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

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

Selective Maturation and Expansion of Intestinal IELs in Mice(A) Gating strategy for small intestinal (SI) Vγ7+ IELs in 12-week-old C57Bl/6 mice (n ≥ 12). Bottom right: Vγ7+ IEL representation over time (n = 5, week 20–36; n ≥ 12, other time points).(B) IEL composition assessed by confocal microscopy of proximal SI whole mounts (n = 3) and corresponding quantification (right).(C) Top: surface phenotypes of Vγ7+, Vγ7− (CD3+TCRβ−Vγ7−) and αβ IELs from 21- to 40-day-old mice (n ≥ 8). Bottom: gene expression in Vγ7+ versus Vγ7− IELs (n = 3).(D) Surface phenotypes of Vγ7+ IELs at days 14–17 versus days 21–40 (n ≥ 7).(E) Top: surface phenotype of Vγ7+ IELs at day 14 and day 28 (CD122 median fluorescence intensity [MFI]-colored text). Bottom: surface phenotype of CD122HIThy1−Vγ7+ versus CD122LOThy1+Vγ7+ IELs at days 14–17 (n ≥ 7).(F) Heatmap of genes differentially expressed between Vγ7+CD122HI and Vγ7+CD122LO IELs from day 14–17 mice and between Skint1-selected and non-selected Vγ5+ DETC progenitors (n = 4).(G) Ki67 expression in Vγ7+ versus Vγ7− IELs directly ex vivo (n = 4, day 19; n = 8–27, other time points).Data are representative of one (C, qPCR, B and F) or three or more (C, cytometry, D and E, top) independent experiments. Some panels present data pooled from three or more (E), more than ten (G), and >20 (A) independent experiments. D, day; W, week.All error bars represent mean ± SD. See also Figure S1.
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fig1: Selective Maturation and Expansion of Intestinal IELs in Mice(A) Gating strategy for small intestinal (SI) Vγ7+ IELs in 12-week-old C57Bl/6 mice (n ≥ 12). Bottom right: Vγ7+ IEL representation over time (n = 5, week 20–36; n ≥ 12, other time points).(B) IEL composition assessed by confocal microscopy of proximal SI whole mounts (n = 3) and corresponding quantification (right).(C) Top: surface phenotypes of Vγ7+, Vγ7− (CD3+TCRβ−Vγ7−) and αβ IELs from 21- to 40-day-old mice (n ≥ 8). Bottom: gene expression in Vγ7+ versus Vγ7− IELs (n = 3).(D) Surface phenotypes of Vγ7+ IELs at days 14–17 versus days 21–40 (n ≥ 7).(E) Top: surface phenotype of Vγ7+ IELs at day 14 and day 28 (CD122 median fluorescence intensity [MFI]-colored text). Bottom: surface phenotype of CD122HIThy1−Vγ7+ versus CD122LOThy1+Vγ7+ IELs at days 14–17 (n ≥ 7).(F) Heatmap of genes differentially expressed between Vγ7+CD122HI and Vγ7+CD122LO IELs from day 14–17 mice and between Skint1-selected and non-selected Vγ5+ DETC progenitors (n = 4).(G) Ki67 expression in Vγ7+ versus Vγ7− IELs directly ex vivo (n = 4, day 19; n = 8–27, other time points).Data are representative of one (C, qPCR, B and F) or three or more (C, cytometry, D and E, top) independent experiments. Some panels present data pooled from three or more (E), more than ten (G), and >20 (A) independent experiments. D, day; W, week.All error bars represent mean ± SD. See also Figure S1.

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
Selective Maturation and Expansion of Intestinal IELs in Mice(A) Gating strategy for small intestinal (SI) Vγ7+ IELs in 12-week-old C57Bl/6 mice (n ≥ 12). Bottom right: Vγ7+ IEL representation over time (n = 5, week 20–36; n ≥ 12, other time points).(B) IEL composition assessed by confocal microscopy of proximal SI whole mounts (n = 3) and corresponding quantification (right).(C) Top: surface phenotypes of Vγ7+, Vγ7− (CD3+TCRβ−Vγ7−) and αβ IELs from 21- to 40-day-old mice (n ≥ 8). Bottom: gene expression in Vγ7+ versus Vγ7− IELs (n = 3).(D) Surface phenotypes of Vγ7+ IELs at days 14–17 versus days 21–40 (n ≥ 7).(E) Top: surface phenotype of Vγ7+ IELs at day 14 and day 28 (CD122 median fluorescence intensity [MFI]-colored text). Bottom: surface phenotype of CD122HIThy1−Vγ7+ versus CD122LOThy1+Vγ7+ IELs at days 14–17 (n ≥ 7).(F) Heatmap of genes differentially expressed between Vγ7+CD122HI and Vγ7+CD122LO IELs from day 14–17 mice and between Skint1-selected and non-selected Vγ5+ DETC progenitors (n = 4).(G) Ki67 expression in Vγ7+ versus Vγ7− IELs directly ex vivo (n = 4, day 19; n = 8–27, other time points).Data are representative of one (C, qPCR, B and F) or three or more (C, cytometry, D and E, top) independent experiments. Some panels present data pooled from three or more (E), more than ten (G), and >20 (A) independent experiments. D, day; W, week.All error bars represent mean ± SD. See also Figure S1.
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fig1: Selective Maturation and Expansion of Intestinal IELs in Mice(A) Gating strategy for small intestinal (SI) Vγ7+ IELs in 12-week-old C57Bl/6 mice (n ≥ 12). Bottom right: Vγ7+ IEL representation over time (n = 5, week 20–36; n ≥ 12, other time points).(B) IEL composition assessed by confocal microscopy of proximal SI whole mounts (n = 3) and corresponding quantification (right).(C) Top: surface phenotypes of Vγ7+, Vγ7− (CD3+TCRβ−Vγ7−) and αβ IELs from 21- to 40-day-old mice (n ≥ 8). Bottom: gene expression in Vγ7+ versus Vγ7− IELs (n = 3).(D) Surface phenotypes of Vγ7+ IELs at days 14–17 versus days 21–40 (n ≥ 7).(E) Top: surface phenotype of Vγ7+ IELs at day 14 and day 28 (CD122 median fluorescence intensity [MFI]-colored text). Bottom: surface phenotype of CD122HIThy1−Vγ7+ versus CD122LOThy1+Vγ7+ IELs at days 14–17 (n ≥ 7).(F) Heatmap of genes differentially expressed between Vγ7+CD122HI and Vγ7+CD122LO IELs from day 14–17 mice and between Skint1-selected and non-selected Vγ5+ DETC progenitors (n = 4).(G) Ki67 expression in Vγ7+ versus Vγ7− IELs directly ex vivo (n = 4, day 19; n = 8–27, other time points).Data are representative of one (C, qPCR, B and F) or three or more (C, cytometry, D and E, top) independent experiments. Some panels present data pooled from three or more (E), more than ten (G), and >20 (A) independent experiments. D, day; W, week.All error bars represent mean ± SD. See also Figure S1.
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