Chapter title |
Four Color ImmunoSpot ® Assays for Identification of Effector T-Cell Lineages
|
---|---|
Chapter number | 5 |
Book title |
Handbook of ELISPOT
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-8567-8_5 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8566-1, 978-1-4939-8567-8
|
Authors |
Jodi Hanson, Diana R. Roen, Paul V. Lehmann, Hanson, Jodi, Roen, Diana R., Lehmann, Paul V. |
Abstract |
Single color IFN-γ ELISPOT assays have evolved as a highly sensitive T cell immune monitoring platform. By detecting individual T cells that secrete IFN-γ in response to antigen exposure, these assays permit the measurement of the frequency of antigen-specific T cells among white blood cells. These assays therefore are well suited to assess clonal expansions, that is, whether a (Th1) T cell response has been induced to an antigen in a test subject. Single color IFN-γ ELISPOT assays are not suited, however, to provide information on the Th2/Th17 quality of the T cell response, nor do they provide insights into the differentiation state of CD8 cells. Recently it has been established that co-expression profiles of IL-2, TNF-α, and granzyme B along with IFN-γ permit to identify CD8 cell subpopulations. Naïve CD8 cells, central CD8 memory cells, CD8 terminal effector cells, polyfunctional CD8 cells, stem-cell like CD8 memory cells, dysfunctional- and senescent CD8 cells all differ in the extent they produce these molecules upon antigen re-encounter. We therefore have developed, and introduce here, a four color T cell ELISPOT assay in which the co-expression levels of IFN-γ, IL-2, TNF-α, and granzyme B can be established for individual antigen-specific CD8 cells, thereby identifying the activation/differentiation state of these cells. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 6 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Researcher | 2 | 33% |
Professor | 1 | 17% |
Unspecified | 1 | 17% |
Other | 1 | 17% |
Student > Master | 1 | 17% |
Other | 0 | 0% |
Readers by discipline | Count | As % |
---|---|---|
Immunology and Microbiology | 3 | 50% |
Agricultural and Biological Sciences | 2 | 33% |
Unspecified | 1 | 17% |