Intracellular Cytokine Assays
This chapter provides an overview of intracellular cytokine assays. Intracellular cytokine assays are a relatively new method of identifying cytokine production by individual T cells and have the ability to correlate cytokine expression with cell surface phenotype without cell separation. In addition, this highly sensitive flow cytometric method allows for the rapid detection of low frequency T cells expressing cytokine in response to specific antigen stimulation. The unique capabilities of this method make it a model assay for clinical and research applications. The overall premise of intracellular cytokine assays is direct detection of intracellular cytokine expression in response to antigen stimulation. Intracellular cytokine assays can be performed using various sources of cells and antigen depending on the target(s) of interest. T cell stimulation can be performed directly on whole blood, peripheral blood mononuclear cells, in vitro manipulated lymphocytes, isolated cells, and lymph nodes; although using whole blood for these assays provides a more physiological environment and may have an effect on the T cell response to stimulation. Since these assays are most often used to detect very low frequency events, appropriate control antigens are particularly important to ensure clear antigen specific response. © 2005 Elsevier Ltd. All rights reserved.
