Flow Cytometric Assessment of STAT Molecules in Th9 Cells.

IL-9-producing Th9 cells are a novel subset of T helper cells that develop independently of other T helper subsets. Th9 cells have been implicated in the pathogenesis of allergic asthma and autoimmunity, while also serving as critical effector T cells in mediating antitumor immune responses. Concomitant presence of TGF-ß and IL-4 lead to the differentiation of naïve CD4+ T cells towards the Th9 phenotype. In addition, several cytokines, including IL-1ß, IL-2, IL-25, and IL-33, further amplify Th9 responses. Negative regulators of Th9 cells include other cytokines such as IFN-γ, IL-23, and IL-27. Here, we describe a detailed protocol for the analysis of STAT molecules involved in the differentiation of Th9 cells and Th9 inhibition by IL-27.