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Linear ubiquitin chain assembly complex coordinates late thymic T-cell differentiation and regulatory T-cell homeostasis

View Article: PubMed Central - PubMed

ABSTRACT

The linear ubiquitin chain assembly complex (LUBAC) is essential for innate immunity in mice and humans, yet its role in adaptive immunity is unclear. Here we show that the LUBAC components HOIP, HOIL-1 and SHARPIN have essential roles in late thymocyte differentiation, FOXP3+ regulatory T (Treg)-cell development and Treg cell homeostasis. LUBAC activity is not required to prevent TNF-induced apoptosis or necroptosis but is necessary for the transcriptional programme of the penultimate stage of thymocyte differentiation. Treg cell-specific ablation of HOIP causes severe Treg cell deficiency and lethal immune pathology, revealing an ongoing requirement of LUBAC activity for Treg cell homeostasis. These data reveal stage-specific requirements for LUBAC in coordinating the signals required for T-cell differentiation.

No MeSH data available.


Related in: MedlinePlus

Absence of HOIP or HOIL-1 causes T-cell deficiency.(a) Flow cytometry of splenic cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm assessed for expression of TCRβ (top panels), CD4 and CD8 (middle panels), and for FOXP3, after gating on CD4+ cells (bottom panels). (b) Quantification of CD4+CD8+TCRβ+ and CD4+FOXP3+ cells in the spleen. (c) Surface expression of CD44 and CD62L on CD4+FOXP3− (top panels) and CD8+ T cells (bottom panels) in spleens of control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (d) Absolute numbers of CD44highCD62Llow activated cells in the CD4+FOXP3− (left graph) and CD8+ (right graph) populations from the spleens of controls, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (e) Flow cytometry of splenic CD1d-αGalCer tetramer-positive NKT cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (f) Total cell numbers and percentages of CD1d-α-galactosylceramide (α-GalCer) tetramer-positive NKT cells in the spleen. For b,d and f, each symbol represents an individual mouse; small horizontal lines indicate mean±s.d.; *P<0.05, **P<0.01, ***P<0.005 and ****P<0.001, respectively. One-way analysis of variance with a Tukey's post-hoc test for multiple comparisons was used for statistical analysis. Shpncpdm refers to Sharpincpdm mice. Data are pooled from six independent experiments with two to six mice per group (a–d) or representative of two independent experiments with three to six mice per group (e,f).
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f1: Absence of HOIP or HOIL-1 causes T-cell deficiency.(a) Flow cytometry of splenic cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm assessed for expression of TCRβ (top panels), CD4 and CD8 (middle panels), and for FOXP3, after gating on CD4+ cells (bottom panels). (b) Quantification of CD4+CD8+TCRβ+ and CD4+FOXP3+ cells in the spleen. (c) Surface expression of CD44 and CD62L on CD4+FOXP3− (top panels) and CD8+ T cells (bottom panels) in spleens of control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (d) Absolute numbers of CD44highCD62Llow activated cells in the CD4+FOXP3− (left graph) and CD8+ (right graph) populations from the spleens of controls, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (e) Flow cytometry of splenic CD1d-αGalCer tetramer-positive NKT cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (f) Total cell numbers and percentages of CD1d-α-galactosylceramide (α-GalCer) tetramer-positive NKT cells in the spleen. For b,d and f, each symbol represents an individual mouse; small horizontal lines indicate mean±s.d.; *P<0.05, **P<0.01, ***P<0.005 and ****P<0.001, respectively. One-way analysis of variance with a Tukey's post-hoc test for multiple comparisons was used for statistical analysis. Shpncpdm refers to Sharpincpdm mice. Data are pooled from six independent experiments with two to six mice per group (a–d) or representative of two independent experiments with three to six mice per group (e,f).

Mentions: The peripheral immune organs of HoipΔCd4 and HoilΔCd4 mice were almost completely devoid of CD8+ and CD4+ αβTCR+ T cells (Fig. 1a,b). Although the proportions of FOXP3+ Treg cells among αβTCR+ CD4+ T cells were normal in HoipΔCd4 and HoilΔCd4 mice, their number was greatly reduced, in line with the overall T-cell deficiency (Fig. 1a,b). The residual αβTCR+ T cells in HoipΔCd4 and HoilΔCd4 mice were predominantly CD44hi CD62Llo (Fig. 1c,d), suggestive of an activated/effector phenotype that is often associated with ‘homeostatic' expansion during lymphopenia24. By contrast, the numbers, proportions and activation status of conventional CD8+ and CD4+ T cells in Sharpincpdm mice were comparable to controls (Fig. 1a–d). Consistent with recent reports2526, we observed that Treg cells were reduced in Sharpincpdm mice, although to a much lesser extent compared with the HoipΔCd4 and HoilΔCd4 mice (Fig. 1a,b). These data indicate that the LUBAC components HOIL-1 and HOIP are necessary for conventional CD4+ and CD8+ T-cell differentiation, and that SHARPIN is dispensable.


