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Suppression of collagen-induced arthritis by intra-articular lentiviral vector-mediated delivery of Toll-like receptor 7 short hairpin RNA gene

Gene Therapy volume 19, pages 752–760 (2012)Cite this article

Abstract

Knockdown of Toll-like receptors (TLRs) is a novel therapeutic strategy in treating patients with rheumatoid arthritis (RA). We examined the effects of lentiviral vector-mediated delivery of TLR7 short hairpin RNA gene (Lt.shTLR7) on collagen-induced arthritis (CIA). After being immunized on days 0 and 7, Sprague–Dawley rats received intra-articular (i.a.) injection of Lt.shTLR7 or scramble control vector on days 7 and 10. The therapeutic effects were evaluated by measuring ankle circumferences, articular index, and radiographic and histological scores on killing on day 16. Microvessel densities, vascular endothelial growth factor (VEGF) levels, pro-inflammatory cytokine concentrations and T-cell numbers within the synovial tissues were measured. Moreover, VEGF and pro-inflammatory cytokine concentrations in culture supernatants from TLR7-transfected synovial fibroblasts (SFs) stimulated with imiquimod or endogenous ligands were examined. There were significant reduction in ankle circumferences, articular indexes, and radiographic and histological scores. Microvessel densities, VEGF concentrations, interleukin (IL)-1β and IL-6 levels and T-cell densities within synovial tissues were significantly lower. Induction of VEGF, IL-1β and IL-6 production from stimulated SFs was significantly suppressed. Taken together, these data demonstrate the effects of i.a. lentiviral vector-mediated delivery of shTLR7 RNA gene on inhibition of CIA, and implicate the manipulation of TLR7 as a potential therapeutic strategy in RA patients.

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Acknowledgements

We thank Dr D Trono (Ecole Polytechnique Federale de Lausanne, Lausanne, Switzerland) for kindly providing pLVTHM, psPAX2 and pMD2G plasmids for lentivirus production, and Dr LJ Chang (Department of Molecular Genetics and Microbiology, University of Florida) for providing TE671 cells. This work was supported by the National Science Council (NSC Grants 97-2314-B-006-007-MY3 and 100-2314-B-006-031-MY3), the Department of Health, Executive Yuan (Grants DOH 96-TD-I-111-TM014 and 97-TD-I-111-TM015) and the National Cheng Kung University Landmark Project (Grant NCKU-97-R010), Taiwan, Republic of China.

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  1. S-Y Chen and A-L Shiau: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, National Cheng Kung University, Tainan, Taiwan, Republic of China

    S-Y Chen, Y-T Li & C-L Wu

  2. Department of Internal Medicine, Section of Rheumatology and Immunology, National Cheng Kung University, Tainan, Taiwan, Republic of China

    S-Y Chen & C-R Wang

  3. Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, Tainan, Taiwan, Republic of China

    A-L Shiau, Y-S Lin & C-R Wang

  4. Department of Microbiology, College of Medicine, China Medical University, Taichung, Taiwan, Republic of China

    C-H Lee

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Correspondence to C-L Wu or C-R Wang.

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Chen, SY., Shiau, AL., Li, YT. et al. Suppression of collagen-induced arthritis by intra-articular lentiviral vector-mediated delivery of Toll-like receptor 7 short hairpin RNA gene. Gene Ther 19, 752–760 (2012). https://doi.org/10.1038/gt.2011.173

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