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Role of Tyk-2 in Th9 and Th17 cells in allergic asthma.

Übel C, Graser A, Koch S, Rieker RJ, Lehr HA, Müller M, Finotto S - Sci Rep (2014)

Bottom Line: In a murine model of allergic asthma, we found that Tyk-2((-/-)) asthmatic mice have induced peribronchial collagen deposition, mucosal type mast cells in the lung, IRF4 and hyperproliferative lung Th2 CD4(+) effector T cells over-expressing IL-3, IL-4, IL-5, IL-10 and IL-13.We also observed increased Th9 cells expressing IL-9 and IL-10 as well as T helper cells expressing IL-6, IL-10 and IL-21 with a defect in IL-17A and IL-17F production.Finally, in vivo treatment with rIL-17A inhibited local CD4(+)CD25(+)Foxp3(+) T regulatory cells as well as Th2 cytokines without affecting IL-9 in the lung.

View Article: PubMed Central - PubMed

Affiliation: 1] Laboratory of Cellular and Molecular Immunology of the Lung, Institute of Molecular Pneumology, University of Erlangen-Nürnberg, 91054 Erlangen, Germany [2].

ABSTRACT
In a murine model of allergic asthma, we found that Tyk-2((-/-)) asthmatic mice have induced peribronchial collagen deposition, mucosal type mast cells in the lung, IRF4 and hyperproliferative lung Th2 CD4(+) effector T cells over-expressing IL-3, IL-4, IL-5, IL-10 and IL-13. We also observed increased Th9 cells expressing IL-9 and IL-10 as well as T helper cells expressing IL-6, IL-10 and IL-21 with a defect in IL-17A and IL-17F production. This T helper phenotype was accompanied by increased SOCS3 in the lung of Tyk-2 deficient asthmatic mice. Finally, in vivo treatment with rIL-17A inhibited local CD4(+)CD25(+)Foxp3(+) T regulatory cells as well as Th2 cytokines without affecting IL-9 in the lung. These results suggest a role of Tyk-2 in different subsets of T helper cells mediated by SOCS3 regulation that is relevant for the treatment of asthma, cancer and autoimmune diseases.

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Treatment with rIL-17A led to down-regulation of T regulatory cells in Tyk-2 deficient mice.(a). Mice were treated with OVA alone and OVA + rIL-17A as shown in the experimental design. (b).–(d). Lung CD4+T cells from OVA alone and OVA + rIL-17A treated mice were isolated and incubated for 24 h with α-CD3 and α-CD28. The supernatants were analyzed by ELISA for IL-4, IL-5 and IL-13 ((b):IL-4: n = 3–4; p = 0.004; p = 0.003; (c): IL-5: n = 3–4; p = 0.0002; p = 0.040; (d): IL-13: n = 3–4; p = 0.002; p = 0.018; p = 0.017). (e). BALF was retrieved and BAL cells were incubated with antibodies against CD3, CD45R, CCR3 and Gr-1 and subsequently analyzed by flow cytometry. CD3−CD45R−CCR3−Gr-1+ cells were counted as neutrophils ((b). n = 3–4; p = 0.039; p = 0.029; p = 0.036; p = 0.015). (f). Total lung cells from OVA and OVA + rIL-17A treated mice were incubated with antibodies against CD4, CD25 and Foxp3 and analyzed by flow cytometry (n = 3–4; p = 0.001; p = 0.002). (g). Lung CD4+T cells from OVA alone and OVA + rIL-17A treated mice were isolated and incubated for 24 h with α-CD3 and α-CD28. The supernatants were analyzed by ELISA for IL-9 (n = 2–3; p = 0.0004; p = 0.016).
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f5: Treatment with rIL-17A led to down-regulation of T regulatory cells in Tyk-2 deficient mice.(a). Mice were treated with OVA alone and OVA + rIL-17A as shown in the experimental design. (b).–(d). Lung CD4+T cells from OVA alone and OVA + rIL-17A treated mice were isolated and incubated for 24 h with α-CD3 and α-CD28. The supernatants were analyzed by ELISA for IL-4, IL-5 and IL-13 ((b):IL-4: n = 3–4; p = 0.004; p = 0.003; (c): IL-5: n = 3–4; p = 0.0002; p = 0.040; (d): IL-13: n = 3–4; p = 0.002; p = 0.018; p = 0.017). (e). BALF was retrieved and BAL cells were incubated with antibodies against CD3, CD45R, CCR3 and Gr-1 and subsequently analyzed by flow cytometry. CD3−CD45R−CCR3−Gr-1+ cells were counted as neutrophils ((b). n = 3–4; p = 0.039; p = 0.029; p = 0.036; p = 0.015). (f). Total lung cells from OVA and OVA + rIL-17A treated mice were incubated with antibodies against CD4, CD25 and Foxp3 and analyzed by flow cytometry (n = 3–4; p = 0.001; p = 0.002). (g). Lung CD4+T cells from OVA alone and OVA + rIL-17A treated mice were isolated and incubated for 24 h with α-CD3 and α-CD28. The supernatants were analyzed by ELISA for IL-9 (n = 2–3; p = 0.0004; p = 0.016).

