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Epithelia Use Butyrophilin-like Molecules to Shape Organ-Specific Ī³ Ī“ 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.


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Co-expression of Btnl1 and Btnl6 and Their Impact on VĪ³7+ IELs, Related to FigureĀ 6(A) Cell surface expression of FLAG-Btnl1, HIS-Btnl4 or HA-Btnl6 co-transfected in MODE-K cells. Histogram overlays show the expression of each BTNL after gating on GFP+ cells (numbers in brackets indicate geometric mean fluorescence intensity, gMFI). (B) Primary small intestinal IEL cultured for the indicated times with MODE-K cells transduced with constructs expressing an empty vector (EV) versus Btnl1+Btnl6 (L1+6) (nĀ = 7). (C) Representative plots of cell surface CD122 and CD25 expression on VĪ³7+ cells after the indicated overnight culture conditions (nĀ = 21). (D) Cell surface CD25 expression in positively FACS-sorted VĪ³7+ IEL after overnight co-culture with MODE-K cells expressing EV versus L1+6 (nĀ = 4). (E) Cell surface CD25 expression in primary VĪ³7+ IEL after overnight co-culture with the indicated MODE-K transductants in the presence PP2, PP3 or vehicle. Data are representative of representative of 2 (A,D), or > 5 (C) independent experiments. Some panels (B,E) present data pooled from 2 independent experiments. All error bars represent mean Ā± SD.
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figs6: Co-expression of Btnl1 and Btnl6 and Their Impact on VĪ³7+ IELs, Related to FigureĀ 6(A) Cell surface expression of FLAG-Btnl1, HIS-Btnl4 or HA-Btnl6 co-transfected in MODE-K cells. Histogram overlays show the expression of each BTNL after gating on GFP+ cells (numbers in brackets indicate geometric mean fluorescence intensity, gMFI). (B) Primary small intestinal IEL cultured for the indicated times with MODE-K cells transduced with constructs expressing an empty vector (EV) versus Btnl1+Btnl6 (L1+6) (nĀ = 7). (C) Representative plots of cell surface CD122 and CD25 expression on VĪ³7+ cells after the indicated overnight culture conditions (nĀ = 21). (D) Cell surface CD25 expression in positively FACS-sorted VĪ³7+ IEL after overnight co-culture with MODE-K cells expressing EV versus L1+6 (nĀ = 4). (E) Cell surface CD25 expression in primary VĪ³7+ IEL after overnight co-culture with the indicated MODE-K transductants in the presence PP2, PP3 or vehicle. Data are representative of representative of 2 (A,D), or > 5 (C) independent experiments. Some panels (B,E) present data pooled from 2 independent experiments. All error bars represent mean Ā± SD.

Mentions: Given that acute Btnl1 expression drives the selective maturation of VĪ³7+ IELs inĀ vivo, we tested whether Btnl1 might show specificity for VĪ³7+ IEL exĀ vivo. Since primary intestinal epithelial cellsĀ reportedly harbor Btnl1 in a complex with Btnl6 (Lebrero-FernĆ”ndez etĀ al., 2016), we sought evidence for heterotypic interactions of Btnl proteins. Indeed, cell surface expression of Btnl1 on Btnl1-transfected MODE-K cells (an established intestinal epithelial cell line in which endogenous Btnl genes are negligibly expressed) was greatly enhanced by co-transfection with Btnl4 or Btnl6 (FigureĀ S6A). Likewise, surface Btnl6 expression was greatly enhanced by Btnl1, but there was no evidence for collaboration between Btnl6 and Btnl4 (FigureĀ S6A). Given the specificity of Btnl6 for Btnl1, and given that Btnl4āˆ’/āˆ’ mice showed no IEL phenotype, we focused on Btnl1 and Btnl6.


Epithelia Use Butyrophilin-like Molecules to Shape Organ-Specific Ī³ Ī“ T Cell Compartments
Co-expression of Btnl1 and Btnl6 and Their Impact on VĪ³7+ IELs, Related to FigureĀ 6(A) Cell surface expression of FLAG-Btnl1, HIS-Btnl4 or HA-Btnl6 co-transfected in MODE-K cells. Histogram overlays show the expression of each BTNL after gating on GFP+ cells (numbers in brackets indicate geometric mean fluorescence intensity, gMFI). (B) Primary small intestinal IEL cultured for the indicated times with MODE-K cells transduced with constructs expressing an empty vector (EV) versus Btnl1+Btnl6 (L1+6) (nĀ = 7). (C) Representative plots of cell surface CD122 and CD25 expression on VĪ³7+ cells after the indicated overnight culture conditions (nĀ = 21). (D) Cell surface CD25 expression in positively FACS-sorted VĪ³7+ IEL after overnight co-culture with MODE-K cells expressing EV versus L1+6 (nĀ = 4). (E) Cell surface CD25 expression in primary VĪ³7+ IEL after overnight co-culture with the indicated MODE-K transductants in the presence PP2, PP3 or vehicle. Data are representative of representative of 2 (A,D), or > 5 (C) independent experiments. Some panels (B,E) present data pooled from 2 independent experiments. All error bars represent mean Ā± SD.
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figs6: Co-expression of Btnl1 and Btnl6 and Their Impact on VĪ³7+ IELs, Related to FigureĀ 6(A) Cell surface expression of FLAG-Btnl1, HIS-Btnl4 or HA-Btnl6 co-transfected in MODE-K cells. Histogram overlays show the expression of each BTNL after gating on GFP+ cells (numbers in brackets indicate geometric mean fluorescence intensity, gMFI). (B) Primary small intestinal IEL cultured for the indicated times with MODE-K cells transduced with constructs expressing an empty vector (EV) versus Btnl1+Btnl6 (L1+6) (nĀ = 7). (C) Representative plots of cell surface CD122 and CD25 expression on VĪ³7+ cells after the indicated overnight culture conditions (nĀ = 21). (D) Cell surface CD25 expression in positively FACS-sorted VĪ³7+ IEL after overnight co-culture with MODE-K cells expressing EV versus L1+6 (nĀ = 4). (E) Cell surface CD25 expression in primary VĪ³7+ IEL after overnight co-culture with the indicated MODE-K transductants in the presence PP2, PP3 or vehicle. Data are representative of representative of 2 (A,D), or > 5 (C) independent experiments. Some panels (B,E) present data pooled from 2 independent experiments. All error bars represent mean Ā± SD.
Mentions: Given that acute Btnl1 expression drives the selective maturation of VĪ³7+ IELs inĀ vivo, we tested whether Btnl1 might show specificity for VĪ³7+ IEL exĀ vivo. Since primary intestinal epithelial cellsĀ reportedly harbor Btnl1 in a complex with Btnl6 (Lebrero-FernĆ”ndez etĀ al., 2016), we sought evidence for heterotypic interactions of Btnl proteins. Indeed, cell surface expression of Btnl1 on Btnl1-transfected MODE-K cells (an established intestinal epithelial cell line in which endogenous Btnl genes are negligibly expressed) was greatly enhanced by co-transfection with Btnl4 or Btnl6 (FigureĀ S6A). Likewise, surface Btnl6 expression was greatly enhanced by Btnl1, but there was no evidence for collaboration between Btnl6 and Btnl4 (FigureĀ S6A). Given the specificity of Btnl6 for Btnl1, and given that Btnl4āˆ’/āˆ’ mice showed no IEL phenotype, we focused on Btnl1 and Btnl6.

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