Hovhannisyan et al. first showed evidence of plasticity between Treg and Th17 in the inflamed intestine of Crohn’s disease (CD) patients. Our previous report suggests that the inflammatory cytokine milieu generates IL-17+ Foxp3+ CD4+ T lymphocytes which is a crossover population converting Treg subset to Th17 in the peripheral blood of IBD patients. This is considered as an evidence of Treg/Th17 plasticity.
The aim of this study was to characterize a variety of helper T cell crossover population, not limited to IL-17+ Foxp3+ CD4+ T lymphocytes, in the lamina propria (LP) of IBD patients.
Fresh colonoscopic biopsies were obtained from patients with CD (n = 50) and ulcerative colitis (UC, n = 32) and from healthy controls (HC, n = 25). LP mononuclear cells were assessed for intracellular cytokines and transcription factors such as IFNγ, IL-13, IL-17, IL-22, T-bet, Gata-3, RORγt, and Foxp3 using multicolor flow cytometry to detect subsets of LP CD4+ T lymphocytes.
Patients with IBD demonstrated increased crossover populations in IL-17+ Foxp3+, T-bet+ Foxp3+, Gata3+ Foxp3+, RORγt+ Foxp3+ populations compared to HC. There was an inverse correlation of Harvey–Bradshaw index with Gata3+ Foxp3+ population in CD patients, while IL-13+ Foxp3+ population was directly correlated with Mayo clinical scores in UC patients. Furthermore, total IL-22 expressing cells as well as Th22 and IL-22+ Th1 populations were decreased in UC compared to CD and HC.
IBD patients exhibit the increased crossover populations in LP Treg cells toward Th2 and Th17 compared to HC. The prevalence of Treg/Th2 crossover populations is associated with clinical disease score of IBD.
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Inflammatory bowel diseases
T helper type 1 cells
T helper type 2 cells
T helper type 17 cells
T helper type 22 cells
Tumor necrosis factor
Regulatory T helper cells
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The authors thank to the Alberta IBD consortium for financial support; Ms. Gurmeet Bindra in the inflammatory intestinal tissue bank (IITB) and the staff in IBD clinic for assistance with research sample collection; Dr. Karen Poon (Nicole Perkins Microbe Core Facility, Calvin, Phoebe and Joan Snyder Institute for Chronic Disease) and Mr. Ronald Chan for assistance with data acquisition; Drs. Tie Wang, Donald Buie, Anthony MacLean, and Iwona Auer-Grzesiak for assistance with optimizing study design; and all patients and healthy individuals who participated into this study.
Conflict of interest
The authors declare that they have no conflict of interest.
Ji Li and Aito Ueno share the first authorship.
Xianyong Gui and Subrata Ghosh share the senior authorship.
Study Sponsor Alberta IBD Consortium/Alberta Innovates—Health Solutions.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Flow cytometry plots. The gating strategy is shown in (A). First, the acquired data were gated lymphocytes in FSC-A vs. SSC-A display, followed by doublets discrimination in FSC-A vs. FSC-H display, and then live/dead discrimination. Live CD4+ T cells are further analyzed. Representative data of CD, UC, and HC are shown in B-D. The upper panels display Foxp3 (X-axis) vs. IFNγ (Y-axis) (B), the middle panels Foxp3 (X-axis) vs. IL-17 (Y-axis) display (C) and the lower panels IL-17 (X-axis) versus IFNγ (Y-axis) (D). (PPTX 113 kb)
Prevalence of Th subsets in lamina propria mononuclear cells. The prevalence of expressing a Th1 signature cytokine, IFNγ (A), a Th1 transcription factor, T-bet (B), a Th2 signature cytokine, IL-13 (C), a Th2 transcription factor, Gata3 (D), a Th17 signature cytokine, IL-17A (E), a Th17 transcription factor RORγt (F), a Treg transcription factor Foxp3 (G), and a combination of CD25 and Foxp3 (H) in CD4 + T cells are shown in this figure. * p < 0.05, **p < 0.01, *** p < 0.001, **** P < 0.0001. (PPTX 99 kb)
Concordance between endoscopic scores (SES-CD scores) and Foxp3+ crossover populations in lamina propria mononuclear cells of CD patients. The prevalence of IFNγ+ Foxp3+ (A), IL-13+ Foxp3+ (B), IL-17+ Foxp3+ (C), T-bet+ Foxp3+ (D), Gata3+ Foxp3+ (E), RORγt+ Foxp3+ (F), and IFNγ+ IL-17+ (G) is shown in this figure. (PPTX 146 kb)
Concordance between Mayo endoscopic subscores and CD4+ T cell crossover populations in lamina propria mononuclear cells of UC patients. The prevalence of IFNγ+ Foxp3+ (A), IL-13+ Foxp3+ (B), IL-17+ Foxp3+ (C), T-bet+ Foxp3+ (D), Gata3+ Foxp3+ (E), RORγt+ Foxp3+ (F), and IFNγ+ IL-17+ (G) is shown in this figure. * p < 0.05 (PPTX 102 kb)
Association between IL-22+ T helper cell subsets and clinical disease activity scores evaluated by Harvey–Bradshaw index in CD (A) and Mayo clinical index scores in UC cohort (B). Total IL-22+, IL-22+ IL-17+ (IL-22+ Th17), IL-22+ IFN+ (IL-22+ Th1) and IL-22+ IL-17-IFNγ-(Th22) cells are shown in this figure. (PPTX 81 kb)
Association between IL-22+ T helper cell subsets and endoscopic disease activity evaluated by SES-CD scores in CD cohort (A) and Mayo endoscopic subscores in UC cohort (B). Total IL-22+, IL-22+ IL-17+ (IL-22+ Th17), IL-22+ IFN+ (IL-22+ Th1) and IL-22+ IL-17-IFNγ-(Th22) cells are shown in this figure. (PPTX 80 kb)
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Li, J., Ueno, A., Iacucci, M. et al. Crossover Subsets of CD4+ T Lymphocytes in the Intestinal Lamina Propria of Patients with Crohn’s Disease and Ulcerative Colitis. Dig Dis Sci 62, 2357–2368 (2017). https://doi.org/10.1007/s10620-017-4596-9
- T cell crossover population
- Crohn’s disease
- Ulcerative colitis
- Regulatory T cells