Abstract
Large numbers and functionally competent T cells are required to protect from diseases for which antibody-based vaccines have consistently failed (1), which is the case for many chronic viral infections and solid tumors. Therefore, therapeutic vaccines aim at the induction of strong antigen-specific T-cell responses. Novel adjuvants have considerably improved the capacity of synthetic vaccines to activate T cells, but more research is necessary to identify optimal compositions of potent vaccine formulations. Consequently, there is a great need to develop accurate methods for the efficient identification of antigen-specific T cells and the assessment of their functional characteristics directly ex vivo. In this regard, hundreds of clinical vaccination trials have been implemented during the last 15 years, and monitoring techniques become more and more standardized.
*These authors contributed equally to this work.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Woodland, D. L. (2004) Jump-starting the immune system: prime-boosting comes of age. Trends Immunol 25, 98–104.
Mahnke, Y. D., Speiser, D., Luescher, I. F., et al. (2005) Recent advances in tumour antigen-specific therapy: in vivo veritas. Int J Cancer 113, 173–178.
Andersen, M. H., Sorensen, R. B., Schrama, D., et al. (2008) Cancer treatment: the combination of vaccination with other therapies. Cancer Immunol Immunother 57, 1735–1743.
Romero, P., Cerottini, J. C., Speiser, D. E. (2006) The human T cell response to melanoma antigens. Adv Immunol 92, 187–224.
Vujanovic, L., Butterfield, L. H. (2007) Melanoma cancer vaccines and anti-tumor T cell responses. J Cell Biochem 102, 301–310.
Altman, J. D., Moss, P. A., Goulder, P. J., et al. (1996) Phenotypic analysis of antigen-specific T lymphocytes. Science 274, 94–96.
Czerkinsky, C. C., Nilsson, L. A., Nygren, H., et al. (1983) A solid-phase enzyme-linked immunospot (ELISPOT) assay for enumeration of specific antibody-secreting cells. J Immunol Methods 65, 109–121.
Jung, T., Schauer, U., Heusser, C., et al. (1993) Detection of intracellular cytokines by flow cytometry. J Immunol Methods 159, 197–207.
Baumgaertner, P., Rufer, N., Devevre, E., et al. (2006) Ex vivo detectable human CD8 T-cell responses to cancer-testis antigens. Cancer Res 66, 1912–1916.
Romero, P., Dunbar, P. R., Valmori, D., et al. (1998) Ex vivo staining of metastatic lymph nodes by class I major histocompatibility complex tetramers reveals high numbers of antigen-experienced tumor-specific cytolytic T lymphocytes. J Exp Med 188, 1641–1650.
Speiser, D. E., Pittet, M. J., Guillaume, P., et al. (2004) Ex vivo analysis of human antigen-specific CD8+ T-cell responses: quality assessment of fluorescent HLA-A2 multimer and interferon-gamma ELISPOT assays for patient immune monitoring. J Immunother 27, 298–308.
Pittet, M. J., Valmori, D., Dunbar, P. R., et al. (1999) High frequencies of naive Melan-A/MART-1-specific CD8(+) T cells in a large proportion of human histocompatibility leukocyte antigen (HLA)-A2 individuals. J Exp Med 190, 705–715.
Taswell, C. (1981) Limiting dilution assays for the determination of immunocompetent cell frequencies. I. Data analysis. J Immunol 126, 1614–1619.
Whelan, J. A., Dunbar, P. R., Price, D. A., et al. (1999) Specificity of CTL interactions with peptide-MHC class I tetrameric complexes is temperature dependent. J Immunol 163, 4342–4348.
Appay, V., Nixon, D. F., Donahoe, S. M., et al. (2000) HIV-specific CD8(+) T cells produce antiviral cytokines but are impaired in cytolytic function. J Exp Med 192, 63–75.
Appay, V., Rowland-Jones, S. L. (2002) The assessment of antigen-specific CD8+ T cells through the combination of MHC class I tetramer and intracellular staining. J Immunol Methods 268, 9–19.
Betts, M. R., Brenchley, J. M., Price, D. A., et al. (2003) Sensitive and viable identification of antigen-specific CD8+ T cells by a flow cytometric assay for degranulation. J Immunol Methods 281, 65–78.
Fukuda, M. (1991) Lysosomal membrane glycoproteins. Structure, biosynthesis, and intracellular trafficking. J Biol Chem 266, 21327–21330.
Lyons, A. B., Parish, C. R. (1994) Determination of lymphocyte division by flow cytometry. J Immunol Methods 171, 131–137.
Parish, C. R. (1999) Fluorescent dyes for lymphocyte migration and proliferation studies. Immunol Cell Biol 77, 499–508.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Derré*, L. et al. (2010). Quantitative Multiparameter Assays to Measure the Effect of Adjuvants on Human Antigen-Specific CD8 T-Cell Responses. In: Davies, G. (eds) Vaccine Adjuvants. Methods in Molecular Biology, vol 626. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-585-9_16
Download citation
DOI: https://doi.org/10.1007/978-1-60761-585-9_16
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-584-2
Online ISBN: 978-1-60761-585-9
eBook Packages: Springer Protocols