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Ndfip-mediated degradation of Jak1 tunes cytokine signalling to limit expansion of CD4+ effector T cells.

O'Leary CE, Riling CR, Spruce LA, Ding H, Kumar S, Deng G, Liu Y, Seeholzer SH, Oliver PM - Nat Commun (2016)

Bottom Line: Nedd4 family E3 ubiquitin ligases have been shown to restrict T-cell function and impact T-cell differentiation.We show here that Ndfip1 and Ndfip2, activators of Nedd4 family ligases, together limit accumulation and function of effector CD4+ T cells.In wild-type cells, Jak1 degradation lessens CD4+ cell sensitivity to cytokines during TCR stimulation, while in Ndfip-deficient cells cytokine responsiveness persists, promoting increased expansion and survival of pathogenic effector T cells.

View Article: PubMed Central - PubMed

Affiliation: Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.

ABSTRACT
Nedd4 family E3 ubiquitin ligases have been shown to restrict T-cell function and impact T-cell differentiation. We show here that Ndfip1 and Ndfip2, activators of Nedd4 family ligases, together limit accumulation and function of effector CD4+ T cells. Using a three-part proteomics approach in primary T cells, we identify stabilization of Jak1 in Ndfip1/2-deficient T cells stimulated through the TCR. Jak1 degradation is aborted in activated T cells that lack Ndfips. In wild-type cells, Jak1 degradation lessens CD4+ cell sensitivity to cytokines during TCR stimulation, while in Ndfip-deficient cells cytokine responsiveness persists, promoting increased expansion and survival of pathogenic effector T cells. Thus, Ndfip1/Ndfip2 regulate the cross talk between the T-cell receptor and cytokine signalling pathways to limit inappropriate T-cell responses.

No MeSH data available.


Related in: MedlinePlus

Ndfip2 deficiency exacerbates inflammation in Ndfip1fl/flCD4Cre+ mice.(a) H&E-stained sections of oesophagus, lung and skin from representative 8-week-old control, Ndfip1fl/flCD4 Cre+ (cKO) and Ndfip2−/− Ndfip1fl/flCD4 Cre+ (cDKO) mice (bar represents 100 μm). (b,c) Body weight (b) and (c) spleen count of 5–7-week-old WT, cKO, Ndfip2−/− and cDKO mice. Mean±s.e.m. n=5–15 mice. (d,e) Representative flow cytometry analysis of splenic CD3+CD4+ T cells, showing (d) expression CD44 and CD62L and (e) intracellular levels of IL-4 after ex vivo stimulation with PMA/ionomycin in the presence of BFA. (f–i) Quantification of (f) the number of CD4+ T cells, (g) naive CD62Lhigh CD4+ T cells, (h) CD44+ CD4+ T cells and (i) IL-4+ CD4s from spleen analysed by flow cytometry. Mean±s.e.m., n=7–12 mice. (j,k) Quantification of percent IL-4+ or IFNγ+ CD4+ T cells among CD44 high T cells. Mean±s.e.m., n=4–7 mice. P values calculated by one-way ANOVA with Holm–Sidak test for multiple comparisons: *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.
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f2: Ndfip2 deficiency exacerbates inflammation in Ndfip1fl/flCD4Cre+ mice.(a) H&E-stained sections of oesophagus, lung and skin from representative 8-week-old control, Ndfip1fl/flCD4 Cre+ (cKO) and Ndfip2−/− Ndfip1fl/flCD4 Cre+ (cDKO) mice (bar represents 100 μm). (b,c) Body weight (b) and (c) spleen count of 5–7-week-old WT, cKO, Ndfip2−/− and cDKO mice. Mean±s.e.m. n=5–15 mice. (d,e) Representative flow cytometry analysis of splenic CD3+CD4+ T cells, showing (d) expression CD44 and CD62L and (e) intracellular levels of IL-4 after ex vivo stimulation with PMA/ionomycin in the presence of BFA. (f–i) Quantification of (f) the number of CD4+ T cells, (g) naive CD62Lhigh CD4+ T cells, (h) CD44+ CD4+ T cells and (i) IL-4+ CD4s from spleen analysed by flow cytometry. Mean±s.e.m., n=7–12 mice. (j,k) Quantification of percent IL-4+ or IFNγ+ CD4+ T cells among CD44 high T cells. Mean±s.e.m., n=4–7 mice. P values calculated by one-way ANOVA with Holm–Sidak test for multiple comparisons: *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Mentions: We observed that Ndfip2−/−Ndfip1fl/flCD4 Cre+ (referred to as cDKO) mice showed increased inflammation at barrier surfaces, increased spleen size and decreased body weight relative to age-matched cKO, Ndfip2−/− and control mice (Fig. 2a–c). Splenic CD4+ T cells from cDKO mice were more likely to express CD44 (Fig. 2d) and produce IL-4 (Fig. 2e,i). We observed a significantly increased number of CD4+ T cells in spleens from cDKO mice (Fig. 2f) due largely to an increased number of CD44+ CD4+ T cells (Fig. 2g); the number of CD62L+ CD4+ T cells in cDKO spleens was not statistically different from cKO and control mice (Fig. 2h).


