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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

Ndfip1/Ndfip2-deficient CD4+ T cells cause increased colitis.(a–e) 0.5 × 106 sorted naive CD4+ T cells from WT, Ndfip2−/−, cKO and cDKO mice were transferred into 6-week-old Rag1−/− recipients. Mice were weighed twice weekly and killed 6 weeks after transfer when 20% weight loss was observed in multiple mice. Spleen weight and body weight were compared to generate an inflammation index (a) and colons were measured (b). H&E-stained sections of the distal colon were imaged on the × 20 objective, and crypt depth was quantified (c,d). Splenocytes were stained for intracellular IL-4 and IL-17 after ex vivo stimulation with PMA/ionomycin in the presence of BFA, and analysed by flow cytometry (e). Previously gated on live singlets, CD4+, dump gate-. Quantifications shown ±s.e.m. n=5–7 mice. Control (ctrl) mice did not receive T cells. P values calculated by ordinary 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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f4: Ndfip1/Ndfip2-deficient CD4+ T cells cause increased colitis.(a–e) 0.5 × 106 sorted naive CD4+ T cells from WT, Ndfip2−/−, cKO and cDKO mice were transferred into 6-week-old Rag1−/− recipients. Mice were weighed twice weekly and killed 6 weeks after transfer when 20% weight loss was observed in multiple mice. Spleen weight and body weight were compared to generate an inflammation index (a) and colons were measured (b). H&E-stained sections of the distal colon were imaged on the × 20 objective, and crypt depth was quantified (c,d). Splenocytes were stained for intracellular IL-4 and IL-17 after ex vivo stimulation with PMA/ionomycin in the presence of BFA, and analysed by flow cytometry (e). Previously gated on live singlets, CD4+, dump gate-. Quantifications shown ±s.e.m. n=5–7 mice. Control (ctrl) mice did not receive T cells. P values calculated by ordinary one-way ANOVA with Holm–Sidak test for multiple comparisons: *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Mentions: Our data indicate that both Ndfip1 and Ndfip2 control effector cell numbers and pathogenicity. To test this we transferred naive WT, Ndfip2−/−, Ndfip1 cKO and cDKO CD4+ T cells into Rag1−/− recipients to induce colitis. cDKO CD4+ T cells caused severe pathology: recipients had high spleen/body weight ratios, contracted colons and increased colon crypt depth (Fig. 4a–d). Recipients of Ndfip2−/− CD4+ T cells showed equivalent pathology to Ndfip1 cKO recipients—both cohorts developed more severe inflammation than WT cell recipients as measured by inflammation index, although colon histopathology, as quantified by crypt depth, was similar. Ndfip2−/− T cells, like WT cells, were IL-17A producers, while cDKO cells, like Ndfip1-deficient T cells, produced IL-4 (Fig. 4e). Thus, while Ndfip1 plays a dominant role in limiting TH2 differentiation, after initial activation both Ndfip1 and Ndfip2 limit the pathogenic potential of activated CD4+ T cells by limiting their expansion and function.


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)

Ndfip1/Ndfip2-deficient CD4+ T cells cause increased colitis.(a–e) 0.5 × 106 sorted naive CD4+ T cells from WT, Ndfip2−/−, cKO and cDKO mice were transferred into 6-week-old Rag1−/− recipients. Mice were weighed twice weekly and killed 6 weeks after transfer when 20% weight loss was observed in multiple mice. Spleen weight and body weight were compared to generate an inflammation index (a) and colons were measured (b). H&E-stained sections of the distal colon were imaged on the × 20 objective, and crypt depth was quantified (c,d). Splenocytes were stained for intracellular IL-4 and IL-17 after ex vivo stimulation with PMA/ionomycin in the presence of BFA, and analysed by flow cytometry (e). Previously gated on live singlets, CD4+, dump gate-. Quantifications shown ±s.e.m. n=5–7 mice. Control (ctrl) mice did not receive T cells. P values calculated by ordinary one-way ANOVA with Holm–Sidak test for multiple comparisons: *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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f4: Ndfip1/Ndfip2-deficient CD4+ T cells cause increased colitis.(a–e) 0.5 × 106 sorted naive CD4+ T cells from WT, Ndfip2−/−, cKO and cDKO mice were transferred into 6-week-old Rag1−/− recipients. Mice were weighed twice weekly and killed 6 weeks after transfer when 20% weight loss was observed in multiple mice. Spleen weight and body weight were compared to generate an inflammation index (a) and colons were measured (b). H&E-stained sections of the distal colon were imaged on the × 20 objective, and crypt depth was quantified (c,d). Splenocytes were stained for intracellular IL-4 and IL-17 after ex vivo stimulation with PMA/ionomycin in the presence of BFA, and analysed by flow cytometry (e). Previously gated on live singlets, CD4+, dump gate-. Quantifications shown ±s.e.m. n=5–7 mice. Control (ctrl) mice did not receive T cells. P values calculated by ordinary one-way ANOVA with Holm–Sidak test for multiple comparisons: *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.
Mentions: Our data indicate that both Ndfip1 and Ndfip2 control effector cell numbers and pathogenicity. To test this we transferred naive WT, Ndfip2−/−, Ndfip1 cKO and cDKO CD4+ T cells into Rag1−/− recipients to induce colitis. cDKO CD4+ T cells caused severe pathology: recipients had high spleen/body weight ratios, contracted colons and increased colon crypt depth (Fig. 4a–d). Recipients of Ndfip2−/− CD4+ T cells showed equivalent pathology to Ndfip1 cKO recipients—both cohorts developed more severe inflammation than WT cell recipients as measured by inflammation index, although colon histopathology, as quantified by crypt depth, was similar. Ndfip2−/− T cells, like WT cells, were IL-17A producers, while cDKO cells, like Ndfip1-deficient T cells, produced IL-4 (Fig. 4e). Thus, while Ndfip1 plays a dominant role in limiting TH2 differentiation, after initial activation both Ndfip1 and Ndfip2 limit the pathogenic potential of activated CD4+ T cells by limiting their expansion and function.

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