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:

Single dose of a vaccine based on DNA encoding mycobacterial hsp65 protein plus TDM-loaded PLGA microspheres protects mice against a virulent strain of Mycobacterium tuberculosis

Gene Therapy volume 10, pages 678–685 (2003)Cite this article

Abstract

The high incidence of tuberculosis around the world and the inability of BCG to protect certain populations clearly indicate that an improved vaccine against tuberculosis is needed. A single antigen, the mycobacterial heat shock protein hsp65, is sufficient to protect BALB/c mice against challenge infection when administered as DNA vaccine in a three-dose-based schedule. In order to simplify the vaccination schedule, we coencapsulated hsp65-DNA and trehalose dimicolate (TDM) into biodegradable poly(DL-lactide-co-glycolide) (PLGA) microspheres. BALB/c mice immunized with a single dose of DNA-hsp65/TDM-loaded microspheres produced high levels of IgG2a subtype antibody and high amounts of IFN-γ in the supernatant of spleen cell cultures. DNA-hsp65/TDM-loaded microspheres were also able to induce high IFN-γ production in bulk lung cells from challenged mice and confer protection as effective as that attained after three doses of naked DNA administration. This new formulation also allowed a ten-fold reduction in the DNA dose when compared to naked DNA. Thus, this combination of DNA vaccine and adjuvants with immunomodulatory and carrier properties holds the potential for an improved vaccine against tuberculosis.

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

Similar content being viewed by others

References

  1. World Health Organization. Global tuberculosis control. WHO Report 2001. Geneva, Switzerland, WHO/CDS/TB/2001.287.

  2. Ginsberg AM . A proposed national strategy for tuberculosis vaccine development. Clin Infect Dis 2000; 30 (Suppl 3): S233-S242.

    Google Scholar 

  3. Enserink M . Driving a stake into resurgent TB. Science 2001; 293: 234-235.

    Google Scholar 

  4. Thomson J . New TB vaccine to be tested. Nat Med. 2001; 7: 1263.

    Article  CAS  PubMed  Google Scholar 

  5. Silva CL, Lowrie DB . A single mycobacterial protein (hsp65) expressed by a transgenic antigen-presenting cell vaccinates mice against tuberculosis. Immunology 1994; 82: 244–248.

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Lowrie DB, Tascon RE, Colston MJ, Silva CL . Towards a DNA vaccine against tuberculosis. Vaccine 1994; 12: 1537–1540.

    Article  CAS  PubMed  Google Scholar 

  7. Lowrie DB, Silva CL, Colston MJ . Protection against tuberculosis by plasmid DNA. Vaccine 1997; 15: 834–838.

    Article  CAS  PubMed  Google Scholar 

  8. Bonato VLD, Lima VMF, Tascon RE . Identification and characterization of protective T cells in hsp65 DNA vaccinated and Mycobacterium tuberculosis infected mice. Infect Immun 1998; 66: 169–175.

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Lowrie DB, Tascon RE, Bonato VLD . Therapy of tuberculosis in mice by DNA vaccination. Nature 1999; 400: 269–271.

    Article  CAS  PubMed  Google Scholar 

  10. Lima KM, Bonato VLD, Faccioli LH . Comparison of different delivery systems of vaccination for the induction of protection against tuberculosis in mice. Vaccine 2001; 19: 3518–3525.

    Article  CAS  PubMed  Google Scholar 

  11. Gregoriadis G . Genetic vaccines: strategies for optimization. Pharm Res 1998; 15: 661–669.

    Article  CAS  PubMed  Google Scholar 

  12. Lima KM, Santos SA, Santos Jr RR . DNA vaccine encoding mycobacterial hsp65 lacks protectivity when administered by gene gun (submitted).

  13. Wang D, Robinson DR, Kwon GS, Samuel J . Encapsulation of plasmid DNA in biodegradable poly(D,L-lactic-co-glycolic acid) microspheres as a novel approach for immunogene delivery. J Control Rel 1999; 57: 9–18.

    Article  CAS  Google Scholar 

  14. Recent advances on the use of biodegradable microparticles and nanoparticles in controlled drug delivery. Int J Pharm 1995; 116: 1–9.

  15. Eldridge JH, Staas JK, Meulbroek JA . Biodegradable microspheres as a vaccine delivery system. Mol Immun 1991; 28: 287–294.

    Article  CAS  Google Scholar 

  16. Lima KM, Rodrigues Jr JM . Poly-DL-lactide-co-glycolide microspheres as a controlled release antigen delivery system. Braz J Med Biol Res 1999; 32: 171–180.

