In Vivo Anergized CD4+ T Cells Have Defective Expression and Function of the Activating Protein-1 Transcription Factor

Abstract

The transcription factor activating protein-1 (AP-1) contributes significantly to the regulation of IL-2 gene expression during T cell activation and has been suggested to play a unique role in T cell anergy in vitro. In this study we have used the superantigen staphylococcal enterotoxin A to investigate the regulation of AP-1 in T cell anergy in vivo. Repeated injections of staphylococcal enterotoxin A induce a state of anergy in CD4+ T cells, characterized by reduced expression of IL-2 at mRNA and protein levels. The perturbed IL-2 response in anergic T cells correlated with reduced DNA binding activity of the transcription factors AP-1 and Fos/Jun-containing NF-AT. Using AP-1-luciferase reporter transgenic mice, we now demonstrate the lack of AP-1-dependent transcription. AP-1 activity is controlled by synthesis of its subunits Fos and Jun and by posttranslational phosphorylations. Analysis of Fos and Jun protein levels revealed no major differences in the expression of Jun proteins, but a marked decrease in c-Fos in anergic T cells. Experiments in transgenic mice overexpressing c-Fos (H2-c-fos) showed reconstituted AP-1 DNA binding. In contrast, the AP-1-driven transcription and IL-2 production remained suppressed. The Jun N-terminal kinase is known to play a critical role in regulating AP-1 trans-activation. Analyses of Jun N-terminal kinase demonstrated normal protein amounts, but reduced enzymatic activity, in anergic compared with activated CD4+ T cells. This suggests that in vivo anergized T cells have defects in the AP-1 pathway due to both reduced protein expression and perturbed posttranslational modifications.