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

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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.


Thymic T-cell differentiation in LUBAC-deficient mice.(a) Surface staining of CD4 and CD8 (upper panels), and FOXP3 and CD25 (lower panels) on thymocytes from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (b) Quantification of total cell numbers and percentages of CD4+ CD8+ and FOXP3+ thymocytes. (c) Flow cytometric analysis of thymic CD1d-αGalCer tetramer-positive NKT cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (d) Total cell numbers and percentages of CD1d-αGalCer tetramer-positive NKT cells in the spleen. For b and d, 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,b) or representative of two independent experiments with four to six mice per group (c,d).
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f2: Thymic T-cell differentiation in LUBAC-deficient mice.(a) Surface staining of CD4 and CD8 (upper panels), and FOXP3 and CD25 (lower panels) on thymocytes from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (b) Quantification of total cell numbers and percentages of CD4+ CD8+ and FOXP3+ thymocytes. (c) Flow cytometric analysis of thymic CD1d-αGalCer tetramer-positive NKT cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (d) Total cell numbers and percentages of CD1d-αGalCer tetramer-positive NKT cells in the spleen. For b and d, 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,b) or representative of two independent experiments with four to six mice per group (c,d).

Mentions: We tracked the origin of these T-cell defects to the thymus. The proportions of CD4+ and CD8+ SP thymocytes were significantly reduced in HoipΔCd4 and HoilΔCd4 mice but were found to be normal in Sharpincpdm mice (Fig. 2a,b and Supplementary Fig. 2). By contrast, the proportions and numbers of FOXP3+ Treg cells among CD4SP thymocytes were greatly diminished in all three strains, as were the CD25+ FOXP3− and CD25− FOXP3+ thymic Treg cell precursors (Fig. 2a,b). These data demonstrate that all three LUBAC components are required for the earliest checkpoint in Treg cell differentiation. NKT cells were almost undetectable in the thymus of HoipΔCd4 and HoilΔCd4 mice but were present in normal numbers in Sharpincpdm mice (Fig. 2c,d), demonstrating that this lineage has a dependency on LUBAC similar to that of conventional αβTCR T cells.


Linear ubiquitin chain assembly complex coordinates late thymic T-cell differentiation and regulatory T-cell homeostasis
Thymic T-cell differentiation in LUBAC-deficient mice.(a) Surface staining of CD4 and CD8 (upper panels), and FOXP3 and CD25 (lower panels) on thymocytes from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (b) Quantification of total cell numbers and percentages of CD4+ CD8+ and FOXP3+ thymocytes. (c) Flow cytometric analysis of thymic CD1d-αGalCer tetramer-positive NKT cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (d) Total cell numbers and percentages of CD1d-αGalCer tetramer-positive NKT cells in the spleen. For b and d, 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,b) or representative of two independent experiments with four to six mice per group (c,d).
© Copyright Policy - open-access
Related In: Results  -  Collection

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f2: Thymic T-cell differentiation in LUBAC-deficient mice.(a) Surface staining of CD4 and CD8 (upper panels), and FOXP3 and CD25 (lower panels) on thymocytes from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (b) Quantification of total cell numbers and percentages of CD4+ CD8+ and FOXP3+ thymocytes. (c) Flow cytometric analysis of thymic CD1d-αGalCer tetramer-positive NKT cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (d) Total cell numbers and percentages of CD1d-αGalCer tetramer-positive NKT cells in the spleen. For b and d, 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,b) or representative of two independent experiments with four to six mice per group (c,d).
Mentions: We tracked the origin of these T-cell defects to the thymus. The proportions of CD4+ and CD8+ SP thymocytes were significantly reduced in HoipΔCd4 and HoilΔCd4 mice but were found to be normal in Sharpincpdm mice (Fig. 2a,b and Supplementary Fig. 2). By contrast, the proportions and numbers of FOXP3+ Treg cells among CD4SP thymocytes were greatly diminished in all three strains, as were the CD25+ FOXP3− and CD25− FOXP3+ thymic Treg cell precursors (Fig. 2a,b). These data demonstrate that all three LUBAC components are required for the earliest checkpoint in Treg cell differentiation. NKT cells were almost undetectable in the thymus of HoipΔCd4 and HoilΔCd4 mice but were present in normal numbers in Sharpincpdm mice (Fig. 2c,d), demonstrating that this lineage has a dependency on LUBAC similar to that of conventional αβTCR T cells.

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.