Functional Analysis of Histone Deacetylase 11 (HDAC11)

Methods Mol Biol. 2016:1436:147-65. doi: 10.1007/978-1-4939-3667-0_11.

Abstract

The physiological role of histone deacetylase 11 (HDAC11), the newest member of the HDAC family, remained largely unknown until the discovery of its regulatory function in immune cells. Among them, the regulation of cytokine production by antigen-presenting cells and the modulation of the suppressive ability of myeloid-derived suppressor cells (MDSCs) (Sahakian et al. Mol Immunol 63: 579-585, 2015; Wang et al. J Immunol 186: 3986-3996, 2011; Villagra et al. Nat Immunol 10: 92-100, 2009). Our earlier data has demonstrated that HDAC11, by interacting at the chromatin level with the IL-10 promoter, downregulates il-10 transcription in both murine and human APCs in vitro and ex vivo models (Villagra et al. Nat Immunol 10: 92-100, 2009). However the role of HDAC11 in other cell types still remains unknown. Here we present several methods that can potentially be used to identify the functional role of HDAC11, assigning special attention to the evaluation of immunological parameters.

Keywords: Chromatin immunoprecipitation; Flow cytometry; Genotyping; Histone deacetylase 11; PCR; Phenotyping; qRT-PCR.

MeSH terms

  • Animals
  • Antigen-Presenting Cells / cytology
  • Antigen-Presenting Cells / immunology
  • Chromatin Immunoprecipitation / methods*
  • Cytokines / genetics*
  • Cytokines / metabolism
  • Flow Cytometry
  • Gene Expression Regulation
  • Histone Deacetylases / genetics
  • Histone Deacetylases / metabolism*
  • Mice
  • Myeloid-Derived Suppressor Cells / cytology
  • Myeloid-Derived Suppressor Cells / immunology
  • Promoter Regions, Genetic

Substances

  • Cytokines
  • Hdac11 protein, mouse
  • Histone Deacetylases