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

Stamatia Papoutsopoulou*, Michael D. Burkitt, François Bergey, Hazel England, Rachael Hough, Lorraine Schmidt, David G. Spiller, Michael H.R. White, Pawel Paszek, Dean A. Jackson, Vitor A.P. Martins Dos Santos, Gernot Sellge, D.M. Pritchard, Barry J. Campbell, Werner Müller, Chris S. Probert

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

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.
Original languageEnglish
Article number2168
Number of pages11
JournalFrontiers in Immunology
Volume10
DOIs
Publication statusPublished - 11 Sep 2019

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Inflammatory Bowel Diseases
Macrophages
Crohn Disease
Luciferases
Cytokines
Ulcerative Colitis
Lipid A
Lentivirus
Endotoxins
Blood Cells
Transcription Factors
Tissue Donors
Phenotype
Biopsy
Therapeutics
Serum

Cite this

Papoutsopoulou, S., Burkitt, M. D., Bergey, F., England, H., Hough, R., Schmidt, L., ... Probert, C. S. (2019). Macrophage-Specific NF-κB Activation Dynamics Can Segregate Inflammatory Bowel Disease Patients. Frontiers in Immunology, 10, [2168]. https://doi.org/10.3389/fimmu.2019.02168
Papoutsopoulou, Stamatia ; Burkitt, Michael D. ; Bergey, François ; England, Hazel ; Hough, Rachael ; Schmidt, Lorraine ; Spiller, David G. ; White, Michael H.R. ; Paszek, Pawel ; Jackson, Dean A. ; Martins Dos Santos, Vitor A.P. ; Sellge, Gernot ; Pritchard, D.M. ; Campbell, Barry J. ; Müller, Werner ; Probert, Chris S. / Macrophage-Specific NF-κB Activation Dynamics Can Segregate Inflammatory Bowel Disease Patients. In: Frontiers in Immunology. 2019 ; Vol. 10.
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abstract = "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.",
author = "Stamatia Papoutsopoulou and Burkitt, {Michael D.} and Fran{\cc}ois Bergey and Hazel England and Rachael Hough and Lorraine Schmidt and Spiller, {David G.} and White, {Michael H.R.} and Pawel Paszek and Jackson, {Dean A.} and {Martins Dos Santos}, {Vitor A.P.} and Gernot Sellge and D.M. Pritchard and Campbell, {Barry J.} and Werner M{\"u}ller and Probert, {Chris S.}",
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Papoutsopoulou, S, Burkitt, MD, Bergey, F, England, H, Hough, R, Schmidt, L, Spiller, DG, White, MHR, Paszek, P, Jackson, DA, Martins Dos Santos, VAP, Sellge, G, Pritchard, DM, Campbell, BJ, Müller, W & Probert, CS 2019, 'Macrophage-Specific NF-κB Activation Dynamics Can Segregate Inflammatory Bowel Disease Patients', Frontiers in Immunology, vol. 10, 2168. https://doi.org/10.3389/fimmu.2019.02168

Macrophage-Specific NF-κB Activation Dynamics Can Segregate Inflammatory Bowel Disease Patients. / Papoutsopoulou, Stamatia; Burkitt, Michael D.; Bergey, François; England, Hazel; Hough, Rachael; Schmidt, Lorraine; Spiller, David G.; White, Michael H.R.; Paszek, Pawel; Jackson, Dean A.; Martins Dos Santos, Vitor A.P.; Sellge, Gernot; Pritchard, D.M.; Campbell, Barry J.; Müller, Werner; Probert, Chris S.

In: Frontiers in Immunology, Vol. 10, 2168, 11.09.2019.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Macrophage-Specific NF-κB Activation Dynamics Can Segregate Inflammatory Bowel Disease Patients

AU - Papoutsopoulou, Stamatia

AU - Burkitt, Michael D.

AU - Bergey, François

AU - England, Hazel

AU - Hough, Rachael

AU - Schmidt, Lorraine

AU - Spiller, David G.

AU - White, Michael H.R.

AU - Paszek, Pawel

AU - Jackson, Dean A.

AU - Martins Dos Santos, Vitor A.P.

AU - Sellge, Gernot

AU - Pritchard, D.M.

AU - Campbell, Barry J.

AU - Müller, Werner

AU - Probert, Chris S.

PY - 2019/9/11

Y1 - 2019/9/11

N2 - 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.

AB - 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.

U2 - 10.3389/fimmu.2019.02168

DO - 10.3389/fimmu.2019.02168

M3 - Article

VL - 10

JO - Frontiers in Immunology

JF - Frontiers in Immunology

SN - 1664-3224

M1 - 2168

ER -