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:

Recombinant adeno-associated virus-mediated alpha-1 antitrypsin gene therapy prevents type I diabetes in NOD mice

Abstract

Type I diabetes results from an autoimmune destruction of the insulin-producing pancreatic β cells. Although the exact immunologic processes underlying this disease are unclear, increasing evidence suggests that immunosuppressive, immunoregulatory and anti-inflammatory agents can interrupt the progression of the disease. Alpha 1 antitrypsin (AAT) is a multifunctional serine proteinase inhibitor (serpin) that also displays a wide range of anti-inflammatory properties. To test the ability of AAT to modulate the development of type I diabetes, we performed a series of investigations involving recombinant adeno-associated virus vector (rAAV)-mediated gene delivery of human alpha-1 antitrypsin (hAAT) to nonobese diabetic (NOD) mice. Recombinant AAV-expressing hAAT (rAAV2-CB-AT) was administered intramuscularly to 4-week-old female NOD mice (1 × 1010 i.u./mouse). A single injection of this vector reduced the intensity of insulitis, the levels of insulin autoantibodies, and the frequency of overt type I diabetes (30% (3/10) at 32 weeks of age versus 70% (7/10) in controls). Transgene expression at the injection sites was confirmed by immunostaining. Interestingly, antibodies against hAAT were present in a majority of the vector-injected mice and circulating hAAT was undetectable when assessed 10 weeks postinjection. This study suggests a potential therapeutic role for AAT in preventing type I diabetes as well as the ability of AAV gene therapy-based approaches to ameliorate disease effectively.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Atkinson MA, Leiter EH . The NOD mouse model of type 1 diabetes: as good as it gets? Nat Med 1999; 5: 601–604.

    CAS  Google Scholar 

  2. Bendelac A, Carnaud C, Boitard C, Bach JF . Syngeneic transfer of autoimmune diabetes from diabetic NOD mice to healthy neonates. Requirement for both L3T4+ and Lyt-2+ T cells. J Exp Med 1987; 166: 823–832.

    Article  CAS  Google Scholar 

  3. Miller BJ, Appel MC, O'Neil JJ, Wicker LS . Both the Lyt-2+ and L3T4+ T cell subsets are required for the transfer of diabetes in nonobese diabetic mice. J Immunol 1988; 140: 52–58.

    CAS  PubMed  Google Scholar 

  4. Wang Y, Hao L, Gill RG, Lafferty KJ . Autoimmune diabetes in NOD mouse is L3T4 T-lymphocyte dependent. Diabetes 1987; 36: 535–538.

    Article  CAS  Google Scholar 

  5. Like AA et al. Prevention of diabetes in BioBreeding/Worcester rats with monoclonal antibodies that recognize T lymphocytes or natural killer cells. J Exp Med 1986; 164: 1145–1159.

    Article  CAS  Google Scholar 

  6. Sibley RK, Sutherland DE . Pancreas transplantation. An immunohistologic and histopathologic examination of 100 grafts. Am J Pathol 1987; 128: 151–170.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Haskins K et al. T-lymphocyte clone specific for pancreatic islet antigen. Diabetes 1988; 37: 1444–1448.

    Article  CAS  Google Scholar 

  8. Macen JL, Upton C, Nation N, McFadden G . SERP1, a serine proteinase inhibitor encoded by myxoma virus, is a secreted glycoprotein that interferes with inflammation. Virology 1993; 195: 348–363.

    Article  CAS  Google Scholar 

  9. Miyamoto Y et al. Novel functions of human alpha(1)-protease inhibitor after S-nitrosylation: inhibition of cysteine protease and antibacterial activity. Biochem Biophys Res Commun 2000; 267: 918–923.

    Article  CAS  Google Scholar 

  10. Ray MB, Desmet VJ, Gepts W . Alpha-1-Antitrypsin immunoreactivity in islet cells of adult human pancreas. Cell Tissue Res 1977; 185: 63–68.

    Article  CAS  Google Scholar 

  11. Perlmutter DH et al. Expression of the alpha 1-proteinase inhibitor gene in human monocytes and macrophages. Proc Natl Acad Sci USA 1985; 82: 795–799.

    Article  CAS  Google Scholar 

  12. Olsen GN et al. Alpha-1-antitrypsin content in the serum, alveolar macrophages, and alveolar lavage fluid of smoking and nonsmoking normal subjects. J Clin Invest 1975; 55: 427–430.

    Article  CAS  Google Scholar 

  13. Geboes K et al. Morphological identification of alpha-I-anti-trypsin in the human small intestine. Histopathology 1982; 6: 55–60.

    Article  CAS  Google Scholar 

  14. Keppler D et al. Human colon carcinoma cells synthesize and secrete alpha 1-proteinase inhibitor. Biol Chem Hoppe Seyler 1996; 377: 301–311.

    Article  CAS  Google Scholar 

  15. Boskovic G, Twining SS . Local control of alpha1-proteinase inhibitor levels: regulation of alpha1-proteinase inhibitor in the human cornea by growth factors and cytokines. Biochim Biophys Acta 1998; 1403: 37–46.

    Article  CAS  Google Scholar 

  16. Perlmutter DH, Punsal PI . Distinct and additive effects of elastase and endotoxin on expression of alpha 1 proteinase inhibitor in mononuclear phagocytes. J Biol Chem 1988; 263: 16499–16503.

    CAS  PubMed  Google Scholar 

  17. Perlmutter DH, May LT, Sehgal PB . Interferon beta 2/interleukin 6 modulates synthesis of alpha 1-antitrypsin in human mononuclear phagocytes and in human hepatoma cells. J Clin Invest 1989; 84: 138–144.

