Home » Key Scientific Articles » Multiple interferon regulatory factor and NF-κB sites cooperate in mediating cell-type- and maturation-specific activation of the human CD83 promoter in dendritic cells.

Multiple interferon regulatory factor and NF-κB sites cooperate in mediating cell-type- and maturation-specific activation of the human CD83 promoter in dendritic cells.

Stein MF, Lang S, Winkler TH, Deinzer A, Erber S, Nettelbeck DM, Naschberger E, Jochmann R, Stürzl M, Slany RK, Werner T, Steinkasserer A, Knippertz I.

Mol Cell Biol. 2013 Apr;33(7):1331-44.

Department of Dermatology, University Hospital Erlangen, Erlangen, Germany.

 

Abstract

 

CD83 is one of the best-known surface markers for fully mature dendritic cells (mature DCs), and its cell-type- and maturation-specific regulation makes the CD83 promoter an interesting tool for the genetic modulation of DCs. To determine the mechanisms regulating this DC- and maturation-specific CD83 expression, chromatin immunoprecipitation (ChIP)-on-chip microarray, biocomputational, reporter, electrophoretic mobility shift assay (EMSA), and ChIP analyses were performed. These studies led to the identification of a ternary transcriptional activation complex composed of an upstream regulatory element, a minimal promoter, and an enhancer, which have not been reported in this arrangement for any other gene so far. Notably, these DNA regions contain a complex framework of interferon regulatory factor (IRF)- and NF-κB transcription factor-binding sites mediating their arrangement. Mutation of any of the IRF-binding sites resulted in a significant loss of promoter activity, whereas overexpression of NF-κB transcription factors clearly enhanced transcription. We identified IRF-1, IRF-2, IRF-5, p50, p65, and cRel to be involved in regulating maturation-specific CD83 expression in DCs. Therefore, the characterization of this promoter complex not only contributes to the knowledge of DC-specific gene regulation but also suggests the involvement of a transcriptional module with binding sites separated into distinct regions in transcriptional activation as well as cell-type- and maturation-specific transcriptional targeting of DCs.

 

 

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Additional information

One of the biggest challenges facing cancer therapy today is the generation of cell-specific treatment strategies. Towards this goal, dendritic cells (DCs) represent a promising tool for cancer immunotherapy, especially since DCs are the only antigen-presenting cells (APCs) known today, able to induce naïve T cells responses. Moreover, ex vivo generated DCs have proven effective in stimulation of tumor specific immune responses not only in vitro, but also in vivo in vaccinated cancer patients. However, there is still a high medical need to develop new treatment modalities and DC-based cancer vaccines, e.g. by transcriptional targeting of DCs directly within cancer patients, without the need of a work-, time- and cost intensive ex vivo generation of DC-vaccines as currently used in most clinical DC-trials. In order to selectively express tumor-specific antigens only in immunogenic mature DCs, and not in tolerogenic immature DCs, of tumor patients, this newly characterized human CD83 promoter represents an ideal tool for transcriptional in vivo targeting directly within patients. Because of its specificity for mature DCs it will set the stage for next generation in vivo vaccination strategies that should be particularly effective and safe as it for the first time assures selective antigen expression in mature DCs while avoiding expression in tolerogenic immature DCs. (see also European Patent application: EP11164344.1; PCT patent application: PCT/EP2012/057760)

 

Figure legend:

Next generation DC-vaccination strategy directly in tumor patients. Tumor antigens will be expressed under the control of the CD83 promotor in mature, immunogenic DC, directly in tumor patients. The long term aim of this new in vivo DC-targeting strategy is the induction of protective anti-tumoral immune responses directly in cancer patients, without the need of the work-, time- and cost intensive ex vivo generation of DC-vaccines, as currently used in state of the art clinical DC-trials.

 

Antw Multiple Interferon Regulatory Factor and