Mentions: Intranasal application of rIL-17A (Fig. 5a) led to a down regulation of the Th2 cytokines IL-4, IL-5 and IL-13 only in Tyk-2(−/−) mice (Fig. 5b–d, respectively). IL-17A is important for the recruitment of neutrophils to the lung during allergic airway disease33. We thus analyzed the number of neutrophils in BAL cells of naïve mice and after allergen challenge. We detected a defect of neutrophils in the lung of Tyk-2(−/−) mice after allergen challenge. In vivo intranasal application of recombinant IL-17A induced neutrophils in wild type and less inTyk-2(−/−) mice (Fig. 5e). The latter result is consistent with a role of IL-17A on neutrophilia and with defective IL-17A in the lung of Tyk-2 deficient mice. Application of IL-17A and OVA led to a down-regulation of T regulatory cells in Tyk-2(−/−) mice, whereas the number of these cells remained unchanged in wild-type mice (Fig. 5f). Thus, Tyk-2(−/−) mice had fewer regulatory T cells after IL-17A treatment than wild type mice consistent with a role of Tyk-2 in the generation of T regulatory cells downstream of the IL-17A receptor.


Role of Tyk-2 in Th9 and Th17 cells in allergic asthma.

Übel C, Graser A, Koch S, Rieker RJ, Lehr HA, Müller M, Finotto S - Sci Rep (2014)

