title: Lentiviral small hairpin RNA knockdown of macrophage inflammatory protein-1γ ameliorates experimentally induced osteoarthritis in mice.
abstract: Immune cells are involved in the pathogenesis of osteoarthritis (OA). CD4(+) T cells were activated during the onset of OA and induced macrophage inflammatory protein (MIP)-1γ expression and subsequent osteoclast formation. We evaluated the effects of local knockdown of MIP-1γ in a mouse OA model induced by anterior cruciate ligament transection. The mouse macrophage cell lines and osteoclast-like cells generated from immature hematopoietic monocyte/macrophage progenitors of murine bone marrow were cocultured with either receptor activator of NFκB ligand (RANKL) or CD4(+) T cells. The levels of MIP-1γ and RANKL in cells and mice were examined by enzyme-linked immunosorbent assay (ELISA). The osteoclastogenesis was evaluated using tartrate-resistant acid phosphatase and cathepsin K staining. OA was induced in one hind-leg knee joint of B6 mice. Lentiviral vector encoding MIP-1γ small hairpin RNA (shRNA) and control vector were individually injected intra-articularly into the knee joints, which were histologically assessed for manifestations of OA. The expression of MIP-1γ and matrix metalloproteinase (MMP)-13 and the infiltration of CD4(+) T cells, macrophages, and osteoclastogenesis in tissues were examined using immunohistochemistry. CD4(+) T cells were involved in OA by inducing MIP-1γ expression in osteoclast progenitors and the subsequent osteoclast formation. Neutralizing MIP-1γ with a specific antibody abolishes RANKL-stimulated and CD4(+) T-cell-stimulated osteoclast formation. MIP-1γ levels were significantly higher in synovium and the chondro-osseous junction of joints 90 days postsurgery. The number of infiltrated CD4(+) T cells and macrophages and IL-1β expression were reduced in the synovial tissues of mice treated with MIP-1γ shRNA. Histopathological examinations revealed that mice treated with MIP-1γ shRNA had less severe OA than control mice had, as well as decreased osteoclast formation and MMP-13 expression. Locally inhibiting MIP-1γ expression may ameliorate disease progression and provide a new OA therapy.
authors.affiliation: 1 Department of Orthopedic Surgery, Tainan Hospital , Department of Health, Executive Yuan, Tainan 70043, Taiwan .
authors.forename: Po-Chuan
authors.initials: PC
authors.lastname: Shen
authors.forename: Chia-Sing
authors.initials: CS
authors.lastname: Lu
authors.forename: Ai-Li
authors.initials: AL
authors.lastname: Shiau
authors.forename: Che-Hsin
authors.initials: CH
authors.lastname: Lee
authors.forename: I-Ming
authors.initials: IM
authors.lastname: Jou
authors.forename: Jeng-Long
authors.initials: JL
authors.lastname: Hsieh
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002460:Cell Line; D019742:Chemokines, CC / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D004195:Disease Models, Animal; D018450:Disease Progression; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors / Q000235:genetics*; D007719:Knee Joint / Q000378:metabolism / Q000473:pathology; D016086:Lentivirus / Q000235:genetics*; D019402:Macrophage Inflammatory Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D010003:Osteoarthritis / Q000235:genetics* / Q000276:immunology* / Q000473:pathology / Q000628:therapy; D010010:Osteoclasts / Q000187:drug effects / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; C095218:Ccl9 protein, mouse; D019742:Chemokines, CC; D019402:Macrophage Inflammatory Proteins; D034741:RNA, Small Interfering
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