Preclinical Development of AS04

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 626)

Abstract

Recent knowledge on vaccine-induced immunity led to the development of vaccine Adjuvant Systems specially designed and adapted to vaccine needs. AS04 is such a tailored Adjuvant System developed by GlaxoSmithKline Biologicals. This chapter focuses on the methods that were used during the preclinical evaluation of AS04. AS04 consists of the combination of aluminum salts and 3-O-deacylated monophosphoryl lipid A (MPL), a detoxified lipid A derivative with retained immunostimulatory capa- city. MPL also induces considerably less pro-inflammatory cytokines, as compared to the parent LPS molecule. Preclinical evaluation of AS04 allowed the determination of the optimal size of MPL particles. The added value of MPL in AS04-based formulations was evidenced by higher vaccine-elicited antibody responses, as well as the induction of higher levels of memory B cells, as compared to aluminum alone formulations. Preclinical evaluation demonstrated the relevance of using AS04 in situations where high and long-lasting antibody levels are needed. This represents the basis for the successful application of AS04 in vaccines against hepatitis B virus and human papillomavirus.

Key words

AS04 Adjuvant System MPL formulation immune response TLR-4 vaccine 

Notes

Acknowledgments

The author is thankful to Ulrike Krause and Pascal Cadot for their assistance in the preparation of the chapter. FENDrix and Cervarix are trade marks of the GlaxoSmithKline group of companies.

