| BackgroundRheumatoid arthritis (RA) is a systemic autoimmune disease, characterized by joint inflammation and synoviocyte proliferation, always leads to joint destruction eventually. Chemikines are a series of inducible secretary cytokines playing an important role in local infiltration of inflammatory cells in RA and closely associated with synovitis, tissue damage, neovascularization. All of these biological effects are performed by binding of ligands to receptors. Elsewhere, CXCR3 and CCR5 are mark chemokine receptors in RA.IL-8 is an important ligand of CXCR3, with potency activity in chemotaxis of neutrophiles and neovascularization. IP-10, another ligand of CXCR3, is secreted by monocytes, fibroblasts and endothelial cells under the stimulation of IFN-γ, playing an important role in immigration of T cells to inflammatory sites. In contrast to IL-8, IP-10 is an antagonistto neovascularization. RANTES attracts CD4+/CD45RO+ T lymphocyte to immigrate to inflammatory sites. Whatsmore, the expression of RANTES in joints is correlated to the degree of joint destruction.Therefore, we measured serum levels of CXCR3 ligands (IL-8, IP-10) and CCR5 ligand (RANTES) in patients of active RA patients, relieved RA patients, OA patients and healthy control cases, respectively. Their clinical data were compared to explore the role of chemokine in the pathogenesis and progression of RA, and to investigate the relation between serum level of chemokine and common clinical indices of RA, to evaluate the possibility of these chemokine used clinically as a referrible index to assess the activity degrees and prognosis in patient with RA.Objects and Methods ObjectsWe selected continuously 87 patients of rheumatoid arthritis in outpatients and inpatients of the Department of Rheumatism and Immunology in the Second Affiliated Hospital of Zhejiang University Medical School, during November 2002 to March 2003. According to respective inclusive and exclusive standards, they were divided into four groups: 58 patients in active rheumatoid arthritis group(group)which were subdivided into group I a (37 patients with duration after onset no less than 24 months)and group I b (21 patients with duration after onset less than 24 months), 29 patients in relieved RA group (II group), 21 patients in OA group (III group). 30 patients in Healthy control group (IVgroup). Experimental Methods 1. Main reagents1. 1 IL-8 EASIA kit and RANTES EASIA kit purchased from BIOSOURCE EUROPE S.A1. 2 Human IP-10 ELISA kit purchased from CYTIMMUNE SCIENCES INC.2. Peripheral blood collection and serum separation: 5ml fast blood were collected from elbow vein in the morning and put in sterile tube without anticoagulant. Serum samples were separated by centrifuging 3000 rpm for 10 minutes and stored in refrigerate at -80.3. Peripheral serum levels of IL-8, IP-10 and RANTES were measured by a double antibody sandwich enzyme immunoassay method according to the protocol. Read the microtiter plate at 450nm. Construct a standard curve using all standard points for which absorbance is below the limit of reader used. Determine IL-8, IP-10 or RANTES concentrations of samples or controls individually according to absorbance reading by the standard curve. Multiple by the dilution factor if necessary in order to obtain the concentration in undiluted sample.Measurement of the biochemical indices4. ESR (Erythrocyte sedimentation rate) were measured by-Rheumatoidfactor (RF), C reaction protein (CRP), complements ( C3 , C4 ) and immunoglobulin (IgG IgM IgA) were measured by Immune rate nephelometry. Unit of ESR is mm/h. Uite of RF is IU/ml. Unit of CRP is mg/L. Unit of IgG IgM, IgA is g/L. Unit of C3 is g/L. Unit of C4 is mg/L.5. Review of X-ray image of hand and wrist joints of active RA patients: According to severity of joint changes, the radiological image of hand and wrist joints of RA patients is divided into five grades from I to IV, and considered as 1 to 4 points accordingly.6. Statistical analysis: All data were adm...
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