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Volume 3 Supplement 2

21st European Workshop for Rheumatology Research

  • Meeting abstract
  • Open Access

Development of a doxycycline inducible AAV vector for long term in vivo viral IL-10 gene transfer in rheumatoid arthritis

  • 1,
  • 1,
  • 1,
  • 1,
  • 1, 2 and
  • 1, 2
Arthritis Research & Therapy20013 (Suppl 2) :P091

  • Received: 15 January 2001
  • Published:


  • Doxycycline
  • DBA1 Mouse
  • Rheumatoid Synovial Tissue
  • Inject Control Group
  • Cytokine Gene Transfer


The recent development of AAV vectors (adeno-associated virus) offers new perspectives for cytokine gene transfer in RA as they are non pathogenic and allow long term transgene expression in vivo. Moreover, we propose to regulate vIL-10 expression with tetracycline derivative (tetON system). The purpose of this study was to assess the potential long-term gene expression regulation of a recombinant AAV vector expressing vIL-10 in human rheumatoid synovial tissue and its efficiency in collagen-induced arthritis (CIA).


The AAV-tetON-vIL10 vector contains two transcriptional units oriented in opposite directions, with a central bi-directional SV40 polyA. Sequences encoding the transcriptional activator rtTA, which confers doxycycline transgene induction, is inserted downstream a retroviral LTR promoter. A minimal human CMV promoter, flanked with tetracycline operator motifs, controls the transcription of vIL-10. Human RA synoviocytes were infected in vitro with AAV-tetON-vIL10 (500 MOI) and vIL-10 secretion was assessed by ELISA after addition of 5 μg/ml doxycycline (dox). Therapeutic efficiency of the vector was achieved after intra-muscular injection (1.5 × 109 pi) in DBA1 mice with CIA in the presence of doxycycline in the drinking water (0.2 mg/ml).


Viral IL-10 secretion by RA synoviocytes was increased 40-fold in presence of dox (233 ng/ml/106 cells) and returned to basal level 24 hr after dox removal. In CIA, serum vIL-10 increased to 0.38 ng/ml, 5 weeks after gene transfer in animals under diet dox. RT-PCR analysis showed vIL-10 transcription in the muscle up to 14 weeks, without diffusion in other organs. We observed a decrease of CIA incidence (30% versus 89% in AAV-GFP injected control group) and of paw swelling (1.68 ± 0.04 versus 1.81 ± 0.15 on day 35 post-immunization, P < 0.0003).


AAV vectors conferred safe and inducible long-term expression of vIL-10. These data support AAV-tetON-vIL10 as a therapeutical tool for gene therapy in RA.

Authors’ Affiliations

INSERM U475, France
Immunorhumatologie, CHU Lapeyronie, Montpellier, France


© BioMed Central Ltd 2001