    Article  CAS  Google Scholar 

  18. Knoell DL, Ralston DR, Coulter KR, Wewers MD . Alpha 1-antitrypsin and protease complexation is induced by lipopolysaccharide, interleukin-1beta, and tumor necrosis factor-alpha in monocytes. Am J Respir Crit Care Med 1998; 157: 246–255.

    Article  CAS  Google Scholar 

  19. Fischer DC et al. Induction of alpha1-antitrypsin synthesis in human articular chondrocytes by interleukin-6-type cytokines: evidence for a local acute-phase response in the joint. Arthritis Rheum 1999; 42: 1936–1945.

    Article  CAS  Google Scholar 

  20. Dhami R et al. Acute cigarette smoke-induced connective tissue breakdown is mediated by neutrophils and prevented by alpha1-antitrypsin. Am J Respir Cell Mol Biol 2000; 22: 244–252.

    Article  CAS  Google Scholar 

  21. Churg A et al. Alpha-1-antitrypsin and a broad spectrum metalloprotease inhibitor, RS113456, have similar acute anti-inflammatory effects. Lab Invest 2001; 81: 1119–1131.

    Article  CAS  Google Scholar 

  22. Waugh JM et al. Therapeutic elastase inhibition by alpha-1-antitrypsin gene transfer limits neointima formation in normal rabbits. J Vasc Interv Radiol 2001; 12: 1203–1209.

    Article  CAS  Google Scholar 

  23. Song S et al. Stable therapeutic serum levels of human alpha-1 antitrypsin (AAT) after portal vein injection of recombinant adeno-associated virus (rAAV) vectors. Gene Therapy 2001; 8: 1299–1306.

    Article  CAS  Google Scholar 

  24. Song S et al. Sustained secretion of human alpha-1-antitrypsin from murine muscle transduced with adeno-associated virus vectors. Proc Natl Acad Sci USA 1998; 95: 14384–14388.

    Article  CAS  Google Scholar 

  25. Goudy K et al. Adeno-associated virus vector-mediated IL-10 gene delivery prevents type 1 diabetes in NOD mice. Proc Natl Acad Sci USA 2001; 98: 13913–13918.

    Article  CAS  Google Scholar 

  26. Sharp HL . The current status of alpha-1-antityrpsin, a protease inhibitor, in gastrointestinal disease. Gastroenterology 1976; 70: 611–621.

    CAS  PubMed  Google Scholar 

  27. Carrell RW et al. Structure and variation of human alpha 1-antitrypsin. Nature 1982; 298: 329–334.

    Article  CAS  Google Scholar 

  28. Perlmutter DH et al. Identification of a serpin-enzyme complex receptor on human hepatoma cells and human monocytes. Proc Natl Acad Sci USA 1990; 87: 3753–3757.

    Article  CAS  Google Scholar 

  29. Zaidi SH et al. Targeted overexpression of elafin protects mice against cardiac dysfunction and mortality following viral myocarditis. J Clin Invest 1999; 103: 1211–1219.

    Article  CAS  Google Scholar 

  30. Niemann MA, Baggott JE, Miller EJ . Binding of SPAAT, the 44-residue C-terminal peptide of alpha 1-antitrypsin, to proteins of the extracellular matrix. J Cell Biochem 1997; 66: 346–357.

    Article  CAS  Google Scholar 

  31. Ziady AG et al. Chain length of the polylysine in receptor-targeted gene transfer complexes affects duration of reporter gene expression both in vitro and in vivo. J Biol Chem 1999; 274: 4908–4916.

    Article  CAS  Google Scholar 

  32. Goudy KS et al. Elucidation of time and dose dependencies using AAV-IL-10 gene therapy for prevention of type 1 diabetes in the NOD mouse. Mol Ther 2002; 5: S17 (abstr. 46).

    Google Scholar 

  33. Zhang YC et al. Genetic predisposition to autoimmunity specifically imparts responsiveness to transgenes delivered by recombinant adeno-associated virus. Mol Ther 2002; 5: S430 (abstr. 1317).

    Google Scholar 

  34. Zhang YC et al. Adeno-associated virus transduction of islets with interleukin-4 results in impaired metabolic function in syngeneic marginal islet mass transplantation. Transplantation 2002; 74(8): 1184–1186.

    Article  CAS  Google Scholar 

  35. Song S, Laipis PJ, Berns KI, Flotte TR . Effect of DNA-dependent protein kinase on the molecular fate of the rAAV2 genome in skeletal muscle. Proc Natl Acad Sci USA 2001; 98: 4084–4088.

    Article  CAS  Google Scholar 

  36. Barbour KW et al. The murine alpha(1)-proteinase inhibitor gene family: polymorphism, chromosomal location, and structure. Genomics 2002; 80: 515–522.

    Article  CAS  Google Scholar 

  37. Joslin G et al. The SEC receptor recognizes a pentapeptide neodomain of alpha 1-antitrypsin-protease complexes. J Biol Chem 1991; 266: 11282–11288.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This study was supported by the grants from the NIH (DK58327, HL59412, RR00082, DK62652), the Juvenile Diabetes Research Foundation, the Alpha 1 Foundation and the Children's Miracle Network.

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Song, S., Goudy, K., Campbell-Thompson, M. et al. Recombinant adeno-associated virus-mediated alpha-1 antitrypsin gene therapy prevents type I diabetes in NOD mice. Gene Ther 11, 181–186 (2004). https://doi.org/10.1038/sj.gt.3302156

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3302156

Keywords

This article is cited by

Search

Quick links