Libraries cartilage following the method described by Miller. Prior to usage, CII was dissolved in 0. 05M acetic acid.

Complete Freunds Inhibitors,Modulators,Libraries adjuvant was prepared by grinding 100 Inhibitors,Modulators,Libraries mg Mycobacterium tuberculosis and suspending it in 30 ml incomplete Freunds adjuvant. For the induction of CIA, CII was emulsified in equal volumes of CFA to prepare a 1 mg ml solution, and male DBA 1J mice aged 10 12 weeks were injected intradermally more info at the base of the tail with 100 ul. Mice were given a booster injection 21 days post primary immunisation with 50 ��g CII emulsified in IFA. A control group of mice received injections of CFA alone followed by IFA alone 21 days later.