Retroviral transduction and reporter assay: Transcription factor cooperation in Th9 cell development

Abstract

Naïve CD4+ T cells differentiate into different T helper subsets in response to specific cytokine environment and transcription factors. Th9 cells are induced in response to signals from cytokines, TGF-β and IL-4. Transcription factors that are downstream of these cytokines converge to drive the development of Th9 cells. Retroviral transduction allows the genetic modification in T cells thereby helping us to better understand the molecular mechanisms that control their development as well as function. In this chapter, an optimized protocol for retroviral transduction of murine Th9 cells as well as transient transfection of Th9 cells with luciferase reporter constructs is described.