Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

An inflammation-inducible adenoviral expression system for local treatment of the arthritic joint

Gene Therapy volume 11, pages 581–590 (2004)Cite this article

Abstract

To achieve a disease-regulated transgene expression for physiologically responsive gene therapy of arthritis, a hybrid promoter was constructed. The human IL-1β enhancer region (−3690 to −2720) upstream of the human IL-6 promoter region (−163 to +12) was essential in mounting a robust response in HIG-82 synovial fibroblasts and in RAW 264,7 macrophages. A replication-deficient adenovirus was engineered with luciferase (Luc) controlled by the IL-1/IL-6 promoter (Ad5.IL-1/IL-6-Luc). LPS caused a 23- and 4.6-fold induction of Luc. activity in RAW cells infected with Ad5.IL-1/IL-6-Luc or the conventional Ad5.CMV-Luc construct, respectively. Next, adenoviruses (106 ffu) were injected into the knees of C57Bl/6 mice. An intra-articular injection of zymosan, 3 days after Ad5.IL-1/IL-6-Luc, increased Luc. activity by 39-fold but had no effect in the Ad5.CMV-Luc joints. The constitutive CMV promoter was rapidly silenced and could not be reactivated in vivo. In contrast, the IL-1/IL-6 promoter could be reactivated by Streptococcal cell wall (SCW)-induced arthritis up to 21 days after infection. Next the IL-1/IL-6 promoter was compared to the C3-Tat/HIV-LTR two-component system in wild-type, IL-6−/− and IL-1−/− gene knockout mice. Both systems responded well to LPS-, zymosan- and SCW-induced arthritis. However, the basal activity of the IL-1/IL-6 promoter was lower and IL-6 independent. This study showed that the IL-1/IL-6 promoter is feasible to achieve disease-regulated transgene expression for treatment of arthritis.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9

Similar content being viewed by others

References

  1. Mattsson R et al. Maintained pregnancy levels of oestrogen afford complete protection from post-partum exacerbation of collagen-induced arthritis. Clin Exp Immunol 1991; 85: 41–47.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Ostensen M, Aune B, Husby G . Effect of pregnancy and hormonal changes on the activity of rheumatoid arthritis. Scand J Rheumatol 1983; 12: 69–72.

    Article  CAS  PubMed  Google Scholar 

  3. Bischof RJ, Zafiropoulos D, Hamilton JA, Campbell IK . Exacerbation of acute inflammatory arthritis by the colony-stimulating factors CSF-1 and granulocyte macrophage (GM)-CSF: evidence of macrophage infiltration and local proliferation. Clin Exp Immunol 2000; 119: 361–367.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Lens JW et al. Flare-up of antigen-induced arthritis in mice after challenge with intravenous antigen: effects of pre-treatment with cobra venom factor and anti-lymphocyte serum. Clin Exp Immunol 1984; 57: 520–528.

    CAS  PubMed  PubMed Central  Google Scholar 

  5. van de Loo AA, Arntz OJ, van den Berg WB . Flare-up of experimental arthritis in mice with murine recombinant IL-1. Clin Exp Immunol 1992; 87: 196–202.

    CAS  PubMed  PubMed Central  Google Scholar 

  6. van Lent PL et al. Phagocytic synovial lining cells regulate acute and chronic joint inflammation after antigenic exacerbation of smouldering experimental murine arthritis. J Rheumatol 1998; 25: 1135–1145.

    CAS  PubMed  Google Scholar 

  7. van de Loo AA et al. Role of interleukin 1 in antigen-induced exacerbations of murine arthritis. Am J Pathol 1995; 146: 239–249.

    CAS  PubMed  PubMed Central  Google Scholar 

  8. van de Loo FA, van den Berg WB . Gene therapy for rheumatoid arthritis. Lessons from animal models, including studies on interleukin-4, interleukin-10, and interleukin-1 receptor antagonist as potential disease modulators. Rheum Dis Clin N Am 2002; 28: 127–149.

