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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

Gut Vγ7+ IELs Respond to Btnl Proteins(A) Left: surface CD25 expression of designated IEL subsets after 12-hr co-culture of total IELs with MODE-K cells transduced with empty vector (EV), Btnl1 (L1), Btnl6 (L6), or Btnl1 plus Btnl6 (L1+6). Right: fold increase in CD25+ cells as percentage of the IEL subset relative to co-culture with MODE-K.EV (n = 21).(B) GFP and CD25 expression by designated IELs from Nur77.gfp mice ex vivo or after 12-hr co-culture of total IELs with designated cells (n = 8).(C) Percentage of Vγ7+ IELs from Nur77.gfp mice that were CD25+GFP+CD122− directly ex vivo or after 12-hr co-culture of total IELs with designated cells.(D) CD3 expression in Vγ7+CD25+ or Vγ7+CD25− IELs after 12-hr co-culture of total IELs with MODE-K.L1+6 (n = 21).(E) Surface CD25 expression as percentage of Vγ7+ IELs after indicated transwell co-culture conditions (n = 3).(F) Surface CD25 expression on Vγ7+ IELs from WT and Btnl1−/− mice after indicated culture conditions (n = 7).(G) Surface CD25 expression by designated IEL subsets after incubation with anti-CD3 or control immunoglobulin (Ig) (n ≥ 12).(H) Cytokine concentrations assessed by luminex in supernatants after 48 hr of co-cultures indicated (n = 3).Data are representative of one (H), two (B) or more than three (A, D, and E) independent experiments. Some panels present data pooled from two (C) or more than three (A, F, and G) independent experiments. All error bars represent mean ± SD. See also Figure S6.
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fig6: Gut Vγ7+ IELs Respond to Btnl Proteins(A) Left: surface CD25 expression of designated IEL subsets after 12-hr co-culture of total IELs with MODE-K cells transduced with empty vector (EV), Btnl1 (L1), Btnl6 (L6), or Btnl1 plus Btnl6 (L1+6). Right: fold increase in CD25+ cells as percentage of the IEL subset relative to co-culture with MODE-K.EV (n = 21).(B) GFP and CD25 expression by designated IELs from Nur77.gfp mice ex vivo or after 12-hr co-culture of total IELs with designated cells (n = 8).(C) Percentage of Vγ7+ IELs from Nur77.gfp mice that were CD25+GFP+CD122− directly ex vivo or after 12-hr co-culture of total IELs with designated cells.(D) CD3 expression in Vγ7+CD25+ or Vγ7+CD25− IELs after 12-hr co-culture of total IELs with MODE-K.L1+6 (n = 21).(E) Surface CD25 expression as percentage of Vγ7+ IELs after indicated transwell co-culture conditions (n = 3).(F) Surface CD25 expression on Vγ7+ IELs from WT and Btnl1−/− mice after indicated culture conditions (n = 7).(G) Surface CD25 expression by designated IEL subsets after incubation with anti-CD3 or control immunoglobulin (Ig) (n ≥ 12).(H) Cytokine concentrations assessed by luminex in supernatants after 48 hr of co-cultures indicated (n = 3).Data are representative of one (H), two (B) or more than three (A, D, and E) independent experiments. Some panels present data pooled from two (C) or more than three (A, F, and G) independent experiments. All error bars represent mean ± SD. See also Figure S6.

Mentions: MODE-K cells stably transduced with Btnl1 (L1), Btnl6 (L6), Btnl1 plus Btnl6 (L1+6), or empty vector (EV) were co-cultured with freshly explanted IELs that were then assayed for CD25 (IL-2Rα chain) upregulation, which is among the most robust readouts of TCR stimulation for systemic T cells (Depper et al., 1984, Kim and Leonard, 2002). Whereas mixed TCRαβ+ and TCRγδ+ IELs showed minor CD25 upregulation upon co-culture with EV, L1, or L6 cells, IELs exposed to L1+6 cells showed highly significant CD25 upregulation, wholly attributable to ∼20% of Vγ7+ cells (both Vγ7GL2+ and Vγ7GL2− cells) (Figure 6A). Significant CD25 upregulation was first evident within 4–6 hr, as is true for systemic TCR stimulation (Depper et al., 1984, Kim and Leonard, 2002) (Figure S6B).


