Impact of human sepsis on CCCTC-binding factor associated monocyte transcriptional response of Major Histocompatibility Complex II components

PloS One
Benedikt Hermann SieglerSebastian Weiterer

Abstract

Antigen presentation on monocyte surface to T-cells by Major Histocompatibility Complex, Class II (MHC-II) molecules is fundamental for pathogen recognition and efficient host response. Accordingly, loss of Major Histocompatibility Complex, Class II, DR (HLA-DR) surface expression indicates impaired monocyte functionality in patients suffering from sepsis-induced immunosuppression. Besides the impact of Class II Major Histocompatibility Complex Transactivator (CIITA) on MHC-II gene expression, X box-like (XL) sequences have been proposed as further regulatory elements. These elements are bound by the DNA-binding protein CCCTC-Binding Factor (CTCF), a superordinate modulator of gene transcription. Here, we hypothesized a differential interaction of CTCF with the MHC-II locus contributing to an altered monocyte response in immunocompromised septic patients. We collected blood from six patients diagnosed with sepsis and six healthy controls. Flow cytometric analysis was used to identify sepsis-induced immune suppression, while inflammatory cytokine levels in blood were determined via ELISA. Isolation of CD14++ CD16-monocytes was followed by (i) RNA extraction for gene expression analysis and (ii) chromatin immunoprecipitation to a...Continue Reading

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Citations

May 13, 2020·Epigenomics·Jesús Beltrán-GarcíaJosé Luis García-Giménez
Jul 30, 2021·Frontiers in Medicine·Renata Brito Falcão-HolandaReinaldo Salomão

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Methods Mentioned

BETA
acetylation
Flow cytometry
density gradient centrifugation
PCR
immunoprecipitation
enzyme-linked immunosorbent assay
ELISA
Assay
ChIP-seq
histone acetylation

Software Mentioned

GraphPad Prism
GraphPad

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