The Ex Vivo IFN-γ Enzyme-Linked Immunospot (ELISpot) Assay
The quantification of single cell interferon-gamma (IFN-γ) release for assessing cellular immune responses using the Enzyme-linked immunospot (ELISPOT) assay is an invaluable technique in immunology. Peripheral blood mononuclear cells (PBMC) are stimulated in vitro with recombinant proteins, peptides and recently whole malaria organisms. Stimulation may be short term (20–36 h) or long term (cultured ELISpot, up to 7 days). ELISpot is also able to quantify other cytokines secreted by antigen-specific T-cells, such as interleukin-2, interleukin-5, and other interleukins. ELISpot is playing an important role especially in vaccine research studies.
Key wordsELISPOT Peripheral blood mononuclear cells PBMC Interferon-gamma IFN-γ Cytokines T cells
The following have made major contributions to the development and use of this assay: Harini Ganeshan, Maria Belmonte, Jun Huang, Esteban Abot, and Arnel Belmonte, and helped write this chapter with Michael R. Hollingdale.
- 5.Cox JH et al (2002) Accomplishing cellular immune assays for evaluation of vaccine efficacy in developing countries. In: Rose NR, Hamilton RG, Detrick B (eds) Manual clinical laboratory immunology. ASM Press, Washington, DC, pp 301–315Google Scholar