Dual-color ELISPOT assay for the simultaneous detection of IL-2 and/or IFN-γ secreting T cells

Salix Boulet, Michel L. Ndongala, Nicole F. Bernard

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The enzyme-linked immunospot (ELISPOT) assay measures the secretion intensity of effector molecules released by immune cells in response to ex vivo antigenic stimulation, as well as the frequency of these responding cells. This assay is highly sensitive, quantitative, easy to use, and amenable to high-throughput screening. For these reasons, the ELISPOT assay is considered by many as a gold standard for monitoring cellular immune responses. Until recently, ELISPOT assays using chromophores to detect the T cell secretion of cytokines were limited to the characterization of a single effector molecule. Notably, studies evaluating the immune response to chronic viral infections often measured IFN-γ secretion by ELISPOT because of the known antiviral effects of this cytokine as well as its correlation to the cytotoxic capacity of T cells. However, maintenance of both IFN-γ and IL-2 secretion by pathogen-specific T cells has been linked to a more favorable clinical outcome in human immunodeficiency virus (HIV) and Leishmania infections. Therefore, an ELISPOT assay able to simultaneously characterize T cell responses in terms of IL-2 and IFN-γ secretion is potentially relevant for the monitoring of immune responses to certain infectious agents. In this protocol, we describe an ELISPOT assay for the simultaneous detection of IL-2 and IFN-γ upon stimulation with viral peptides.

Original languageEnglish (US)
JournalCold Spring Harbor Protocols
Volume5
Issue number1
DOIs
StatePublished - 2010
Externally publishedYes

Fingerprint

Enzyme-Linked Immunospot Assay
T-cells
Interleukin-2
Assays
Color
T-Lymphocytes
Enzymes
Virus Diseases
Cytokines
Immunologic Monitoring
Leishmania
Molecules
Cellular Immunity
Monitoring
Antiviral Agents
Pathogens
Chromophores
Viruses
Maintenance
HIV

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Dual-color ELISPOT assay for the simultaneous detection of IL-2 and/or IFN-γ secreting T cells. / Boulet, Salix; Ndongala, Michel L.; Bernard, Nicole F.

In: Cold Spring Harbor Protocols, Vol. 5, No. 1, 2010.

Research output: Contribution to journalArticle

Boulet, Salix ; Ndongala, Michel L. ; Bernard, Nicole F. / Dual-color ELISPOT assay for the simultaneous detection of IL-2 and/or IFN-γ secreting T cells. In: Cold Spring Harbor Protocols. 2010 ; Vol. 5, No. 1.
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