Cytokine Enzyme Linked Immunosorbent Spot (ELISPOT) Assay

This chapter provides an overview of cytokine enzyme linked immunosorbent spot (ELISPOT) assays. Versions of the ELISPOT assay have been used for nearly 20 years to detect antibody-secreting B cells and more recently cytokine-secreting lymphocytes. Several technical advances, including development of synthetic membranes and computer-assisted image analysis hardware and software, have improved the reliability and reproducibility of the assay and have facilitated data analysis. Cytokine ELISPOT analysis of T cell immunity has multiple advantages over many other approaches for accurately measuring the multiple relevant features of T cell immunity in animal models and in humans. This high-resolution assay has already proven to be invaluable in the research setting as an approach to delineating immune responses to infectious agents, tumor antigens, autoantigens, and transplant antigens. Similarly, ELISPOT assays are widely utilized in the investigation of a number of autoimmune diseases. Most of these studies are directed toward understanding the pathogenesis of disease and involve analysis of clonal size and cytokine-secretion profiles of autoantigen-specific T cells in the peripheral blood of patients with active autoimmune disease.