    Article  Google Scholar 

  9. Baragi VM . MFG-IRAP University of Pittsburgh. Curr Opin Invest Drugs 2000; 1: 194–198.

    CAS  Google Scholar 

  10. Evans CH et al. Clinical trial to assess the safety, feasibility, and efficacy of transferring a potentially anti-arthritic cytokine gene to human joints with rheumatoid arthritis. Hum Gene Ther 1996; 7: 1261–1280.

    Article  CAS  PubMed  Google Scholar 

  11. Roth S et al. [Anti-TNF alpha monoclonal antibodies (infliximab) and tuberculosis: apropos of 3 cases]. Rev Med Interne 2002; 23: 312–316.

    Article  CAS  PubMed  Google Scholar 

  12. Mayordomo L, Marenco JL, Gomez-Mateos J, Rejon E . Pulmonary miliary tuberculosis in a patient with anti-TNF-alpha treatment. Scand J Rheumatol 2002; 31: 44–45.

    Article  PubMed  Google Scholar 

  13. Nunez MO et al. Reactivation tuberculosis in a patient with anti-TNF-alpha treatment. Am J Gastroenterol 2001; 96: 1665–1666.

    Google Scholar 

  14. Sicotte NL, Voskuhl RR . Onset of multiple sclerosis associated with anti-TNF therapy. Neurology 2001; 57: 1885–1888.

    Article  CAS  PubMed  Google Scholar 

  15. Imhof MO, Chatellard P, Mermod N . Comparative study and identification of potent eukaryotic transcriptional repressors in gene switch systems. J Biotechnol 2002; 97: 275–285.

    Article  CAS  PubMed  Google Scholar 

  16. Varley AW, Geiszler SM, Gaynor RB, Munford RS . A two-component expression system that responds to inflammatory stimuli in vivo. Nat Biotechnol 1997; 15: 1002–1006.

    Article  CAS  PubMed  Google Scholar 

  17. Bakker AC et al. C3-Tat/HIV-regulated intraarticular human interleukin-1 receptor antagonist gene therapy results in efficient inhibition of collagen-induced arthritis superior to cytomegalovirus-regulated expression of the same transgene. Arthritis Rheum 2002; 46: 1661–1670.

    Article  CAS  PubMed  Google Scholar 

  18. Miagkov AV, Varley AW, Munford RS, Makarov SS . Endogenous regulation of a therapeutic transgene restores homeostasis in arthritic joints. J Clin Invest 2002; 109: 1223–1229.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. van de Loo FA et al. Role of interleukin-1, tumor necrosis factor alpha, and interleukin-6 in cartilage proteoglycan metabolism and destruction. Effect of in situ blocking in murine antigen- and zymosan-induced arthritis. Arthritis Rheum 1995; 38: 164–172.

    Article  CAS  PubMed  Google Scholar 

  20. Kuiper S et al. Different roles of tumour necrosis factor alpha and interleukin 1 in murine streptococcal cell wall arthritis. Cytokine 1998; 10: 690–702.

    Article  CAS  PubMed  Google Scholar 

  21. Kim YM, Son K . A nitric oxide production bioassay for interferon-gamma. J Immunol Methods 1996; 198: 203–209.

    Article  CAS  PubMed  Google Scholar 

  22. Kline JN et al. Synergistic activation of the human cytomegalovirus major immediate early promoter by prostaglandin E2 and cytokines. Exp Lung Res 1998; 24: 3–14.

    Article  CAS  PubMed  Google Scholar 

  23. Stein J et al. Tumor necrosis factor alpha stimulates the activity of the human cytomegalovirus major immediate early enhancer/promoter in immature monocytic cells. J Gen Virol 1993; 74: 2333–2338.

    Article  CAS  PubMed  Google Scholar 

  24. Ritter T et al. Stimulatory and inhibitory action of cytokines on the regulation of hCMV-IE promoter activity in human endothelial cells. Cytokine 2000; 12: 1163–1170.

