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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.


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Local Intestinal Development of CD122HI Vγ7+ IELs, Related to Figure 2(A) Deep sequencing of TCR Vδ chain usage in WT Vγ7+ IEL sorted from W7-10 C57Bl/6 (WT) mice (n = 3). (B) Absolute numbers of WT Vγ7+ and Vγ7+GL2+ thymocytes from WT mice assessed by flow cytometry. C-D) Cell surface phenotype of WT Vγ7+ IEL and thymocytes at indicated time points. (E) Cell surface phenotype of Vγ7+ thymocytes and IEL isolated from W3-5 WT mice (n = 5). (F) γδ IEL composition (left), cell count (middle) and cell surface CD122 expression (right) in WT versus alymphoplasia (aly/aly) mice. (G) Longitudinal RNAscope analysis of Btnl1 expression during gut development. (H) Gene expression by qRT.PCR along the length of the gut in WT mice (n ≥ 3). (I) Gene expression by qRT.PCR in the thymus of WT and Btnl1−/− animals compared to the proximal small intestine. (J) Organization of WT and targeted loci for Btnl1−/−, Btnl1indel/indel and Btnl4−/− mice. Grey: untranslated region; green: translated region; orange: inserted targeting cassette. Knockout ES cell clones were obtained from the international mouse consortium IKMC-ID 67994 (Btnl1) and 81524 (Btnl4). (K) Southern blot for targeting of alleles in Btnl1−/− and Btnl4−/− mice. Genomic DNA was digested using the indicated enzymes (arrowheads). Probes targeting the indicated regions were generated to detect the WT and targeted alleles. Data are representative of ≥ 1 (A,K) or ≥ 2 (C,E,G,H,I) independent experiments. Some panels include data pooled from 2 (F), > 3 (D) or > 6 (B) independent experiments. All error bars represent mean ± SD. Related to Figure 2
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figs2: Local Intestinal Development of CD122HI Vγ7+ IELs, Related to Figure 2(A) Deep sequencing of TCR Vδ chain usage in WT Vγ7+ IEL sorted from W7-10 C57Bl/6 (WT) mice (n = 3). (B) Absolute numbers of WT Vγ7+ and Vγ7+GL2+ thymocytes from WT mice assessed by flow cytometry. C-D) Cell surface phenotype of WT Vγ7+ IEL and thymocytes at indicated time points. (E) Cell surface phenotype of Vγ7+ thymocytes and IEL isolated from W3-5 WT mice (n = 5). (F) γδ IEL composition (left), cell count (middle) and cell surface CD122 expression (right) in WT versus alymphoplasia (aly/aly) mice. (G) Longitudinal RNAscope analysis of Btnl1 expression during gut development. (H) Gene expression by qRT.PCR along the length of the gut in WT mice (n ≥ 3). (I) Gene expression by qRT.PCR in the thymus of WT and Btnl1−/− animals compared to the proximal small intestine. (J) Organization of WT and targeted loci for Btnl1−/−, Btnl1indel/indel and Btnl4−/− mice. Grey: untranslated region; green: translated region; orange: inserted targeting cassette. Knockout ES cell clones were obtained from the international mouse consortium IKMC-ID 67994 (Btnl1) and 81524 (Btnl4). (K) Southern blot for targeting of alleles in Btnl1−/− and Btnl4−/− mice. Genomic DNA was digested using the indicated enzymes (arrowheads). Probes targeting the indicated regions were generated to detect the WT and targeted alleles. Data are representative of ≥ 1 (A,K) or ≥ 2 (C,E,G,H,I) independent experiments. Some panels include data pooled from 2 (F), > 3 (D) or > 6 (B) independent experiments. All error bars represent mean ± SD. Related to Figure 2

Mentions: Because Skint1 selects for signature Vγ5+ DETC progenitors in the thymus, DETCs are absent from athymic NU/NU mice. By contrast, intestinal IELs were present in NU/NU, and although there was some decrease in numbers (average of ∼1.3 × 106 cells compared to >2.0 × 106 cells in euthymic mice; see below), the compartment was again dominated by CD122hi Vγ7+ IELs. Moreover, ∼25% of Vγ7+ IELs in NU/NU and in euthymic mice reacted with antibody GL2 that detects Vδ4 (TRDV2-2 encoded) chains. Consistent with this, TRDV2-2 sequences accounted for ∼25% of TCRδ chain RNAs expressed by purified Vγ7+ IELs (Figures 2A and S2A). In sum, the shaping of the gut Vγ7+ IEL compartment did not require a thymus.


