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:

Gene transfer of Fc-fusion cytokine by in vivo electroporation: application to gene therapy for viral myocarditis

Gene Therapy volume 9, pages 577–583 (2002)Cite this article

Abstract

Among a number of techniques for gene transfer in vivo, the direct injection of plasmid DNA into muscle is simple, inexpensive and safe. Although combining direct DNA injection with in vivo electroporation increases the efficiency of gene transfer into muscle, applications of this method have remained limited because of the relatively low expression level. To overcome this problem, we developed a plasmid vector that expresses a secretory protein as a fusion protein with the noncytolytic immunoglobulin Fc portion and used it for electroporation-mediated viral interleukin 10 (vIL-10) expression in vivo. The fusion cytokine vIL-10/mutFc was successfully expressed and the peak serum concentration of vIL-10 was almost 100-fold (195 ng/ml) higher than with a non-fusion vIL-10 expression plasmid. The expressed fusion cytokine suppressed the phytohemagglutinin-induced IFN-γ production by human peripheral blood mononuclear cells and decreased the mortality in a mouse viral myocarditis model as effectively as vIL-10 expression. These results demonstrate that the transfer of plasmid DNA expressing a noncytolytic Fc-fusion cytokine is useful to deliver enhanced levels of cytokine without altering general biological activities. This simple and efficient system should provide a new approach to gene therapy for human diseases and prove very useful for investigating the function of newly discovered secretory protein genes.

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

Similar content being viewed by others

References

  1. Donohue JH, Rosenberg SA . The fate of interleukin-2 after in vivo administration J Immunol 1983 130: 2203–2208

    CAS  PubMed  Google Scholar 

  2. Marchant A et al. Interleukin-10 controls interferon-gamma and tumor necrosis factor production during experimental endotoxemia Eur J Immunol 1994 24: 1167–1171

    Article  CAS  Google Scholar 

  3. Nitta Y et al. Systemic delivery of interleukin 10 by intramuscular injection of expression plasmid DNA prevents autoimmune diabetes in nonobese diabetic mice Hum Gene Ther 1998 9: 1701–1707

    Article  CAS  Google Scholar 

  4. O'Hara RM Jr, Henderson SL, Nagelin A . Prevention of a Th1 disease by a Th1 cytokine: IL-12 and diabetes in NOD mice Ann NY Acad Sci 1996 795: 241–249

    Article  CAS  Google Scholar 

  5. Trembleau S et al. Interleukin 12 administration induces T helper type 1 cells and accelerates autoimmune diabetes in NOD mice J Exp Med 1995 181: 817–821

    Article  CAS  Google Scholar 

  6. Prud'homme GJ, Lawson BR, Chang Y, Theofilopoulos AN . Immunotherapeutic gene transfer into muscle Trends Immunol 2001 22: 149–155

    Article  CAS  Google Scholar 

  7. Aihara H, Miyazaki J . Gene transfer into muscle by electroporation in vivo Nature Biotechnol 1998 16: 867–870

    Article  CAS  Google Scholar 

  8. Nakano A et al. Cytokine gene therapy for myocarditis by in vivo electroporation Hum Gene Ther 2001 12: 1289–1297

    Article  CAS  Google Scholar 

  9. Chernoff AE et al. A randomized, controlled trial of IL-10 in humans. Inhibition of inflammatory cytokine production and immune responses J Immunol 1995 154: 5492–5499

    CAS  Google Scholar 

  10. Lotze MT et al. In vivo administration of purified human interleukin 2. II. Half life, immunologic effects, and expansion of peripheral lymphoid cells in vivo with recombinant IL 2 J Immunol 1985 135: 2865–2875

    CAS  PubMed  Google Scholar 

  11. Wolff JA et al. Direct gene transfer into mouse muscle in vivo Science 1990 247: 1465–1468

    Article  CAS  Google Scholar 

  12. Tokui M et al. Intramuscular injection of expression plasmid DNA is an effective means of long-term systemic delivery of interleukin-5 Biochem Biophys Res Commun 1997 233: 527–531

    Article  CAS  Google Scholar 

  13. Zheng XX et al. A noncytolytic IL-10/Fc fusion protein prevents diabetes, blocks autoimmunity, and promotes suppressor phenomena in NOD mice J Immunol 1997 158: 4507–4513

    CAS  PubMed  Google Scholar 

  14. Crowe PD et al. A lymphotoxin-beta-specific receptor Science 1994 264: 707–710

    Article  CAS  Google Scholar 

  15. Shao H et al. cDNA cloning and characterization of a Cek7 receptor protein-tyrosine kinase ligand that is identical to the ligand (ELF-1) for the Mek-4 and Sek receptor protein-tyrosine kinases J Biol Chem 1995 270: 3467–3470

    Article  CAS  Google Scholar 

  16. Sun QH et al. Individually distinct Ig homology domains in PECAM-1 regulate homophilic binding and modulate receptor affinity J Biol Chem 1996 271: 11090–11098

