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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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Regulation of Human Gut Vγ4+ Cells by BTNL3 and BTNL8(A and B) Vδ (A, n = 17) and Vγ (B, n = 6–10) expression by human gut γδ cells.(C) Surface TCRγδ/Vδ1 expression on human gut lymphocytes after 12-hr co-culture with BTNL3 (L3) or BTNL3 plus BTNL8 (L3+8)-transduced HEK293T cells. Red arrows denote shifts in TCR staining.(D) Top: TCRγδ/CD3 expression on designated human gut T cells after 12-hr co-culture with denoted HEK293T transductants. Bottom: mean fluorescence intensities (MFIs) calculated relative to co-culture with HEK293T.EV (n ≥ 22). For two donors, MFIs for Vδ2− cells remained unchanged (dots within the ellipse).(E) Percentage of CD25HI cells among TCRγδ+TCRVδ2− T cells after co-culture with denoted cells (n = 5). Statistical analysis was performed by paired t test.(F) Surface Vγ2/3/4 and Vδ1 expression on Vδ2− γδ T cells after co-culture with cells denoted.(G) TCRγδ expression on indicated subsets after co-culture with denoted cells.(H) TCRγδ expression on γδ cells from peripheral blood mononuclear cells (PBMCs) or skin after co-culture with denoted cells.All error bars represent mean ± SD. See also Figure S7 and Table S1.
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fig7: Regulation of Human Gut Vγ4+ Cells by BTNL3 and BTNL8(A and B) Vδ (A, n = 17) and Vγ (B, n = 6–10) expression by human gut γδ cells.(C) Surface TCRγδ/Vδ1 expression on human gut lymphocytes after 12-hr co-culture with BTNL3 (L3) or BTNL3 plus BTNL8 (L3+8)-transduced HEK293T cells. Red arrows denote shifts in TCR staining.(D) Top: TCRγδ/CD3 expression on designated human gut T cells after 12-hr co-culture with denoted HEK293T transductants. Bottom: mean fluorescence intensities (MFIs) calculated relative to co-culture with HEK293T.EV (n ≥ 22). For two donors, MFIs for Vδ2− cells remained unchanged (dots within the ellipse).(E) Percentage of CD25HI cells among TCRγδ+TCRVδ2− T cells after co-culture with denoted cells (n = 5). Statistical analysis was performed by paired t test.(F) Surface Vγ2/3/4 and Vδ1 expression on Vδ2− γδ T cells after co-culture with cells denoted.(G) TCRγδ expression on indicated subsets after co-culture with denoted cells.(H) TCRγδ expression on γδ cells from peripheral blood mononuclear cells (PBMCs) or skin after co-culture with denoted cells.All error bars represent mean ± SD. See also Figure S7 and Table S1.

Mentions: Because of limited tissue access, human gut T cells are understudied. Nonetheless, there are gut-associated γδ cells whose TCR usage differs markedly from Vγ9+Vδ2+ cells that dominate the peripheral blood (Landau et al., 1995). To better characterize such cells, we submitted biopsy specimens from healthy ascending colon to a modified version of a protocol used to isolate human skin T cells (Clark et al., 2006). For 16 of 17 donors, the γδ cells were enriched in Vδ1+ cells, although Vδ1−Vδ2− cells were also present; hence, the term “Vδ2−” is used to distinguish tissue-associated γδ cells from Vδ2+ cells that could also be recovered from most gut samples, albeit in highly variable numbers (Figure 7A).


Epithelia Use Butyrophilin-like Molecules to Shape Organ-Specific γ δ T Cell Compartments
Regulation of Human Gut Vγ4+ Cells by BTNL3 and BTNL8(A and B) Vδ (A, n = 17) and Vγ (B, n = 6–10) expression by human gut γδ cells.(C) Surface TCRγδ/Vδ1 expression on human gut lymphocytes after 12-hr co-culture with BTNL3 (L3) or BTNL3 plus BTNL8 (L3+8)-transduced HEK293T cells. Red arrows denote shifts in TCR staining.(D) Top: TCRγδ/CD3 expression on designated human gut T cells after 12-hr co-culture with denoted HEK293T transductants. Bottom: mean fluorescence intensities (MFIs) calculated relative to co-culture with HEK293T.EV (n ≥ 22). For two donors, MFIs for Vδ2− cells remained unchanged (dots within the ellipse).(E) Percentage of CD25HI cells among TCRγδ+TCRVδ2− T cells after co-culture with denoted cells (n = 5). Statistical analysis was performed by paired t test.(F) Surface Vγ2/3/4 and Vδ1 expression on Vδ2− γδ T cells after co-culture with cells denoted.(G) TCRγδ expression on indicated subsets after co-culture with denoted cells.(H) TCRγδ expression on γδ cells from peripheral blood mononuclear cells (PBMCs) or skin after co-culture with denoted cells.All error bars represent mean ± SD. See also Figure S7 and Table S1.
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fig7: Regulation of Human Gut Vγ4+ Cells by BTNL3 and BTNL8(A and B) Vδ (A, n = 17) and Vγ (B, n = 6–10) expression by human gut γδ cells.(C) Surface TCRγδ/Vδ1 expression on human gut lymphocytes after 12-hr co-culture with BTNL3 (L3) or BTNL3 plus BTNL8 (L3+8)-transduced HEK293T cells. Red arrows denote shifts in TCR staining.(D) Top: TCRγδ/CD3 expression on designated human gut T cells after 12-hr co-culture with denoted HEK293T transductants. Bottom: mean fluorescence intensities (MFIs) calculated relative to co-culture with HEK293T.EV (n ≥ 22). For two donors, MFIs for Vδ2− cells remained unchanged (dots within the ellipse).(E) Percentage of CD25HI cells among TCRγδ+TCRVδ2− T cells after co-culture with denoted cells (n = 5). Statistical analysis was performed by paired t test.(F) Surface Vγ2/3/4 and Vδ1 expression on Vδ2− γδ T cells after co-culture with cells denoted.(G) TCRγδ expression on indicated subsets after co-culture with denoted cells.(H) TCRγδ expression on γδ cells from peripheral blood mononuclear cells (PBMCs) or skin after co-culture with denoted cells.All error bars represent mean ± SD. See also Figure S7 and Table S1.
Mentions: Because of limited tissue access, human gut T cells are understudied. Nonetheless, there are gut-associated γδ cells whose TCR usage differs markedly from Vγ9+Vδ2+ cells that dominate the peripheral blood (Landau et al., 1995). To better characterize such cells, we submitted biopsy specimens from healthy ascending colon to a modified version of a protocol used to isolate human skin T cells (Clark et al., 2006). For 16 of 17 donors, the γδ cells were enriched in Vδ1+ cells, although Vδ1−Vδ2− cells were also present; hence, the term “Vδ2−” is used to distinguish tissue-associated γδ cells from Vδ2+ cells that could also be recovered from most gut samples, albeit in highly variable numbers (Figure 7A).

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