Large-Animal Model for Establishing E/T Ratio of Adjuvants

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

Abstract

To develop novel adjuvants for use in humans, the efficacy/toxicity (E/T) ratio of experimental products in large animal species can be investigated. The test model included two intramuscular immunizations in pigs at 3 weeks interval and analysis of immune responses and local reactions 1 week after the second injection. The antigen used to determine adjuvant activity was a well-defined, purified, viral glycoprotein that without adjuvant induces low immune responses and no detectable local reactions. Efficacy was determined by measuring ELISA and virus-neutralizing antibody titres. Toxicity was determined by necropsy and estimating size and severity of local reactions to each treatment. The persistence of the side effects was deduced from the difference in the local reaction 4 weeks after the first and 1 week after the second injection. For graphic representation of E/T ratios, toxicity was expressed in arbitrary units and plotted against antibody titre. The graphs provided insight into dose– and structure–response relationships and enabled the stepwise optimization of adjuvant candidates.

Key words

Animal model pig efficacy local toxicity reactogenicity E/T ratio necropsy risk/benefit ratio 

References

  1. 1.
    Mestas, J., Hughes, C. W. (2004) Of mice and not men: differences between mouse and human immunology. J Immunol 172, 2731–2738.PubMedGoogle Scholar
  2. 2.
    Hulst, M. M., Westra, D. F., Wensvoort, G., Moormann, R. J. M. (1993) Glycoprotein E1 of hog cholera virus expressed in insect cells protects swine from hog cholera. J Virol 67, 6479–6486.Google Scholar
  3. 3.
    Bokhout, B. A., Bianchi, A. T., van der Heijden, P. J., Scholten, J. W., Stok, W. (1986) The influence of a water-in-oil emulsion on humoral immunity. Comp Immunol Microbiol Infect Dis 9, 161–168.PubMedCrossRefGoogle Scholar
  4. 4.
    Hilgers, L. A., Lejeune, G., Nicolas, I., Fochesato, M., Boon, B. (1999) Sulfolipo-cyclodextrin in squalane-in-water as a novel and safe vaccine adjuvant. Vaccine 17, 219–228.PubMedCrossRefGoogle Scholar
  5. 5.
    Blom, A. G., Hilgers, L. A. T. (2004) Sucrose fatty acid sulphate esters as novel vaccine adjuvants: effect of the chemical composition. Vaccine 23, 743–754.PubMedCrossRefGoogle Scholar
  6. 6.
    Hilgers, L. A. T., Platenburg, P. L. I., Luitjens, A., Groenveld, B., Dazelle, T., Ferrari-Laloux, M., Weststrate, M. W. (1994) A novel non-mineral oil-based adjuvant. I. Efficacy of a synthetic sulfolipopolysaccharide in squalane-in-water emulsion in laboratory animals. Vaccine 12, 653–660.PubMedCrossRefGoogle Scholar
  7. 7.
    Hilgers, L. A. T., Platenburg, P. L. I., Luitjens, A., Groenveld, B., Dazelle, T., Weststrate, M. W. (1994) A novel non-mineral oil-based adjuvant. II. Efficacy of a synthetic sulfolipopolysaccharide in squalane-in-water emulsion in pigs. Vaccine 12, 661–665.PubMedCrossRefGoogle Scholar
  8. 8.
    Bouma, A., de Smit, A. J., de Kluiver, E. P., Terpstra, C., Moormann, R. J. M. (1999) Efficacy and stability of a subunit vaccine based on glycoprotein E2 of classical swine fever virus. Vet Microbiol 66, 101–114.PubMedCrossRefGoogle Scholar
  9. 9.
    Terpstra, C., Bloemraad, M., Gielkens, A. L. J. (1984) The neutralizing peroxidase-linked assay for detection of antibody against swine fever. Vet Microbial 16, 123–128.CrossRefGoogle Scholar
  10. 10.
    US Department of Health and Human Services, Food and Drug Administration, Center for Biologics Evaluation and Research. (2009). Guidance for Industry: toxicity grading scale for healthy adult and adolescent volunteers enrolled in preventive vaccine clinical trials (Draft guidance 2005). Available at http://www.fda.gov/cber/gdlns/toxvac.htm [accessed January 2009].

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Nobilon International BVBoxmeerthe Netherlands

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