Impact Analysis And Mechanism Investigation To Rat's Adjuvant Arthritis Model Using Anti-DR5 MAb

Rheumatoid arthritis(RA)is an autoimmune disease characterised by chronic inflammation of the joints.Although the precise pathogenesis of RA remains unclear, T cells,B cells,macrophages,neutrophils and synovial fibroblasts are central to the mechanisms of joint inflammation and disease progression.RA is connected with the apoptosis inhibition of synovioblasts and lymphocytes in synovial tissues.To induce apoptosis of these cells could prevent them from excessive proliferation,and therefore would be an effectual way to treat RA.TRAIL(TNF related apoptosis inducing ligand)are typeⅡtransmembrane protein which are members of TNF superfamily.TRAIL can induce apoptosis of those cells expressing receptors that are specific for TRAIL.DR5(also called TRAIL-R2)is the specific receptor to TRAIL,which was highly expressed in many kinds of tumor tissues.To administrate TRAIL could,however,bring about severe hepatotoxicity,thus limite its usage.Through the method of immunizing the mice by soluble DR5(sDR5),our laboratory had established a hybridoma cell line secreting Anti-DR5 monoclonal antibody which can induce some tumor cell to apoptosis.This mAb binds DR5 specifically,and don't react with other TRAIL receptor.Furthermore,the mAb is able to induce apoptosis of some tumor cells which express DR5.In order to investigate the effect and immune mechanism that Anti-DR5 mAb make on the rat's adjuvant arthritis model,I adopted measures as follow:First,we established rat's adjuvant arthritis animal model by injecting complete Freud's adjuvant(CFA)into the joints of rats;After that,I administrated Anti-DR5 mAb to those artificial patients using caudal intravenous injection for the sake of finding out the variation of rat's articular tumidity and the inflammatory cytokines such as IL-1β,TNF-α,IFN-γin serum and synovia.In addition,I used RT-PCR to detect DR5 mRNA expression in each main tissues or organs,synovium and lymphocytes,while used western blot to analyse the expression of apoptosis-related factors that include Caspase-8 and Bcl-2 in the main tissues and organs.During the research process,I was observing the phenomenon that Anti-DR5 mAb brought about the remission of RA and the inprovement of articular swelling.After treated with Anti-DR5 mAb,the rats produced far more less IL-1β,TNF-α,IFN-γin their blood and synovial tissue,and the overmultiplication of synoviocytes and lymphocytes was even restrained.In conclution,Anti-DR5 mAb produced by our lab are capable of treating the rat's AA and acquired well therapeutic effect.The mechanism based on this therapy may be really linked to the expression level of cell surface DR5,cytokines(IL-1β,TNF-α,IFN-γ),Caspase-8 protein.