T Cell ELISPOT: For the Identification of Specific Cytokine-Secreting T Cells

Kerry M Leehan1, Kristi A Koelsch

  • 1Arthritis and Clinical Immunology Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK, USA.

The ELISPOT is a powerful functional assay used to detect biological activity and immunological secretions from immune cells. In this chapter, we specifically discuss T cell ELISPOT methods for the detection of secreted cytokines. A detailed protocol is given enabling the detection of interferon gamma-secreting CD8(+) T cells and/or peripheral blood mononuclear cells following their isolation and polyclonal activation. Included is a brief discussion on choosing the activation method for your T cell ELISPOT assay, as well as additional instructions for the adaptation of this protocol for the study of memory and antigen-specific T cell responses.