Treatment with rIL-17A led to down-regulation of T regulatory cells in Tyk-2 deficient mice.(a). Mice were treated with OVA alone and OVA + rIL-17A as shown in the experimental design. (b).–(d). Lung CD4+T cells from OVA alone and OVA + rIL-17A treated mice were isolated and incubated for 24 h with α-CD3 and α-CD28. The supernatants were analyzed by ELISA for IL-4, IL-5 and IL-13 ((b):IL-4: n = 3–4; p = 0.004; p = 0.003; (c): IL-5: n = 3–4; p = 0.0002; p = 0.040; (d): IL-13: n = 3–4; p = 0.002; p = 0.018; p = 0.017). (e). BALF was retrieved and BAL cells were incubated with antibodies against CD3, CD45R, CCR3 and Gr-1 and subsequently analyzed by flow cytometry. CD3−CD45R−CCR3−Gr-1+ cells were counted as neutrophils ((b). n = 3–4; p = 0.039; p = 0.029; p = 0.036; p = 0.015). (f). Total lung cells from OVA and OVA + rIL-17A treated mice were incubated with antibodies against CD4, CD25 and Foxp3 and analyzed by flow cytometry (n = 3–4; p = 0.001; p = 0.002). (g). Lung CD4+T cells from OVA alone and OVA + rIL-17A treated mice were isolated and incubated for 24 h with α-CD3 and α-CD28. The supernatants were analyzed by ELISA for IL-9 (n = 2–3; p = 0.0004; p = 0.016).
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f5: Treatment with rIL-17A led to down-regulation of T regulatory cells in Tyk-2 deficient mice.(a). Mice were treated with OVA alone and OVA + rIL-17A as shown in the experimental design. (b).–(d). Lung CD4+T cells from OVA alone and OVA + rIL-17A treated mice were isolated and incubated for 24 h with α-CD3 and α-CD28. The supernatants were analyzed by ELISA for IL-4, IL-5 and IL-13 ((b):IL-4: n = 3–4; p = 0.004; p = 0.003; (c): IL-5: n = 3–4; p = 0.0002; p = 0.040; (d): IL-13: n = 3–4; p = 0.002; p = 0.018; p = 0.017). (e). BALF was retrieved and BAL cells were incubated with antibodies against CD3, CD45R, CCR3 and Gr-1 and subsequently analyzed by flow cytometry. CD3−CD45R−CCR3−Gr-1+ cells were counted as neutrophils ((b). n = 3–4; p = 0.039; p = 0.029; p = 0.036; p = 0.015). (f). Total lung cells from OVA and OVA + rIL-17A treated mice were incubated with antibodies against CD4, CD25 and Foxp3 and analyzed by flow cytometry (n = 3–4; p = 0.001; p = 0.002). (g). Lung CD4+T cells from OVA alone and OVA + rIL-17A treated mice were isolated and incubated for 24 h with α-CD3 and α-CD28. The supernatants were analyzed by ELISA for IL-9 (n = 2–3; p = 0.0004; p = 0.016).
Mentions: Intranasal application of rIL-17A (Fig. 5a) led to a down regulation of the Th2 cytokines IL-4, IL-5 and IL-13 only in Tyk-2(−/−) mice (Fig. 5b–d, respectively). IL-17A is important for the recruitment of neutrophils to the lung during allergic airway disease33. We thus analyzed the number of neutrophils in BAL cells of naïve mice and after allergen challenge. We detected a defect of neutrophils in the lung of Tyk-2(−/−) mice after allergen challenge. In vivo intranasal application of recombinant IL-17A induced neutrophils in wild type and less inTyk-2(−/−) mice (Fig. 5e). The latter result is consistent with a role of IL-17A on neutrophilia and with defective IL-17A in the lung of Tyk-2 deficient mice. Application of IL-17A and OVA led to a down-regulation of T regulatory cells in Tyk-2(−/−) mice, whereas the number of these cells remained unchanged in wild-type mice (Fig. 5f). Thus, Tyk-2(−/−) mice had fewer regulatory T cells after IL-17A treatment than wild type mice consistent with a role of Tyk-2 in the generation of T regulatory cells downstream of the IL-17A receptor.

Bottom Line: In a murine model of allergic asthma, we found that Tyk-2((-/-)) asthmatic mice have induced peribronchial collagen deposition, mucosal type mast cells in the lung, IRF4 and hyperproliferative lung Th2 CD4(+) effector T cells over-expressing IL-3, IL-4, IL-5, IL-10 and IL-13.We also observed increased Th9 cells expressing IL-9 and IL-10 as well as T helper cells expressing IL-6, IL-10 and IL-21 with a defect in IL-17A and IL-17F production.Finally, in vivo treatment with rIL-17A inhibited local CD4(+)CD25(+)Foxp3(+) T regulatory cells as well as Th2 cytokines without affecting IL-9 in the lung.

View Article: PubMed Central - PubMed

Affiliation: 1] Laboratory of Cellular and Molecular Immunology of the Lung, Institute of Molecular Pneumology, University of Erlangen-Nürnberg, 91054 Erlangen, Germany [2].

ABSTRACT
In a murine model of allergic asthma, we found that Tyk-2((-/-)) asthmatic mice have induced peribronchial collagen deposition, mucosal type mast cells in the lung, IRF4 and hyperproliferative lung Th2 CD4(+) effector T cells over-expressing IL-3, IL-4, IL-5, IL-10 and IL-13. We also observed increased Th9 cells expressing IL-9 and IL-10 as well as T helper cells expressing IL-6, IL-10 and IL-21 with a defect in IL-17A and IL-17F production. This T helper phenotype was accompanied by increased SOCS3 in the lung of Tyk-2 deficient asthmatic mice. Finally, in vivo treatment with rIL-17A inhibited local CD4(+)CD25(+)Foxp3(+) T regulatory cells as well as Th2 cytokines without affecting IL-9 in the lung. These results suggest a role of Tyk-2 in different subsets of T helper cells mediated by SOCS3 regulation that is relevant for the treatment of asthma, cancer and autoimmune diseases.

Show MeSH
Related in: MedlinePlus