    Article  CAS  Google Scholar 

  17. Nadeau RW et al. A comparison of mass balance, pharmacokinetics and disposition of [14C(U)]- and [125I]recombinant human interleukin-2 in cynomolgus monkeys Drug Metab Dispos Biol Fate Chem 1995 23: 904–909

    CAS  PubMed  Google Scholar 

  18. Zheng XX et al. Administration of noncytolytic IL-10/Fc in murine models of lipopolysaccharide-induced septic shock and allogeneic islet transplantation J Immunol 1995 154: 5590–5600

    CAS  Google Scholar 

  19. Steurer W et al. Ex vivo coating of islet cell allografts with murine CTLA4/Fc promotes graft tolerance J Immunol 1995 155: 1165–1174

    CAS  Google Scholar 

  20. Duncan AR, Winter G . The binding site for C1q on IgG Nature 1988 332: 738–740

    Article  CAS  Google Scholar 

  21. Hsu DH et al. Expression of interleukin-10 activity by Epstein-Barr virus protein BCRF1 Science 1990 250: 830–832

    Article  CAS  Google Scholar 

  22. Moore KW et al. Homology of cytokine synthesis inhibitory factor (IL-10) to the Epstein–Barr virus gene BCRFI Science 1990 248: 1230–1234

    Article  CAS  Google Scholar 

  23. Vieira P et al. Isolation and expression of human cytokine synthesis inhibitory factor cDNA clones: homology to Epstein–Barr virus open reading frame BCRFI Proc Natl Acad Sci USA 1991 88: 1172–1176

    Article  CAS  Google Scholar 

  24. de Waal Malefyt R . Interleukin 10 (IL-10) and viral IL-10 strongly reduce antigen-specific human T cell proliferation by diminishing the antigen-presenting capacity of monocytes via downregulation of class II major histocompatibility complex expression J Exp Med 1991 174: 915–924

    Article  CAS  Google Scholar 

  25. Matsumori A . Molecular and immune mechanisms in the pathogenesis of cardiomyopathy - role of viruses, cytokines, and nitric oxide Jpn Circ J 1997 61: 275–291

    Article  CAS  Google Scholar 

  26. Ho SN et al. Site-directed mutagenesis by overlap extension using the polymerase chain reaction Gene 1989 77: 51–59

    Article  CAS  Google Scholar 

  27. Niwa H, Yamamura K, Miyazaki J . Efficient selection for high-expression transfectants with a novel eukaryotic vector Gene 1991 108: 193–199

    Article  CAS  Google Scholar 

  28. Suzuki T et al. Viral interleukin 10 (IL-10), the human herpes virus 4 cellular IL-10 homologue, induces local anergy to allogeneic and syngeneic tumors J Exp Med 1995 182: 477–486

    Article  CAS  Google Scholar 

  29. Radloff R, Bauer W, Vinograd J . A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells Proc Natl Acad Sci USA 1967 57: 1514–1521

    Article  CAS  Google Scholar 

  30. Miyazaki J et al. Expression vector system based on the chicken beta-actin promoter directs efficient production of interleukin-5 Gene 1989 79: 269–277

    Article  CAS  Google Scholar 

  31. Abrams JS et al. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples Immunol Rev 1992 127: 5–24

    Article  CAS  Google Scholar 

  32. Overbergh L, Valckx D, Waer M, Mathieu C . Quantification of murine cytokine mRNAs using real time quantitative reverse transcriptase PCR Cytokine 1999 11: 305–312

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr Satwant Narula at Schering-Plough Research Institute for the generous gift of pcDSRα-BCRF. This work was supported by a grant from the Research for the Future Program (JSPS-RFTF97I00201) of the Japan Society for the Promotion of Science (JSPS). This work was also supported by a grant from the Ministry of Education, Science, Sports and Culture.

Author information

Authors and Affiliations

  1. Division of Stem Cell Regulation Research, Osaka University Graduate School of Medicine, Osaka, Japan

    O Adachi, A Nakano, O Sato, E Yamato & J Miyazaki

  2. Department of Cardiovascular Surgery, Tohoku University Graduate School of Medicine, Sendai, Japan

    O Adachi, S Kawamoto & K Tabayashi

  3. Department of Cardiovascular Medicine, Kyoto University Graduate School of Medicine, Kyoto, Japan

    A Nakano & A Matsumori

  4. Department of Surgery, The Institute of Medical Science, University of Tokyo, Tokyo, Japan

    H Tahara

  5. Department of Obstetrics and Gynecology, Mie University School of Medicine, Tsu, Japan

    N Toyoda

Authors
  1. O Adachi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A Nakano
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. O Sato
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S Kawamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H Tahara
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. N Toyoda
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. E Yamato
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. A Matsumori
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. K Tabayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. J Miyazaki
    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

Adachi, O., Nakano, A., Sato, O. et al. Gene transfer of Fc-fusion cytokine by in vivo electroporation: application to gene therapy for viral myocarditis. Gene Ther 9, 577–583 (2002). https://doi.org/10.1038/sj.gt.3301691

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301691

Keywords

This article is cited by

Search

Advanced search

Quick links