    Article  CAS  PubMed  Google Scholar 

  25. Loser P, Jennings GS, Strauss M, Sandig V . Reactivation of the previously silenced cytomegalovirus major immediate-early promoter in the mouse liver: involvement of NFkappaB. J Virol 1998; 72: 180–190.

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Prosch S et al. Stimulation of the human cytomegalovirus IE enhancer/promoter in HL-60 cells by TNFalpha is mediated via induction of NF-kappaB. Virology 1995; 208: 197–206.

    Article  CAS  PubMed  Google Scholar 

  27. Cheeran MC, Hu S, Gekker G, Lokensgard JR . Decreased cytomegalovirus expression following proinflammatory cytokine treatment in primary human astrocytes. J Immunol 2000; 164: 926–933.

    Article  CAS  PubMed  Google Scholar 

  28. Qin L et al. Promoter attenuation in gene therapy: interferon-gamma and tumor necrosis factor-alpha inhibit transgene expression. Hum Gene Ther 1997; 8: 2019–2029.

    Article  CAS  PubMed  Google Scholar 

  29. Razin A . CpG methylation, chromatin structure and gene silencing – a three-way connection. EMBO J 1998; 17: 4905–4908.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Curradi M, Izzo A, Badaracco G, Landsberger N . Molecular mechanisms of gene silencing mediated by DNA methylation. Mol Cell Biol 2002; 22: 3157–3173.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Prosch S et al. Inactivation of the very strong HCMV immediate early promoter by DNA CpG methylation in vitro. Biol Chem Hoppe Seyler 1996; 377: 195–201.

    Article  CAS  PubMed  Google Scholar 

  32. Benihoud K et al. Efficient, repeated adenovirus-mediated gene transfer in mice lacking both tumor necrosis factor alpha and lymphotoxin alpha. J Virol 1998; 72: 9514–9525.

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Zhang HG et al. Inhibition of tumor necrosis factor alpha decreases inflammation and prolongs adenovirus gene expression in lung and liver. Hum Gene Ther 1998; 9: 1875–1884.

    Article  CAS  PubMed  Google Scholar 

  34. Elkon KB et al. Tumor necrosis factor alpha plays a central role in immune-mediated clearance of adenoviral vectors. Proc Natl Acad Sci USA 1997; 94: 9814–9819.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Peng Y et al. Inhibition of tumor necrosis factor alpha by an adenovirus-encoded soluble fusion protein extends transgene expression in the liver and lung. J Virol 1999; 73: 5098–5109.

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Pan RY et al. Therapy and prevention of arthritis by recombinant adeno-associated virus vector with delivery of interleukin-1 receptor antagonist. Arthritis Rheum 2000; 43: 289–297.

    Article  CAS  PubMed  Google Scholar 

  37. Pan RY et al. Disease-inducible transgene expression from a recombinant adeno-associated virus vector in a rat arthritis model. J Virol 1999; 73: 3410–3417.

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Teschendorf C, Warrington Jr KH, Siemann DW, Muzyczka N . Comparison of the EF-1 alpha and the CMV promoter for engineering stable tumor cell lines using recombinant adeno-associated virus. Anticancer Res 2002; 22: 3325–3330.

    CAS  PubMed  Google Scholar 

  39. Varley AW, Munford RS . Physiologically responsive gene therapy. Mol Med Today 1998; 4: 445–451.

    Article  CAS  PubMed  Google Scholar 

  40. Bednarik DP, Cook JA, Pitha PM . Inactivation of the HIV LTR by DNA CpG methylation: evidence for a role in latency. EMBO J 1990; 9: 1157–1164.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Kopf M et al. Interleukin 6 influences germinal center development and antibody production via a contribution of C3 complement component. J Exp Med 1998; 188: 1895–1906.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Deng C et al. Resistance to experimental autoimmune myasthenia gravis in IL-6-deficient mice is associated with reduced germinal center formation and C3 production. J Immunol 2002; 169: 1077–1083.

