Macrophage-Specific NF-κB Activation Dynamics Can Segregate Inflammatory Bowel Disease Patients

Frontiers in Immunology

Vitor A. P. Martins Dos Santos¹  D. Mark Pritchard²  Chris S. Probert²  Barry J. Campbell²  Michael D. Burkitt²  Rachael Hough²  Werner Müller³  Lorraine Schmidt³  Stamatia Papoutsopoulou³  Michael H. R. White³  David G. Spiller³  Hazel England³  Pawel Paszek³  Dean A. Jackson³  François Bergey⁴  Gernot Sellge⁵ 

[1] Department of Agrotechnology and Food Sciences, Wageningen University, Wageningen, Netherlands;Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, United Kingdom;Faculty of Biology, Medicine and Health, School of Biological Sciences, University of Manchester, Manchester, United Kingdom;LifeGlimmer GmbH, Berlin, Germany;University Hospital RWTH Aachen, Aachen, Germany;

关键词: inflammatory bowel disease;    NF-κB;    macrophages;    cytokines;    Crohn's disease;    ulcerative colitis;   

DOI  :  10.3389/fimmu.2019.02168

来源: DOAJ

【 摘 要 】

The heterogeneous nature of inflammatory bowel disease (IBD) presents challenges, particularly when choosing therapy. Activation of the NF-κB transcription factor is a highly regulated, dynamic event in IBD pathogenesis. Using a lentivirus approach, NF-κB-regulated luciferase was expressed in patient macrophages, isolated from frozen peripheral blood mononuclear cell samples. Following activation, samples could be segregated into three clusters based on the NF-κB-regulated luciferase response. The ulcerative colitis (UC) samples appeared only in the hypo-responsive Cluster 1, and in Cluster 2. Conversely, Crohn's disease (CD) patients appeared in all Clusters with their percentage being higher in the hyper-responsive Cluster 3. A positive correlation was seen between NF-κB-induced luciferase activity and the concentrations of cytokines released into medium from stimulated macrophages, but not with serum or biopsy cytokine levels. Confocal imaging of lentivirally-expressed p65 activation revealed that a higher proportion of macrophages from CD patients responded to endotoxin lipid A compared to controls. In contrast, cells from UC patients exhibited a shorter duration of NF-κB p65 subunit nuclear localization compared to healthy controls, and CD donors. Analysis of macrophage cytokine responses and patient metadata revealed a strong correlation between CD patients who smoked and hyper-activation of p65. These in vitro dynamic assays of NF-κB activation in blood-derived macrophages have the potential to segregate IBD patients into groups with different phenotypes and may therefore help determine response to therapy.

【 授权许可】

Unknown