KLRG1 and CD103 Expressions Define Distinct Intestinal Tissue-Resident Memory CD8 T Cell Subsets Modulated in Crohn's Disease - PubMed (original) (raw)

KLRG1 and CD103 Expressions Define Distinct Intestinal Tissue-Resident Memory CD8 T Cell Subsets Modulated in Crohn's Disease

Hugo Bottois et al. Front Immunol. 2020.

Abstract

Intestinal tissue-resident memory CD8 T cells (Trm) are non-recirculating effector cells ideally positioned to detect and react to microbial infections in the gut mucosa. There is an emerging understanding of Trm cell differentiation and functions, but their implication in inflammatory bowel diseases, such as Crohn's disease (CD), is still unknown. Here, we describe CD8 cells in the human intestine expressing KLRG1 or CD103, two receptors of E-cadherin. While CD103 CD8 T cells are present in high numbers in the mucosa of CD patients and controls, KLRG1 CD8 T cells are increased in inflammatory conditions. Mucosal CD103 CD8 T cells are more responsive to TCR restimulation, but KLRG1 CD8 T cells show increased cytotoxic and proliferative potential. CD103 CD8 T cells originate mostly from KLRG1 negative cells after TCR triggering and TGFβ stimulation. Interestingly, mucosal CD103 CD8 T cells from CD patients display major changes in their transcriptomic landscape compared to controls. They express Th17 related genes including CCL20, IL22, and IL26, which could contribute to the pathogenesis of CD. Overall, these findings suggest that CD103 CD8 T cells in CD induce a tissue-wide alert increasing innate immune responses and recruitment of effector cells such as KLRG1 CD8 T cells.

Keywords: CD103; Crohn's disease; IBD – inflammatory bowel diseases; KLRG1; T cells; TRM cells.

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Figures

Figure 1

CD103 and KLRG1 expressions on CD8 T cells are mutually exclusive in blood and intestinal mucosa. (A) Representative staining and (B) proportions of CD4 and CD8 T cell subsets in blood and ileal mucosa of control individual (CT) (n = 25 and n = 10 for blood and ileum, respectively; boxes represent median and range). (C) Representative staining and (D) quantification of CD103, KLRG1 and NKG2A expression on CD8 T cells in blood and ileal mucosa of control individuals (CT) (n = 25 and n = 10 for blood and ileum, respectively; boxes represent median and range; unpaired _t_-test: **p < 0.001, ****p < 0.00001). (E) tSNE analysis of CD103, KLRG1 and NKG2A expression on ileal mucosal CD8 T cells.

Figure 2

TGFβ and TCR triggering highly induces CD103 expression on T cells, but decrease KLRG1 expression. (A) Representative expression of CD103 and KLRG1 on CD8 T cells following IL-15 and TGFβ incubation and TCR stimulation during 7 days. (B) Quantification and (C) proportion of CD103 and KLRG1 expression on CD8 T cells (dots and bars represent 5 independent experiments with mean; ANOVA with Bonferoni post-test:*p < 0.05, **p < 0.001).

Figure 3

KLRG1-CD8 T cells acquire CD103 following TGFβ exposure and TCR triggering. (A) Representative expression of C0103 and NKG2A on sorted memory KLRG1- and KLRG1+ after incubation with or without TGFβ and TCR triggering. (B) Quantification of C0103, KLRG1, and C0103+NKG2A+ expression on C08 T cells (dots and bars represent 4 independent experiments with mean and SEM; ANOVA with Bonferoni post-test: *p < 0.05, ***p < 0.0001).

Figure 4

CD103 and KLRG1 CD8 Trm cell subsets in ileal Crohn's disease. (A) Representative CD4 and CD8 T cells staining and (B) quantification in the ileal mucosa of control individuals (CT; n = 10), CD patients in macroscopically inflamed (lnf) and non-inflamed (non-inf) segments of the ileocecal resections (n = 69; boxes represent median and range; unpaired _t_-test: not significant). (C) Representative expression and (D) quantification of CD103 (n = 69), KLRG1 (n = 25), and NKG2A (n = 35) on CD8 T cells from ileal mucosa as in A (bars represent means; unpaired _t_-test between CT and CD; paired _t_-test between lnf and Non-lnf: *p < 0.05, **p < 0.001).

Figure 5

Mucosal CD103+ CD8 T cells from CD patient are more responsive to TCR triggering than CD103- CD8 T cells. (A) Representative expression of CD103 and CD25 activation marker on mucosal CD8 T cells from CD patients after TCR triggering. (B) Quantification of CD25 expression on CD103+ and CD103- mucosal CD8 T cells from CD patients after TCR triggering (n = 11, bars represent means, paired _t_-test, **p < 0.001).

Figure 6

Differential effector functions of CD103 positive and negative mucosal CD8 T cells. (A) Representative expression of CD103, KLRG1 and GZMB on CD8 T cells in blood and ileal mucosa of control individuals (CT) and Crohn's disease (CD) patients. (B) Quantification of GZMB expression on CD8 T cells from blood and ileal mucosa (CT n = 2; CD n = 10; bars represent means; unpaired and paired _t_-test: *p < 0.05) (C) Quantification of GZMB expression on ileal CD103+ and CD103- CD8 T cell subsets (CT n = 2; CD n = 10; bars represent means; paired _t_-test: **p < 0.001). (D) Representative immunostaining of CD103 and GZMB on intestinal mucosa of CD patient. (E) Ratio of CD103 or GZM B positive cells on total mucosal cells (n = 6; bars represent means; paired _t_-test: not significant). (F) Ratio of CD103 or GZM B positive cells on cells in the Lamina propria (LP) or epithelium (EP) (n = 6; bars represent means; paired _t_-test: *p < 0.05, **p < 0.001).

Figure 7

Major transcriptional change between CD103- and CD103+ CD8 Trm cells from CD patient. (A) volcanoplot showing differential expression (fold change >1.5 and P < 0.05) between CD103- and CD103+ T cells subsets from control tissue (CT; _n_ = 3) and CD patients (CD; _n_ = 4) as indicated. **(B)** Venn diagram showing number of gene differentially expressed between CD103- and CD103+ T cells subsets of CT and CD patients and overlapped gene. **(C)** Heatmap of genes differentially expressed (fold change >1.5 and P < 0.05) between CD103- and CD103+ mucosal T cells from CD patients. (D) Quantification and distribution of expression value of genes of interest (dots and bars represent 4 independent experiments with mean and SEM).

Figure 8

CD103+ CD8 Trm cells from CD patient display major transcriptional change compared to CD103+ from CT. (A) Volcanoplot showing differential expression (fold change >2 and P < 0.05) in CD103+ between control (CT; n = 3) and CD patients (CD; n = 4). (B) Heatmap of genes differentially expressed (P < 0.05) between CD103+ mucosal T cells from CT and CD patients. (C) Quantification and distribution of expression value of genes of interest (dots and bars represent 3 and 4 independent experiments with mean and SEM).

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KLRG1 and CD103 Expressions Define Distinct Intestinal Tissue-Resident Memory CD8 T Cell Subsets Modulated in Crohn's Disease - PubMed (original) (raw)