    Article  CAS  PubMed  Google Scholar 

  43. Kawamura N, Singer L, Wetsel RA, Colten HR . Cis- and trans-acting elements required for constitutive and cytokine-regulated expression of the mouse complement C3 gene. Biochem J 1992; 283 (Part 3): 705–712.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Wilson DR et al. A 58-base-pair region of the human C3 gene confers synergistic inducibility by interleukin-1 and interleukin-6. Mol Cell Biol 1990; 10: 6181–6191.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Buonaguro L et al. Effects of the human immunodeficiency virus type 1 Tat protein on the expression of inflammatory cytokines. J Virol 1992; 66: 7159–7167.

    CAS  PubMed  PubMed Central  Google Scholar 

  46. Scala G et al. The expression of the interleukin 6 gene is induced by the human immunodeficiency virus 1 TAT protein. J Exp Med 1994; 179: 961–971.

    Article  CAS  PubMed  Google Scholar 

  47. Bakker AC et al. Prevention of murine collagen-induced arthritis in the knee and the ipsilateral paw by local expression of human interleukin-1 receptor antagonist protein in the knee. Arthritis Rheum 1997; 40: 893–900.

    Article  CAS  PubMed  Google Scholar 

  48. Lechman ER et al. Direct adenoviral gene transfer of viral IL-10 to rabbit knees with experimental arthritis ameliorates disease in both injected and contralateral control knees. J Immunol 1999; 163: 2202–2208.

    CAS  PubMed  Google Scholar 

  49. Boyle DL et al. Intra-articular IL-4 gene therapy in arthritis: anti-inflammatory effect and enhanced Th2 activity. Gene Therapy 1999; 6: 1911–1918.

    Article  CAS  PubMed  Google Scholar 

  50. Kim SH et al. Effective treatment of established murine collagen-induced arthritis by systemic administration of dendritic cells geneticaly modified to express IL-4. J Immunol 2001; 166: 3499–3505.

    Article  CAS  PubMed  Google Scholar 

  51. Kim SH et al. Ex vivo gene delivery of IL-1Ra and soluble TNF receptor confers a distal synergistic therapeutic effect in antigen-induced arthritis. Mol Ther 2002; 6: 591–600.

    Article  CAS  PubMed  Google Scholar 

  52. Chan JM et al. Intraarticular gene transfer of TNFR:Fc suppresses experimental arthritis with reduced systemic distribution of the gene product. Mol Ther 2002; 6: 727–736.

    Article  CAS  PubMed  Google Scholar 

  53. Underhill DM, Ozinsky A . Toll-like receptors: key mediators of microbe detection. Curr Opin Immunol 2002; 14: 103–110.

    Article  CAS  PubMed  Google Scholar 

  54. Horai R et al. Production of mice deficient in genes for interleukin (IL)-1α, IL-1β, IL-1α/β, and IL-1 receptor antagonist shows that IL-1β is crucial in turpentine-induced fever development and glucocorticoid secretion. J Exp Med 1998; 187: 1463–1475.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Kopf M et al. Impaired immune and acute-phase responses in interleukin-6-deficient mice. Nature 1994; 368: 339–342.

    Article  CAS  PubMed  Google Scholar 

  56. Kinoshita S, Akira S, Kishimoto T . A member of the C/EBP family, NF-IL6 beta, forms a heterodimer and transcriptionally synergizes with NF-IL6. Proc Natl Acad Sci USA 1992; 89: 1473–1476.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Abe M et al. Regulation of interleukin (IL)-1beta gene transcription induced by IL-1beta in rheumatoid synovial fibroblast-like cells, E11, transformed with simian virus 40 large T antigen. J Rheumatol 1997; 24: 420–429.

    CAS  PubMed  Google Scholar 

  58. Chartier C et al. Efficient generation of recombinant adenovirus vectors by homologous recombination in Escherichia coli. J Virol 1996; 70: 4805–4810.

    CAS  PubMed  PubMed Central  Google Scholar 

  59. Mittereder N, March KL, Trapnell BC . Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy. J Virol 1996; 70: 7498–7509.

