Abstract
The enzyme-linked immunospot (ELISPOT) assay is a sensitive and relatively simple assay for detecting secreted cellular products such as cytokines and has become an invaluable immunological tool. The ELISPOT has been used extensively in human and murine research but has only recently been used to assess the feline immune system. For researchers studying feline disease or using the cat as a model of human disease, the quantification of cytokine-producing cells by ELISPOT is an invaluable technique for investigations of disease immunopathogenesis and vaccine efficacy. For example, use of the interferon (IFN)-γ ELISPOT to measure the frequency of antigen-specific T-cells during feline immunodeficiency virus (FTV) infection or after immunization with candidate FTV vaccines is of particular interest. This application of the ELISPOT may serve to expand the utility of FIV as a model for human immunodeficiency virus. Broader applications of the ELISPOT should further our understanding of feline diseases and be useful in the rational development of more efficacious vaccines and therapeutic modalities for the enhancement of feline health. This chapter discusses important parameters of ELISPOT design that will enable researchers to develop and analyze the feline-specific assays within their own laboratory.
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References
Czerkinsky C, Andersson G., Ekre H. P., Nilsson L. A., Klareskog L., and Ouchterlony O. (1988) Reverse ELISPOT assay for clonal analysis of cytokine production. I. Enumeration of gamma-interferon-secreting cells. J. Immunol. Methods 110, 29–36.
Czerkinsky C, Andersson G., Ferrua B., Nordstrom I., Quiding M., Eriksson K., et al. (1991) Detection of human cytokine-secreting cells in distinct anatomical compartments. Immunol. Rev. 119, 5–22.
Czerkinsky C. Nilsson C. L. A., Nygren H., Ouchterlony O., and Tarkowski A. (1983) A solid-phase enzyme-linked immunospot (ELISPOT) assay for enumeration of specific antibody-secreting cells. J. Immunol. Methods 65, 109–121.
Moller S. A. and Borrebaeck C. A. (1985) A filter immuno-plaque assay for the detection of antibody-secreting cells in vitro. J. Immunol. Methods 79, 195–204.
Sedgwick J. D., and Holt P. G. (1983) A solid-phase immunoenzymatic technique for the enumeration of specific antibody-secreting cells. J. Immunol. Methods 57, 301–309.
De Jongh R., Vranken J., Vundelinckx G., Bosmans E., Maes M., and Heylen R. (1997) The effects of anticoagulation and processing on assays of IL-6, sIL-6R, sIL-2R and soluble transform receptor. Cytokine 9, 696–701.
Riches P., Gooding R., Millar B. C, and Rowbottom A. W. (1992) Influence of collection and separation of blood samples on plasma IL-1, IL-6 and TNF-alpha concentrations. J. Immunol. Methods 153, 125–131.
Kreher C. R., Dittrich M. T., Guerkov R., Boehm B. O., and Tary-Lehmann M. (2003) CD4+ and CD8+ cells in cryopreserved human PBMC maintain full functionality in cytokine ELISPOT assays. J. Immunol. Methods 278, 79–93.
Schmittel A., Keilholz U., Bauer S., Kuhne U., Stevanovic S., Thiel E., and Scheibenbogen C (2001) Application of the IFN-gamma ELISPOT assay to quantify T-cell responses against proteins. J. Immunol. Methods 247, 17–24.
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© 2005 Humana Press Inc., Totowa, NJ
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Nordone, S.K., Stevens, R., LaVoy, A.S., Dean, G.A. (2005). Feline Cytokine ELISPOT. In: Kalyuzhny, A.E. (eds) Handbook of ELISPOT. Methods in Molecular Biology™, vol 302. Humana Press. https://doi.org/10.1385/1-59259-903-6:167
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DOI: https://doi.org/10.1385/1-59259-903-6:167
Publisher Name: Humana Press
Print ISBN: 978-1-58829-469-2
Online ISBN: 978-1-59259-903-5
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