Abstract
CMV-specific T cells may be detected and quantified after antigen-specific stimulation based on the induction of cytokines as a readout system. Secreted cytokines may be detected from the supernatant of stimulated cells using ELISA. Alternatively, antigen-specific cytokine-secreting cells may be enumerated using an ELISPOT assay. These assays generally rely on the detection of IFNγ and do not allow for a simultaneous assessment of several cytokines on a single cell basis. Here we describe a flow-cytometry based method to analyze CMV-specific CD4 T cells after specific stimulation with a whole antigen lysate. In this assay, cytokine secretion from stimulated cells is blocked by the addition of brefeldin A. Using a panel of fluorescently labeled antibodies, not only intracellularly accumulated cytokines but also phenotypical characteristics of specifically activated T cells may be quantified in a multiparameter staining approach.
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References
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Schmidt, T., Sester, M. (2013). Detection of Antigen-Specific T Cells Based on Intracellular Cytokine Staining Using Flow-Cytometry. In: Bailer, S., Lieber, D. (eds) Virus-Host Interactions. Methods in Molecular Biology, vol 1064. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-601-6_19
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DOI: https://doi.org/10.1007/978-1-62703-601-6_19
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-600-9
Online ISBN: 978-1-62703-601-6
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