This assay is used to detect T cell response to specific antigenic stimulation. Stimulation can be performed with autologous or HLA-matched EBV-transformed B cells, PHA-blasted T cells, dendritic cells, or tumor cells. Cells are treated with an inhibitor of the secretory pathway permitting accumulation of cytokines inside the cytoplasm. Cytokine production is determined by staining the fixed and permeabilized cells. By combining ICS with staining for phenotypic markers and multimers, it is possible to determine the type of cells that produce the cytokine as well as the percentage of cytokine produced cell. This assay requires larger sample volumes than the ELISPOT. However, it permits the simultaneous determination of the phenotype of the cytokine-secreting cell as well as the identification of the type of cytokine being produced.
Please contact Dr. Junko Matsuzaki at (716) 845-8459 or Junko.Matsuzaki@roswellpark.org for a project specific cost estimate.