Rapid protocol for in vitro induction of cytokine associated with Th2 and Th17 cells from PBMC

Abstract

Abstract T helper cell is a key component of the immune system. They mediate humoral and cellular immune responses against pathogens and other diseases like cancer by inducing and activating effector cells such as B cells, cytotoxic T cells and innate cells. They are also involved in the regulation of immune responses by controlling the persistence and magnitude of the response. Activation of naïve T cell is initiated by interaction with antigen presenting cells. The downstream signal transduction and the cytokine milieu can lead to their proliferation and differentiation into different T cell subtypes. To facilitate the study of Th2 and Th17 cells, we present a rapid protocol to efficiently induce cytokine associated with Th2 and Th17 cells from total PBMC. Canonical anti-CD3 and anti-CD28 clones are used to activate the purified cells. Different sets of bioassay-validated cytokines, including IL-12, IL-4, IL-6, IL-1β, IL-2, and TGF-β1 are used to induce cytokine associated with Th2 and Th17 cells. Hallmark cytokine and transcription factor expression profile of different lineages are evaluated by Flow Cytometry. In summary, we have successfully developed a rapid protocols to induce cytokine expression in vitro associated with Th2 and Th17 using high quality reagents developed in BioLegend.