Linear ubiquitin chain assembly complex coordinates late thymic T-cell differentiation and regulatory T-cell homeostasis
Absence of HOIP or HOIL-1 causes T-cell deficiency.(a) Flow cytometry of splenic cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm assessed for expression of TCRβ (top panels), CD4 and CD8 (middle panels), and for FOXP3, after gating on CD4+ cells (bottom panels). (b) Quantification of CD4+CD8+TCRβ+ and CD4+FOXP3+ cells in the spleen. (c) Surface expression of CD44 and CD62L on CD4+FOXP3− (top panels) and CD8+ T cells (bottom panels) in spleens of control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (d) Absolute numbers of CD44highCD62Llow activated cells in the CD4+FOXP3− (left graph) and CD8+ (right graph) populations from the spleens of controls, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (e) Flow cytometry of splenic CD1d-αGalCer tetramer-positive NKT cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (f) Total cell numbers and percentages of CD1d-α-galactosylceramide (α-GalCer) tetramer-positive NKT cells in the spleen. For b,d and f, each symbol represents an individual mouse; small horizontal lines indicate mean±s.d.; *P<0.05, **P<0.01, ***P<0.005 and ****P<0.001, respectively. One-way analysis of variance with a Tukey's post-hoc test for multiple comparisons was used for statistical analysis. Shpncpdm refers to Sharpincpdm mice. Data are pooled from six independent experiments with two to six mice per group (a–d) or representative of two independent experiments with three to six mice per group (e,f).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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f1: Absence of HOIP or HOIL-1 causes T-cell deficiency.(a) Flow cytometry of splenic cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm assessed for expression of TCRβ (top panels), CD4 and CD8 (middle panels), and for FOXP3, after gating on CD4+ cells (bottom panels). (b) Quantification of CD4+CD8+TCRβ+ and CD4+FOXP3+ cells in the spleen. (c) Surface expression of CD44 and CD62L on CD4+FOXP3− (top panels) and CD8+ T cells (bottom panels) in spleens of control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (d) Absolute numbers of CD44highCD62Llow activated cells in the CD4+FOXP3− (left graph) and CD8+ (right graph) populations from the spleens of controls, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (e) Flow cytometry of splenic CD1d-αGalCer tetramer-positive NKT cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (f) Total cell numbers and percentages of CD1d-α-galactosylceramide (α-GalCer) tetramer-positive NKT cells in the spleen. For b,d and f, each symbol represents an individual mouse; small horizontal lines indicate mean±s.d.; *P<0.05, **P<0.01, ***P<0.005 and ****P<0.001, respectively. One-way analysis of variance with a Tukey's post-hoc test for multiple comparisons was used for statistical analysis. Shpncpdm refers to Sharpincpdm mice. Data are pooled from six independent experiments with two to six mice per group (a–d) or representative of two independent experiments with three to six mice per group (e,f).
Mentions: The peripheral immune organs of HoipΔCd4 and HoilΔCd4 mice were almost completely devoid of CD8+ and CD4+ αβTCR+ T cells (Fig. 1a,b). Although the proportions of FOXP3+ Treg cells among αβTCR+ CD4+ T cells were normal in HoipΔCd4 and HoilΔCd4 mice, their number was greatly reduced, in line with the overall T-cell deficiency (Fig. 1a,b). The residual αβTCR+ T cells in HoipΔCd4 and HoilΔCd4 mice were predominantly CD44hi CD62Llo (Fig. 1c,d), suggestive of an activated/effector phenotype that is often associated with ‘homeostatic' expansion during lymphopenia24. By contrast, the numbers, proportions and activation status of conventional CD8+ and CD4+ T cells in Sharpincpdm mice were comparable to controls (Fig. 1a–d). Consistent with recent reports2526, we observed that Treg cells were reduced in Sharpincpdm mice, although to a much lesser extent compared with the HoipΔCd4 and HoilΔCd4 mice (Fig. 1a,b). These data indicate that the LUBAC components HOIL-1 and HOIP are necessary for conventional CD4+ and CD8+ T-cell differentiation, and that SHARPIN is dispensable.

View Article: PubMed Central - PubMed

ABSTRACT

The linear ubiquitin chain assembly complex (LUBAC) is essential for innate immunity in mice and humans, yet its role in adaptive immunity is unclear. Here we show that the LUBAC components HOIP, HOIL-1 and SHARPIN have essential roles in late thymocyte differentiation, FOXP3+ regulatory T (Treg)-cell development and Treg cell homeostasis. LUBAC activity is not required to prevent TNF-induced apoptosis or necroptosis but is necessary for the transcriptional programme of the penultimate stage of thymocyte differentiation. Treg cell-specific ablation of HOIP causes severe Treg cell deficiency and lethal immune pathology, revealing an ongoing requirement of LUBAC activity for Treg cell homeostasis. These data reveal stage-specific requirements for LUBAC in coordinating the signals required for T-cell differentiation.

No MeSH data available.


Related in: MedlinePlus