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Epithelia Use Butyrophilin-like Molecules to Shape Organ-Specific γ δ T Cell Compartments

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ABSTRACT

Many body surfaces harbor organ-specific γδ T cell compartments that contribute to tissue integrity. Thus, murine dendritic epidermal T cells (DETCs) uniquely expressing T cell receptor (TCR)-Vγ5 chains protect from cutaneous carcinogens. The DETC repertoire is shaped by Skint1, a butyrophilin-like (Btnl) gene expressed specifically by thymic epithelial cells and suprabasal keratinocytes. However, the generality of this mechanism has remained opaque, since neither Skint1 nor DETCs are evolutionarily conserved. Here, Btnl1 expressed by murine enterocytes is shown to shape the local TCR-Vγ7+ γδ compartment. Uninfluenced by microbial or food antigens, this activity evokes the developmental selection of TCRαβ+ repertoires. Indeed, Btnl1 and Btnl6 jointly induce TCR-dependent responses specifically in intestinal Vγ7+ cells. Likewise, human gut epithelial cells express BTNL3 and BTNL8 that jointly induce selective TCR-dependent responses of human colonic Vγ4+ cells. Hence, a conserved mechanism emerges whereby epithelia use organ-specific BTNL/Btnl genes to shape local T cell compartments.

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Phenotypic Differences between CD122HI Vγ7+ IELs and Other IEL Subsets, Related to Figure 1(A) Cell surface phenotype of Vγ7+, Vγ7- (CD3+TCRβ-Vγ7-) and αβ (TCRβ+) IEL from 3-5 week old (W3-5) C57Bl/6 (WT) mice (n ≥ 7). (B) Cell surface phenotype of WT Vγ7+CD122HI versus Vγ7+CD122LO IEL (n ≥ 7). (C) Heat map of genes differentially expressed (log-2-FoldChange) between Vγ7+CD122hi and Vγ7+CD122lo IEL sorted from D14-D17 WT mice. Data generated by RNA sequencing (‘cell cycle’ & ‘cell surface’ GO terms annotated). Values scaled to their median value across the samples. (D) 3hr EdU incorporation in vivo in Vγ7+ versus Vγ7- IEL from D28 WT mice assessed by flow cytometry in indicated IEL subsets (Vγ7- are CD3+TCRβ-Vγ7-). Data are representative of 1 (C) or ≥ 3 (A,B) independent experiments. Panel (D) presents data pooled from 3 independent experiments. All error bars represent mean ± SD. Related to Figure 1.
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figs1: Phenotypic Differences between CD122HI Vγ7+ IELs and Other IEL Subsets, Related to Figure 1(A) Cell surface phenotype of Vγ7+, Vγ7- (CD3+TCRβ-Vγ7-) and αβ (TCRβ+) IEL from 3-5 week old (W3-5) C57Bl/6 (WT) mice (n ≥ 7). (B) Cell surface phenotype of WT Vγ7+CD122HI versus Vγ7+CD122LO IEL (n ≥ 7). (C) Heat map of genes differentially expressed (log-2-FoldChange) between Vγ7+CD122hi and Vγ7+CD122lo IEL sorted from D14-D17 WT mice. Data generated by RNA sequencing (‘cell cycle’ & ‘cell surface’ GO terms annotated). Values scaled to their median value across the samples. (D) 3hr EdU incorporation in vivo in Vγ7+ versus Vγ7- IEL from D28 WT mice assessed by flow cytometry in indicated IEL subsets (Vγ7- are CD3+TCRβ-Vγ7-). Data are representative of 1 (C) or ≥ 3 (A,B) independent experiments. Panel (D) presents data pooled from 3 independent experiments. All error bars represent mean ± SD. Related to Figure 1.

Mentions: By flow cytometry of cells recovered from epithelium, and by confocal visualization of epithelial whole mounts, we found that the signature murine small intestinal Vγ7+ IEL compartment largely took shape at 2–3 weeks of age and remained stable for at least 9 months thereafter (Figures 1A and 1B). At day 21, Vγ7+ cells mostly phenocopied mature Skint1-selected DETCs, expressing uniformly high levels of CD122 (the IL-2R/IL-15Rβ chain), TIGIT (an inhibitory co-receptor), and the TCR (detected with anti-CD3 antibodies) and low levels of RNA for Rorgc and Sox13, two transcription factors contributing to γδ T cell differentiation (Vantourout and Hayday, 2013) (Figure 1C). Vγ7− IELs (mostly Vγ1 or Vγ4) did not show this phenotype, and whereas both Vγ7+ and Vγ7− IEL subsets were mostly CD45RBhi, CD44+, and CCR9+, Vγ7+ IELs were distinct in being Lag3+, Thy1−, CD69+, CD5−, and CD8αα+ (Figures 1C and S1A).