    CAS  PubMed  PubMed Central  Google Scholar 

  60. Fallaux FJ et al. Characterization of 911: a new helper cell line for the titration and propagation of early region 1-deleted adenoviral vectors. Hum Gene Ther 1996; 7: 215–222.

    Article  CAS  PubMed  Google Scholar 

  61. Erbacher P . Methods for adenovirus-mediated gene transfer to airway epithelium. In: Robbins PD (ed). Methods in Molecular Medicine, Gene Therapy Protocols. Humana press Inc.: Totowa, NJ, 1997, pp 169–184.

    Google Scholar 

  62. van Beuningen HM et al. Phenotypic differences in murine chondrocyte cell lines derived from mature articular cartilage. Osteoarthritis Cartilage 2002; 10: 977–986.

    Article  CAS  PubMed  Google Scholar 

  63. Ebbinghaus C et al. Functional and selective targeting of adenovirus to high-affinity Fcgamma receptor I-positive cells by using a bispecific hybrid adapter. J Virol 2001; 75: 480–489.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. van den Broek MF, van den Berg WB, van de Putte LB, Severijnen AJ . Streptococcal cell wall-induced arthritis and flare-up reaction in mice induced by homologous or heterologous cell walls. Am J Pathol 1988; 133: 139–149.

    CAS  PubMed  PubMed Central  Google Scholar 

  65. Butel JS, TAlas M, Ugur J, Melnick JL . Demonstration of infectious DNA in transformed cells. III. Correlation of detection of infectious DNA–protein complexes with persistence of virus in simian adenovirus SA7-induced tumor cells. Intervirology 1975; 5: 43–56.

    Article  CAS  PubMed  Google Scholar 

  66. Morris DJ, Cooper RJ, Barr T, Bailey AS . Polymerase chain reaction for rapid diagnosis of respiratory adenovirus infection. J Infect 1996; 32: 113–117.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank M Netea (Internal Medicine, UMCN, Nijmegen, The Netherlands) for the collaboration on the IL-1α/β gene knockout mice of which the breeding pairs were kindly provided by Y Iwakura (Center for Experimental Medicine, Institute of Medical Science, University of Tokyo, Japan). We also thank M Kopf (Molecular Biomedicine, Department of Environmental Sciences, Swiss Federal Institute of Technology, Zurich, Switzerland) for providing us with the IL-6 gene knockout mice breeding pairs. Furthermore, we express our gratitude to T Kishimoto (Department of Molecular Medicine, Osaka University Graduate School of Medicine, Osaka, Japan) for kindly providing us the human IL-6 promoter vector. The CARex-Fc fusion protein for optimal adenoviral infection of macrophages was kindly provided by S Hemmi (Institute of Molecular Biology, University of Zürich, Zürich, Switzerland).

This research was supported by grants from the Dutch Arthritis Association (941,304) and the Dutch Organization for Scientific Research (902-27-218).

Author information

Authors and Affiliations

  1. Rheumatology Research and Advanced Therapeutic, University Medical Center Nijmegen, Nijmegen, The Netherlands

    FAJ van de Loo, ASK de Hooge, RL Smeets, AC Bakker, MB Bennink, OJ Arntz, LAB Joosten, HM van Beuningen, PK van der Kraan & WB van den Berg

  2. Division of Infectious Diseases, Department of Internal Medicine, Molecular Host Defense Lab, University of Texas Southwestern Medical Center at Dallas, Texas, USA

    AW Varley

Authors
  1. FAJ van de Loo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. ASK de Hooge
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. RL Smeets
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. AC Bakker
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. MB Bennink
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. OJ Arntz
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. LAB Joosten
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. HM van Beuningen
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. PK van der Kraan
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. AW Varley
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. WB van den Berg
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

van de Loo, F., de Hooge, A., Smeets, R. et al. An inflammation-inducible adenoviral expression system for local treatment of the arthritic joint. Gene Ther 11, 581–590 (2004). https://doi.org/10.1038/sj.gt.3302182

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3302182

Keywords

This article is cited by

Search

Advanced search

Quick links