Primary Human CD4+ T Cells Contain Heterogeneous IκB Kinase Complexes: Role in Activation of the IL-2 Promoter

Abstract

AbstractNF-κB transcription factors play an important role in the activation of the IL-2 gene in response to TCR ligation. The release of NF-κB factors to the nucleus requires phosphorylation and degradation of the inhibitory κ-B proteins (IκBs). IκBα and IκBβ phosphorylation is dependent on dual signaling by the TCR and the CD28 accessory receptor. This pathway involves a multisubunit IκB kinase (IKK) complex, which includes the IKKα (IKK-1) and IKKβ (IKK-2) kinases. We demonstrate that stimulation of primary human CD4+ T cells by CD3/CD28 activates two distinct endogenous IKK complexes, a heterodimeric IKKα/β and a homodimeric IKKβ complex. IKKβ overexpression in a Jurkat cell line resulted in the formation of a constitutively active IKK complex, which was CD3/CD28 inducible. In contrast, ectopic expression of IKKα assembled into a complex with negligible IκB kinase activity. Moreover, IKKβ, but not IKKα, overexpression enhanced transcriptional activation of the CD28 response element in the IL-2 promoter. Conversely, only kinase-inactive IKKβ interfered in the activation of the IL-2 promoter. Sodium salicylate, an inhibitor of IKKβ, but not IKKα, activity, inhibited IL-2 promoter activation as well as IL-2 secretion and interfered in activation of both the heterodimeric as well as the homodimeric IKK complexes in primary CD4+ T cells. Taken together, these data demonstrate the presence of an IKKβ-mediated signaling pathway that is activated by TCR and CD28 coligation and regulates IL-2 promoter activity.