title: Epigenetic Regulation in Early Childhood: A Miniaturized and Validated Method to Assess Histone Acetylation.
abstract: BACKGROUND
  Chronic inflammatory diseases including allergies and asthma are the result of complex interactions between genes and environmental factors. Epigenetic mechanisms comprise a set of biochemical reactions that regulate gene expression. In order to understand the cause-effect relationship between environmental exposures and disease development, methods capable of assessing epigenetic regulation (also) in large cohorts are needed.
  
  
  METHODS
  For this purpose, we developed and evaluated a miniaturized chromatin immunoprecipitation (ChIP) assay allowing for a cost-effective assessment of histone acetylation of candidate genes in a quantitative fashion. This method was then applied to assess H3 and H4 histone acetylation changes in cord blood (CB) samples from an established cohort of Australian children exposed in the fetal period to either very low or very high levels of maternal folate.
  
  
  RESULTS
  Our ChIP assay was validated for a minimum requirement of 1 × 105 target cells (e.g. CD4+ T cells). Very high levels of maternal folate were significantly associated with increased H3/H4 acetylation at GATA3 and/or IL9 promoter regions in CD4+ T cells in CB.
  
  
  CONCLUSIONS
  We developed a ChIP method allowing reliable assessment of H3/H4 acetylation using 1 × 105 cells only. Practical application of this assay demonstrated an association between high maternal folate exposure and increased histone acetylation, corresponding to a more transcriptionally permissive chromatin status in the promoter regions of some Th2-related genes.
authors.affiliation: Institute of Laboratory Medicine, Pathobiochemistry and Molecular Diagnostics, Philipps University Marburg, Marburg, Germany.
authors.forename: Hani
authors.initials: H
authors.lastname: Harb
authors.forename: Manori
authors.initials: M
authors.lastname: Amarasekera
authors.forename: Sarah
authors.initials: S
authors.lastname: Ashley
authors.forename: Meri K
authors.initials: MK
authors.lastname: Tulic
authors.forename: Petra Ina
authors.initials: PI
authors.lastname: Pfefferle
authors.forename: Daniel P
authors.initials: DP
authors.lastname: Potaczek
authors.forename: David
authors.initials: D
authors.lastname: Martino
authors.forename: Dörthe A
authors.initials: DA
authors.lastname: Kesper
authors.forename: Susan L
authors.initials: SL
authors.lastname: Prescott
authors.forename: Harald
authors.initials: H
authors.lastname: Renz
mesh_terms: D000107:Acetylation; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D002648:Child; D047369:Chromatin Immunoprecipitation; D044127:Epigenesis, Genetic; D005492:Folic Acid / Q000097:blood; D050990:GATA3 Transcription Factor / Q000235:genetics; D006657:Histones / Q000378:metabolism*; D006801:Humans; D057230:Limit of Detection; D012016:Reference Values
chemical_list: D050990:GATA3 Transcription Factor; C494749:GATA3 protein, human; D006657:Histones; D005492:Folic Acid