Characterization of a novel PMA-inducible pathway of interleukin-13 gene expression in T cells

Immunology. 2006 Jan;117(1):29-37. doi: 10.1111/j.1365-2567.2005.02260.x.

Abstract

Although interleukin 13 (IL-13) is an important mediator of asthma and allergic diseases, the molecular mechanisms regulating IL-13 gene expression are not well understood. This study was designed to define the molecular mechanisms governing IL-13 gene expression in T cells. IL-13 expression was examined in human peripheral blood T cells and in the EL-4 T-cell line by enzyme-linked immunosorbent assay and reverse-transcription polymerase chain reaction. An IL-13 promoter deletion analysis was performed using luciferase-based reporter plasmids transiently transfected into EL-4 cells by electroporation. DNA binding factors were investigated using electrophoretic mobility shift assays. In contrast to IL-4 expression, which required concomitant activation of calcium- and protein kinase C- (PKC-) dependent signalling pathways, PKC activation alone was sufficient for IL-13 protein secretion in mitogen-primed (but not resting) peripheral blood T cells, and for IL-13 mRNA expression and promoter activity in EL-4 T cells. Promoter deletion analysis localized a phorbol 12-myristate 13-acetate (PMA)-sensitive element to a proximal promoter region between -109 and -79 base pairs upstream from the IL-13 transcription start site. This promoter region supported the binding of both constitutive and PMA-inducible nuclear factors in gel shift assays.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • Cells, Cultured
  • Gene Expression Regulation / immunology
  • Humans
  • Interleukin-13 / genetics
  • Interleukin-13 / immunology
  • Interleukin-13 / metabolism*
  • Mice
  • Molecular Sequence Data
  • Promoter Regions, Genetic
  • Protein Kinase C / immunology
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Signal Transduction / immunology
  • Species Specificity
  • T-Lymphocytes / immunology*
  • Tetradecanoylphorbol Acetate / immunology*
  • Transcription, Genetic / immunology

Substances

  • Interleukin-13
  • Protein Kinase C
  • Tetradecanoylphorbol Acetate