Ndfip-mediated degradation of Jak1 tunes cytokine signalling to limit expansion of CD4+ effector T cells.

O'Leary CE, Riling CR, Spruce LA, Ding H, Kumar S, Deng G, Liu Y, Seeholzer SH, Oliver PM - Nat Commun (2016)

Ndfip2 deficiency exacerbates inflammation in Ndfip1fl/flCD4Cre+ mice.(a) H&E-stained sections of oesophagus, lung and skin from representative 8-week-old control, Ndfip1fl/flCD4 Cre+ (cKO) and Ndfip2−/− Ndfip1fl/flCD4 Cre+ (cDKO) mice (bar represents 100 μm). (b,c) Body weight (b) and (c) spleen count of 5–7-week-old WT, cKO, Ndfip2−/− and cDKO mice. Mean±s.e.m. n=5–15 mice. (d,e) Representative flow cytometry analysis of splenic CD3+CD4+ T cells, showing (d) expression CD44 and CD62L and (e) intracellular levels of IL-4 after ex vivo stimulation with PMA/ionomycin in the presence of BFA. (f–i) Quantification of (f) the number of CD4+ T cells, (g) naive CD62Lhigh CD4+ T cells, (h) CD44+ CD4+ T cells and (i) IL-4+ CD4s from spleen analysed by flow cytometry. Mean±s.e.m., n=7–12 mice. (j,k) Quantification of percent IL-4+ or IFNγ+ CD4+ T cells among CD44 high T cells. Mean±s.e.m., n=4–7 mice. P values calculated by one-way ANOVA with Holm–Sidak test for multiple comparisons: *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.
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f2: Ndfip2 deficiency exacerbates inflammation in Ndfip1fl/flCD4Cre+ mice.(a) H&E-stained sections of oesophagus, lung and skin from representative 8-week-old control, Ndfip1fl/flCD4 Cre+ (cKO) and Ndfip2−/− Ndfip1fl/flCD4 Cre+ (cDKO) mice (bar represents 100 μm). (b,c) Body weight (b) and (c) spleen count of 5–7-week-old WT, cKO, Ndfip2−/− and cDKO mice. Mean±s.e.m. n=5–15 mice. (d,e) Representative flow cytometry analysis of splenic CD3+CD4+ T cells, showing (d) expression CD44 and CD62L and (e) intracellular levels of IL-4 after ex vivo stimulation with PMA/ionomycin in the presence of BFA. (f–i) Quantification of (f) the number of CD4+ T cells, (g) naive CD62Lhigh CD4+ T cells, (h) CD44+ CD4+ T cells and (i) IL-4+ CD4s from spleen analysed by flow cytometry. Mean±s.e.m., n=7–12 mice. (j,k) Quantification of percent IL-4+ or IFNγ+ CD4+ T cells among CD44 high T cells. Mean±s.e.m., n=4–7 mice. P values calculated by one-way ANOVA with Holm–Sidak test for multiple comparisons: *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.
Mentions: We observed that Ndfip2−/−Ndfip1fl/flCD4 Cre+ (referred to as cDKO) mice showed increased inflammation at barrier surfaces, increased spleen size and decreased body weight relative to age-matched cKO, Ndfip2−/− and control mice (Fig. 2a–c). Splenic CD4+ T cells from cDKO mice were more likely to express CD44 (Fig. 2d) and produce IL-4 (Fig. 2e,i). We observed a significantly increased number of CD4+ T cells in spleens from cDKO mice (Fig. 2f) due largely to an increased number of CD44+ CD4+ T cells (Fig. 2g); the number of CD62L+ CD4+ T cells in cDKO spleens was not statistically different from cKO and control mice (Fig. 2h).

Bottom Line: Nedd4 family E3 ubiquitin ligases have been shown to restrict T-cell function and impact T-cell differentiation.We show here that Ndfip1 and Ndfip2, activators of Nedd4 family ligases, together limit accumulation and function of effector CD4+ T cells.In wild-type cells, Jak1 degradation lessens CD4+ cell sensitivity to cytokines during TCR stimulation, while in Ndfip-deficient cells cytokine responsiveness persists, promoting increased expansion and survival of pathogenic effector T cells.

View Article: PubMed Central - PubMed

Affiliation: Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.

ABSTRACT
Nedd4 family E3 ubiquitin ligases have been shown to restrict T-cell function and impact T-cell differentiation. We show here that Ndfip1 and Ndfip2, activators of Nedd4 family ligases, together limit accumulation and function of effector CD4+ T cells. Using a three-part proteomics approach in primary T cells, we identify stabilization of Jak1 in Ndfip1/2-deficient T cells stimulated through the TCR. Jak1 degradation is aborted in activated T cells that lack Ndfips. In wild-type cells, Jak1 degradation lessens CD4+ cell sensitivity to cytokines during TCR stimulation, while in Ndfip-deficient cells cytokine responsiveness persists, promoting increased expansion and survival of pathogenic effector T cells. Thus, Ndfip1/Ndfip2 regulate the cross talk between the T-cell receptor and cytokine signalling pathways to limit inappropriate T-cell responses.

No MeSH data available.


Related in: MedlinePlus