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| Expression and Biological Characterization of Human Osteoprotegerin Fused with Mycobacteria Heat Shock Protein 65 |
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Abstract A fused functional gene of human OPG and Mhsp65 was amplified by PCR, and cloned into the prokaryotic expression vector pET-28a. The BL21 (DE3) strain of E.coli was transformed using the recombinant plasmid pET-28a-OPG-HSP65 and the expected protein was expressed by induction with IPTG. Result of SDS-PAGE indicated that the expected recombinant protein of 23 kDa was expressed with high yield as inclusion body. The fusion protein could be specifically recognized by both the anti-His antibody and anti-human OPG monoclonal antibody in Western blot analysis. The purified and refolded fusion protein could inhibit osteoclast proliferation and bone absorption in vitro. The results of mouse ear swelling assay and expressions of TNF-α, IFN-γ and IL-17 mRNAs detected by real-time quantitative PCR demonstrated that the fusion protein had an anti-inflammation activity. The results suggest that the fusion protein of human OPG and Mhsp65 may act as a potential therapeutics for rheumatoid arthritis.
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Received: 04 January 2009
Published: 27 April 2009
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| ［1］ 王霖，王文杰.破骨细胞在类风湿关节炎骨破坏中的作用及其调控机制.生理科学进展，2004，35（3）：269~272
Wang L，Wang W J.Progress in Physiological Sciences，2004，35（3）：269~272
［2］ Tanaka S.RANKL as a target molecule for treatment of joint destruction. Clin Calcium，2007，17(4)：586~592
［3］ Kmiec Z，Sokolowska I.Role of tumor necrosis factor family ligands in the pathogenesis of rheumatoid arthritis——new therapeutical opportunities. Pol Merkur Lekarski,2007，22(130)：300~304
［4］ Walsh M C，Choi Y.Biology of the TRANCE axis. Cytokine Growth Factor Rev，2003，14(3-4)：251~263
［5］ Nanki T.Molecular mechanisms of bone destruction in rheumatoid arthritis. Clin Calcium，2007，17(4)：510~516
［6］ Gonalves E D，Bonato V L，da Fonseca，D M，et al.Improve protective efficacy of a TBDNA HSP65 vaccine by BCG priming. Genet Vaccines Ther，2007，22：7~13
［7］ 何文晓，李蕴，赵文明,等.结核杆菌热休克蛋白肽段对胶原诱导性关节炎保护作用的研究.中国免疫学杂志，2006，22(12)：1092~1095
He W X，Li Y，Zhao W M，et al．Chinese Journal of Immunology，2006，22(12)：1092~1095
［8］ 马静，王炜，赵文明,等.人骨保护素分枝杆菌热休克蛋白70融合蛋白的克隆与表达及其活性鉴定.中国组织工程研究与临床康复，2008，12(15)：2903~2909
Ma J，Wang W，Zhao W M，et al.Journal of Clinical Rehabilitative Tissue Engineering Research，2008，12(15)：2903~2909
［9］ Elefteriou F,Ahn J D,Takeda S,et al.Leptin regulation of bone resorption by the sympathetic nervous system and CART. Nature，2005，434(7032)：514~520
［10］ Glass D A,Bialek P,Ahn J D,et al.Canonical Wnt signaling in differentiated osteoblasts controls osteoclast differentiation. Dev Cell，2005，8(5)：751~764
［11］ Yasuda H.OPG，anti-RANKL antibody. Nippon Rinsho，2005，63：1647~1653
［12］ Anandarajah A P，Schwarz E M .Anti-RANKL therapy for inflammatory bone disorders: Mechanisms and potential clinical applications. J Cell Biochem，2006，97:226~232
［13］ van Eden W，van der Zee R，Prakken B.Heat-shock proteins induce T-cell regulation of chronic inflammation. Nat Rev Immunol，2005，5(4)：318~330 |
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