Epithelia Use Butyrophilin-like Molecules to Shape Organ-Specific γ δ T Cell Compartments
Phenotypic Differences between CD122HI Vγ7+ IELs and Other IEL Subsets, Related to Figure 1(A) Cell surface phenotype of Vγ7+, Vγ7- (CD3+TCRβ-Vγ7-) and αβ (TCRβ+) IEL from 3-5 week old (W3-5) C57Bl/6 (WT) mice (n ≥ 7). (B) Cell surface phenotype of WT Vγ7+CD122HI versus Vγ7+CD122LO IEL (n ≥ 7). (C) Heat map of genes differentially expressed (log-2-FoldChange) between Vγ7+CD122hi and Vγ7+CD122lo IEL sorted from D14-D17 WT mice. Data generated by RNA sequencing (‘cell cycle’ & ‘cell surface’ GO terms annotated). Values scaled to their median value across the samples. (D) 3hr EdU incorporation in vivo in Vγ7+ versus Vγ7- IEL from D28 WT mice assessed by flow cytometry in indicated IEL subsets (Vγ7- are CD3+TCRβ-Vγ7-). Data are representative of 1 (C) or ≥ 3 (A,B) independent experiments. Panel (D) presents data pooled from 3 independent experiments. All error bars represent mean ± SD. Related to Figure 1.
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figs1: Phenotypic Differences between CD122HI Vγ7+ IELs and Other IEL Subsets, Related to Figure 1(A) Cell surface phenotype of Vγ7+, Vγ7- (CD3+TCRβ-Vγ7-) and αβ (TCRβ+) IEL from 3-5 week old (W3-5) C57Bl/6 (WT) mice (n ≥ 7). (B) Cell surface phenotype of WT Vγ7+CD122HI versus Vγ7+CD122LO IEL (n ≥ 7). (C) Heat map of genes differentially expressed (log-2-FoldChange) between Vγ7+CD122hi and Vγ7+CD122lo IEL sorted from D14-D17 WT mice. Data generated by RNA sequencing (‘cell cycle’ & ‘cell surface’ GO terms annotated). Values scaled to their median value across the samples. (D) 3hr EdU incorporation in vivo in Vγ7+ versus Vγ7- IEL from D28 WT mice assessed by flow cytometry in indicated IEL subsets (Vγ7- are CD3+TCRβ-Vγ7-). Data are representative of 1 (C) or ≥ 3 (A,B) independent experiments. Panel (D) presents data pooled from 3 independent experiments. All error bars represent mean ± SD. Related to Figure 1.
Mentions: By flow cytometry of cells recovered from epithelium, and by confocal visualization of epithelial whole mounts, we found that the signature murine small intestinal Vγ7+ IEL compartment largely took shape at 2–3 weeks of age and remained stable for at least 9 months thereafter (Figures 1A and 1B). At day 21, Vγ7+ cells mostly phenocopied mature Skint1-selected DETCs, expressing uniformly high levels of CD122 (the IL-2R/IL-15Rβ chain), TIGIT (an inhibitory co-receptor), and the TCR (detected with anti-CD3 antibodies) and low levels of RNA for Rorgc and Sox13, two transcription factors contributing to γδ T cell differentiation (Vantourout and Hayday, 2013) (Figure 1C). Vγ7− IELs (mostly Vγ1 or Vγ4) did not show this phenotype, and whereas both Vγ7+ and Vγ7− IEL subsets were mostly CD45RBhi, CD44+, and CCR9+, Vγ7+ IELs were distinct in being Lag3+, Thy1−, CD69+, CD5−, and CD8αα+ (Figures 1C and S1A).

View Article: PubMed Central - PubMed

ABSTRACT

Many body surfaces harbor organ-specific γδ T cell compartments that contribute to tissue integrity. Thus, murine dendritic epidermal T cells (DETCs) uniquely expressing T cell receptor (TCR)-Vγ5 chains protect from cutaneous carcinogens. The DETC repertoire is shaped by Skint1, a butyrophilin-like (Btnl) gene expressed specifically by thymic epithelial cells and suprabasal keratinocytes. However, the generality of this mechanism has remained opaque, since neither Skint1 nor DETCs are evolutionarily conserved. Here, Btnl1 expressed by murine enterocytes is shown to shape the local TCR-Vγ7+ γδ compartment. Uninfluenced by microbial or food antigens, this activity evokes the developmental selection of TCRαβ+ repertoires. Indeed, Btnl1 and Btnl6 jointly induce TCR-dependent responses specifically in intestinal Vγ7+ cells. Likewise, human gut epithelial cells express BTNL3 and BTNL8 that jointly induce selective TCR-dependent responses of human colonic Vγ4+ cells. Hence, a conserved mechanism emerges whereby epithelia use organ-specific BTNL/Btnl genes to shape local T cell compartments.

No MeSH data available.


Related in: MedlinePlus