References

  1. 1.
    Takeda, K., Akira, S. (2005) Toll-like receptors in innate immunity. Int Immunol 17(1), 1–14.PubMedCrossRefGoogle Scholar
  2. 2.
    Beutler, B., Jiang, Z., Georgel, P., et al. (2006) Genetic analysis of host resistance: Toll-like receptor signaling and immunity at large. Annu Rev Immunol 24, 353–389.PubMedCrossRefGoogle Scholar
  3. 3.
    McKee, A. S., Munks, M. W., Marrack, P. (2007) How do adjuvants work? Important considerations for new generation adjuvants. Immunity 27(5), 687–690.PubMedCrossRefGoogle Scholar
  4. 4.
    Glenny, A. T., Buttlem, G. A. H., Stevens, M. F. (1931) Rate of disappearance of diphtheria toxoid injected into rabbits and guinea pigs: toxoid precipitated with alum. J Pathol 34, 267–275.CrossRefGoogle Scholar
  5. 5.
    Ramon, G. (1926) Procédés pour accroître la production des antitoxines. Ann Inst Pasteur 40, 1–10.Google Scholar
  6. 6.
    Garçon, N., Van Mechelen, M., Wettendorff, M. (2006) Development and evaluation of AS04, a novel and improved immunological adjuvant system containing MPL and aluminium salt, in (Schijns V., O’Hagan D., eds.) Immunopotentiators in Modern Vaccines. Elsevier Academic Press, London, pp 161–177.CrossRefGoogle Scholar
  7. 7.
    Giannini, S. L., Hanon, E., Moris, P., et al. (2006) Enhanced humoral and memory B cellular immunity using HPV16/18 L1 VLP vaccine formulated with the MPL/aluminium salt combination (AS04) compared to aluminium salt only. Vaccine 24(33–34), 5937–5949.PubMedCrossRefGoogle Scholar
  8. 8.
    Myers, K. R., Truchot, A. T., Word, J., Hudson, Y., Ulrich, J. T. (1990) A critical determinant of lipid A endotoxic activity, in (Nowotny A., Spitzer J. J., Ziegler E. J., eds.) Cellular and Molecular Aspects of Endotoxin Reactions. Elsevier Science Publishing Co., New York, pp 145–156.Google Scholar
  9. 9.
    Johnson, D. A., Keegan, D. S., Sowell, C. G., et al. (1999) 3-O-Deacyl monophosphoryl lipid A derivatives: synthesis and immunostimulant activities. J Med Chem 42(22), 4640–4649.PubMedCrossRefGoogle Scholar
  10. 10.
    Hirschfeld, M., Ma, Y., Weis, J. H., Vogel, S. N., Weis, J. J. (2000) Cutting edge: repurification of lipopolysaccharide eliminates signaling through both human and murine Toll-like receptor 2. J Immunol 165(2), 618–622.PubMedGoogle Scholar
  11. 11.
    Tapping, R. I., Akashi, S., Miyake, K., Godowski, P. J., Tobias, P. S. (2000) Toll-like receptor 4, but not Toll-like receptor 2, is a signaling receptor for Escherichia and Salmonella lipopolysaccharides. J Immunol 165(10), 5780–5787.PubMedGoogle Scholar
  12. 12.
    Iwanaga, S., Morita, T., Harada, T., et al. (1978) Chromogenic substrates for horseshoe crab clotting enzyme. Its application for the assay of bacterial endotoxins. Haemostasis 7(2–3), 183–188.PubMedGoogle Scholar
  13. 13.
    Nakamura, S., Morita, T., Iwanaga, S., Niwa, M., Takahashi, K. (1977) A sensitive substrate for the clotting enzyme in horseshoe crab hemocytes. J Biochem 81(5), 1567–1569.PubMedGoogle Scholar
  14. 14.
    European Pharmacopea. (1971). Test for pyrogens, Vol. 11. Published under the direction of the Council of Europe, Maisonneuve SA, Sainte-Ruffine, France, pp 58–60.Google Scholar
  15. 15.
    The European Medicines Agency (EMEA). (2005) Committee for medicinal products for human use (CHMP). Guideline on adjuvants in vaccines for human use. (EMEA/CHMP/VEG/134716/2004) (http://www.emea.europa.eu/pdfs/human/vwp/13471604en.pdf).
  16. 16.
    World Health Organization. (2005) Guidelines on nonclinical evaluation of vaccines. WHO Technical Report Series No. 927, Annex 1, pp 31–63 (http://www.who.int/biologicals/publications/trs/areas/vaccines/nonclinical_evaluation/en/).
  17. 17.
    Bernasconi, N. L., Traggiai, E., Lanzavecchia, A. (2002) Maintenance of serological memory by polyclonal activation of human memory B cells. Science 298(5601), 2199–2202.PubMedCrossRefGoogle Scholar
  18. 18.
    Harper, D. M., Franco, E. L., Wheeler, C., et al. (2004) Efficacy of a bivalent L1 virus-like particle vaccine in prevention of infection with human papillomavirus types 16 and 18 in young women: a randomised controlled trial. Lancet 364(9447), 1757–1765.PubMedCrossRefGoogle Scholar
  19. 19.
    Harper, D., Gall, S., Naud, P., et al. (2008) Sustained immunogenicity and high efficacy against HPV-16/18 related cervical neoplasia: long-term follow up through 6.4 years in women vaccinated with Cervarix™ (GSK’s HPV 16/18 AS04 candidate vaccine). Gynecol Oncol 109(1), 158–159.CrossRefGoogle Scholar
  20. 20.
    Kong, N. C. T., Beran, J., Kee, S. A., et al. (2008) A new adjuvant improves the immune response to hepatitis B vaccine in hemodialysis patients. Kidney Int 73(7), 856–862.PubMedCrossRefGoogle Scholar
  21. 21.
    Beran, J. (2008) Safety and immunogenicity of a new hepatitis B vaccine for the protection of patients with renal insufficiency including pre-haemodialysis and haemodialysis patients. Expert Opin Biol Ther 8(2), 235–247.PubMedCrossRefGoogle Scholar
  22. 22.
    Kool, M., Pétrilli, V., De Smedt, T., Rolaz, A., Hammad, H., van Nimwegen, M., Bergen, I. M., Castillo, R., Lambrecht, B. N., Tschopp, J. (2008) Cutting edge: Alum adjuvant stimulates inflammatory dendritic cells through activation of the NALP3 inflammasome. J Immunol 181(6), 3755–3759, Sep 15.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Global Adjuvant Center for VaccineGlaxoSmithKline BiologicalsWavreBelgium

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