    Article  CAS  PubMed  Google Scholar 

  17. Krieg AM . From bugs to drugs: therapeutic immunomodulation with oligodeoxynucleotides containing CpG sequences from bacterial DNA. Antisense Nucleic Acid Drug Dev 2001; 11: 181–188.

    Article  CAS  PubMed  Google Scholar 

  18. Bersa GS, Chatterjee D . Lipids and carbohydrates of Mycobacterium tuberculosis. In: Bloom BR (ed). Tuberculosis: Pathogenesis, Protection and Control. ASM Press: Washington, 1994, pp 285–306.

  19. Saito R, Tanaka A, Sugiyama K . Adjuvant effect of cord factor, a mycobacterial lipid. Infect Immun 1976; 13: 766–781.

    Google Scholar 

  20. Koike M, Yoo YC, Mitobe M . Enhancing activity of mycobacterial cell-derived adjuvants on immunogenicity of recombinant human hepatitis B virus vaccine. Vaccine 1998; 16: 1982–1989.

    Article  CAS  PubMed  Google Scholar 

  21. Lima VMF, Bonato VLD, Lima KM . Role of trehalose dimycolate in recruitment of cells and modulation of production of cytokines and NO in tuberculosis. Infect Immun 2001; 69: 5305–5312.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Johansen P, Estevez F, Zurbriggen R . Towards clinical testing of a single-administration tetanus vaccine based on PLA/PLGA microspheres. Vaccine 2000; 19: 1047–1054.

    Article  CAS  PubMed  Google Scholar 

  23. Morein B, Villacrés-Eriksson M, Sjölander A, Bengtsson KL . Novel adjuvants and vaccine delivery systems. Vet Immunol Immunopathol 1996; 54: 373–384.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Klinman DM, Sechler JMG, Conover J . Contribution of cells at the site of DNA vaccination to the generation of antigen-specific immunity and memory. J Immunol 1998; 160: 2388–2392.

    CAS  PubMed  Google Scholar 

  25. Akbari O, Panjwani N, Garcia S . DNA vaccination: transfection and activation of dendritic cells as key events for immunity. J Exp Med 1999; 189: 169–177.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Boyle JS, Brady JL, Lew AM . Enhanced responses to a DNA vaccine encoding a fusion antigen that is directed to sites of immune induction. Nature 1998; 392: 408–411.

    Article  CAS  PubMed  Google Scholar 

  27. Doe B, Selby M, Barnett S . Induction of cytotoxic T lymphocytes by intramuscular immunization with plasmid DNA is facilitated by bone marrow-derived cells. Proc Natl Acad Sci USA 1996; 93: 8578–8583.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Lewis DH . Controlled release of bioactive agents from lactide/glycolide polymers. In: Chasin M, Langer R (eds). Biodegradable Polymers as Drug Delivery Systems. Marcel Dekker: New York, 1990, pp 1–43.

    Google Scholar 

  29. Spargo BJ, Crowe LM, Ioneda T . Cord factor (α,α-trehalose 6-6′-dimycolate) inhibits fusion between phospholipid-vesicles. Proc Natl Acad Sci USA 1991; 88: 737–740.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Ryll R, Watanabe K, Fujiwara N . Mycobacterial cord factor, but not sulfolipid, causes depletion of NKT cells and upregulation of CD1d1 on murine macrophages. Microb Infect 2001; 3: 611–619.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors are grateful to Izaíra T Brandão for technical assistance. This study was supported by grants from Fundac̃ão de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and Conselho Nacional de Pesquisa (CNPq).

Author information

Authors and Affiliations

  1. Department of Biochemistry and Immunology, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil

    K M Lima, S A Santos, A A M Coelho-Castelo, J M Rodrigues Jr & C L Silva

  2. School of Veterinary Medicine, São Paulo State University, Araçatuba, São Paulo, Brazil

    V M F Lima

Authors
  1. K M Lima
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S A Santos
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. V M F Lima
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A A M Coelho-Castelo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J M Rodrigues Jr
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. C L Silva
    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

Lima, K., Santos, S., Lima, V. et al. Single dose of a vaccine based on DNA encoding mycobacterial hsp65 protein plus TDM-loaded PLGA microspheres protects mice against a virulent strain of Mycobacterium tuberculosis. Gene Ther 10, 678–685 (2003). https://doi.org/10.1038/sj.gt.3301908

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301908

Keywords

This article is cited by

Search

Advanced search

Quick links