Epithelia Use Butyrophilin-like Molecules to Shape Organ-Specific γ δ T Cell Compartments
Gut Vγ7+ IELs Respond to Btnl Proteins(A) Left: surface CD25 expression of designated IEL subsets after 12-hr co-culture of total IELs with MODE-K cells transduced with empty vector (EV), Btnl1 (L1), Btnl6 (L6), or Btnl1 plus Btnl6 (L1+6). Right: fold increase in CD25+ cells as percentage of the IEL subset relative to co-culture with MODE-K.EV (n = 21).(B) GFP and CD25 expression by designated IELs from Nur77.gfp mice ex vivo or after 12-hr co-culture of total IELs with designated cells (n = 8).(C) Percentage of Vγ7+ IELs from Nur77.gfp mice that were CD25+GFP+CD122− directly ex vivo or after 12-hr co-culture of total IELs with designated cells.(D) CD3 expression in Vγ7+CD25+ or Vγ7+CD25− IELs after 12-hr co-culture of total IELs with MODE-K.L1+6 (n = 21).(E) Surface CD25 expression as percentage of Vγ7+ IELs after indicated transwell co-culture conditions (n = 3).(F) Surface CD25 expression on Vγ7+ IELs from WT and Btnl1−/− mice after indicated culture conditions (n = 7).(G) Surface CD25 expression by designated IEL subsets after incubation with anti-CD3 or control immunoglobulin (Ig) (n ≥ 12).(H) Cytokine concentrations assessed by luminex in supernatants after 48 hr of co-cultures indicated (n = 3).Data are representative of one (H), two (B) or more than three (A, D, and E) independent experiments. Some panels present data pooled from two (C) or more than three (A, F, and G) independent experiments. All error bars represent mean ± SD. See also Figure S6.
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fig6: Gut Vγ7+ IELs Respond to Btnl Proteins(A) Left: surface CD25 expression of designated IEL subsets after 12-hr co-culture of total IELs with MODE-K cells transduced with empty vector (EV), Btnl1 (L1), Btnl6 (L6), or Btnl1 plus Btnl6 (L1+6). Right: fold increase in CD25+ cells as percentage of the IEL subset relative to co-culture with MODE-K.EV (n = 21).(B) GFP and CD25 expression by designated IELs from Nur77.gfp mice ex vivo or after 12-hr co-culture of total IELs with designated cells (n = 8).(C) Percentage of Vγ7+ IELs from Nur77.gfp mice that were CD25+GFP+CD122− directly ex vivo or after 12-hr co-culture of total IELs with designated cells.(D) CD3 expression in Vγ7+CD25+ or Vγ7+CD25− IELs after 12-hr co-culture of total IELs with MODE-K.L1+6 (n = 21).(E) Surface CD25 expression as percentage of Vγ7+ IELs after indicated transwell co-culture conditions (n = 3).(F) Surface CD25 expression on Vγ7+ IELs from WT and Btnl1−/− mice after indicated culture conditions (n = 7).(G) Surface CD25 expression by designated IEL subsets after incubation with anti-CD3 or control immunoglobulin (Ig) (n ≥ 12).(H) Cytokine concentrations assessed by luminex in supernatants after 48 hr of co-cultures indicated (n = 3).Data are representative of one (H), two (B) or more than three (A, D, and E) independent experiments. Some panels present data pooled from two (C) or more than three (A, F, and G) independent experiments. All error bars represent mean ± SD. See also Figure S6.
Mentions: MODE-K cells stably transduced with Btnl1 (L1), Btnl6 (L6), Btnl1 plus Btnl6 (L1+6), or empty vector (EV) were co-cultured with freshly explanted IELs that were then assayed for CD25 (IL-2Rα chain) upregulation, which is among the most robust readouts of TCR stimulation for systemic T cells (Depper et al., 1984, Kim and Leonard, 2002). Whereas mixed TCRαβ+ and TCRγδ+ IELs showed minor CD25 upregulation upon co-culture with EV, L1, or L6 cells, IELs exposed to L1+6 cells showed highly significant CD25 upregulation, wholly attributable to ∼20% of Vγ7+ cells (both Vγ7GL2+ and Vγ7GL2− cells) (Figure 6A). Significant CD25 upregulation was first evident within 4–6 hr, as is true for systemic TCR stimulation (Depper et al., 1984, Kim and Leonard, 2002) (Figure S6B).

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