Octamer proteins inhibit IL-4 gene transcription in normal human CD4 T cells

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Abstract

The balance of Th1 (eg, interleukin-2 (IL-2)) and Th2 (eg, IL-4) cytokines produced by CD4 T cells markedly influences the outcome of the adaptive immune response. Although octamer transcription factor proteins increase IL-2 transcription in T cells, their role in IL-4 gene transcription remains controversial. We have previously shown and now confirm that the proximal octamer binding site of the human IL-4 promoter, which separates the two most proximal NFAT binding sites, is bound prior to, but not after, activation in vivo. Since these two NFAT sites are essential for optimal IL-4 promoter activity, this suggested that prior engagement by octamer proteins might prevent adjacent NFAT binding and inhibit IL-4 gene transcription. In support of this hypothesis, here we show that NFAT proteins are unable to bind to a combined octamer/NFAT site unless the octamer proteins are competed away. Moreover, activity of an IL-4 reporter gene mutated in the proximal octamer binding site is increased compared to the wild-type promoter in human peripheral blood CD4 T cells. In addition, over-expression of either Oct-1 or Oct-2 decreased wild-type IL-4 promoter activity, while increasing IL-2 promoter activity. No decrease in promoter activity was seen when Oct-1 or Oct-2 was over-expressed with the octamer-mutant IL-4 reporter gene. Thus, octamer proteins are candidates to promote a Th1 rather than Th2 pattern of cytokine gene expression by activated CD4 T cells.

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Acknowledgements

The authors would like to thank Dr Winship Herr (Cold Spring Harbor Laboratory, New York) and Dr Anjana Rao (Harvard University, Cambridge, MA) for the generous gifts of octamer expression plasmids and anti-NFAT1 antisera, 67.1, respectively. We also thank Dr Terri Finkel (Children’s Hospital of Philadelphia, PA) for critical reading of the manuscript, Dr Kate Sullivan (Children’s Hospital of Philadelphia) for helpful advice, and Yunxia Wang and Anna Genin for technical assistance.

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Correspondence to RQ Cron.

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Keywords

  • IL-4
  • octamer
  • transcription
  • human
  • T cell
  • interleukin-2

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