IL-9 is a pro-inflammatory cytokine implicated in certain immune-mediated diseases where chronic or acute inflammation of the mucosa plays an important role. Although initially described as being produced by what was then thought to be Th2 cells, it was later described that specialized lymphocyte populations are involved in IL-9 production. In addition to the classical Th9 effector (subset of CD4+ T cells), IL-9 is also produced by nonconventional lymphocytes, namely invariant natural killer T (iNKT) cells and innate lymphoid cells (ILCs). The identification of IL-9-producing cells by flow cytometry and cytokine measurements are pivotal for assigning and defining functional cellular phenotypes. In this chapter we provide methods for the in vitro polarization of IL-9-producing nonconventional lymphocytes and the best conditions for the detection of IL-9 production by intracellular staining.
Almeida, S. C. P., & Graca, L. (2017). IL-9 production by nonconventional T helper cells. In Methods in Molecular Biology (Vol. 1585, pp. 93–109). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6877-0_8