Measurement of Interleukins by ELISPOT Assay with Particular Application to Dual-Color Analysis (Stardust Assay)

The ordinary method for quantitative analysis of cytokines consists of measuring cytokines produced and accumulated in the supernatants of shortterm cultures, by means of enzyme immunoassay. However, this approach provides only cumulative amounts in a fixed time, thus limiting the sensitivity of the technique, particularly when relatively few cells are producing lymphokines of interest. In addition, this strategy does not permit an estimation of the frequency of corresponding secreting cells.