Epithelia Use Butyrophilin-like Molecules to Shape Organ-Specific γ δ T Cell Compartments
Local Intestinal Development of CD122HI Vγ7+ IELs, Related to Figure 2(A) Deep sequencing of TCR Vδ chain usage in WT Vγ7+ IEL sorted from W7-10 C57Bl/6 (WT) mice (n = 3). (B) Absolute numbers of WT Vγ7+ and Vγ7+GL2+ thymocytes from WT mice assessed by flow cytometry. C-D) Cell surface phenotype of WT Vγ7+ IEL and thymocytes at indicated time points. (E) Cell surface phenotype of Vγ7+ thymocytes and IEL isolated from W3-5 WT mice (n = 5). (F) γδ IEL composition (left), cell count (middle) and cell surface CD122 expression (right) in WT versus alymphoplasia (aly/aly) mice. (G) Longitudinal RNAscope analysis of Btnl1 expression during gut development. (H) Gene expression by qRT.PCR along the length of the gut in WT mice (n ≥ 3). (I) Gene expression by qRT.PCR in the thymus of WT and Btnl1−/− animals compared to the proximal small intestine. (J) Organization of WT and targeted loci for Btnl1−/−, Btnl1indel/indel and Btnl4−/− mice. Grey: untranslated region; green: translated region; orange: inserted targeting cassette. Knockout ES cell clones were obtained from the international mouse consortium IKMC-ID 67994 (Btnl1) and 81524 (Btnl4). (K) Southern blot for targeting of alleles in Btnl1−/− and Btnl4−/− mice. Genomic DNA was digested using the indicated enzymes (arrowheads). Probes targeting the indicated regions were generated to detect the WT and targeted alleles. Data are representative of ≥ 1 (A,K) or ≥ 2 (C,E,G,H,I) independent experiments. Some panels include data pooled from 2 (F), > 3 (D) or > 6 (B) independent experiments. All error bars represent mean ± SD. Related to Figure 2
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figs2: Local Intestinal Development of CD122HI Vγ7+ IELs, Related to Figure 2(A) Deep sequencing of TCR Vδ chain usage in WT Vγ7+ IEL sorted from W7-10 C57Bl/6 (WT) mice (n = 3). (B) Absolute numbers of WT Vγ7+ and Vγ7+GL2+ thymocytes from WT mice assessed by flow cytometry. C-D) Cell surface phenotype of WT Vγ7+ IEL and thymocytes at indicated time points. (E) Cell surface phenotype of Vγ7+ thymocytes and IEL isolated from W3-5 WT mice (n = 5). (F) γδ IEL composition (left), cell count (middle) and cell surface CD122 expression (right) in WT versus alymphoplasia (aly/aly) mice. (G) Longitudinal RNAscope analysis of Btnl1 expression during gut development. (H) Gene expression by qRT.PCR along the length of the gut in WT mice (n ≥ 3). (I) Gene expression by qRT.PCR in the thymus of WT and Btnl1−/− animals compared to the proximal small intestine. (J) Organization of WT and targeted loci for Btnl1−/−, Btnl1indel/indel and Btnl4−/− mice. Grey: untranslated region; green: translated region; orange: inserted targeting cassette. Knockout ES cell clones were obtained from the international mouse consortium IKMC-ID 67994 (Btnl1) and 81524 (Btnl4). (K) Southern blot for targeting of alleles in Btnl1−/− and Btnl4−/− mice. Genomic DNA was digested using the indicated enzymes (arrowheads). Probes targeting the indicated regions were generated to detect the WT and targeted alleles. Data are representative of ≥ 1 (A,K) or ≥ 2 (C,E,G,H,I) independent experiments. Some panels include data pooled from 2 (F), > 3 (D) or > 6 (B) independent experiments. All error bars represent mean ± SD. Related to Figure 2
Mentions: Because Skint1 selects for signature Vγ5+ DETC progenitors in the thymus, DETCs are absent from athymic NU/NU mice. By contrast, intestinal IELs were present in NU/NU, and although there was some decrease in numbers (average of ∼1.3 × 106 cells compared to >2.0 × 106 cells in euthymic mice; see below), the compartment was again dominated by CD122hi Vγ7+ IELs. Moreover, ∼25% of Vγ7+ IELs in NU/NU and in euthymic mice reacted with antibody GL2 that detects Vδ4 (TRDV2-2 encoded) chains. Consistent with this, TRDV2-2 sequences accounted for ∼25% of TCRδ chain RNAs expressed by purified Vγ7+ IELs (Figures 2A and S2A). In sum, the shaping of the gut Vγ7+ IEL compartment did not require a thymus.

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