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Targeted inactivation of the IL-4 receptor alpha chain I4R motif promotes allergic airway inflammation.

Blaeser F, Bryce PJ, Ho N, Raman V, Dedeoglu F, Donaldson DD, Geha RS, Oettgen HC, Chatila TA - J. Exp. Med. (2003)

Bottom Line: The insulin/interleukin-4 (IL-4) receptor (I4R) motif mediates the association of insulin receptor substrate (IRS)-2 with the interleukin-4 (IL-4)Ralpha chain and transduces mitogenic signals in response to IL-4.The Y500F mutation abrogated IRS-2 phosphorylation and impaired IL-4-induced CD4+ T lymphocyte proliferation but left unperturbed Stat6 activation, up-regulation of IL-4-responsive gene products, and Th cell differentiation under Th2 polarizing conditions.These results define an important role for the I4R motif in regulating allergic inflammation.

View Article: PubMed Central - PubMed

Affiliation: Deparment of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110, USA.

ABSTRACT
The insulin/interleukin-4 (IL-4) receptor (I4R) motif mediates the association of insulin receptor substrate (IRS)-2 with the interleukin-4 (IL-4)Ralpha chain and transduces mitogenic signals in response to IL-4. Its physiological functions were analyzed in mice with a germline point mutation that changed the motif's effector tyrosine residue into phenylalanine (Y500F). The Y500F mutation abrogated IRS-2 phosphorylation and impaired IL-4-induced CD4+ T lymphocyte proliferation but left unperturbed Stat6 activation, up-regulation of IL-4-responsive gene products, and Th cell differentiation under Th2 polarizing conditions. However, in vivo the Y500F mutation was associated with increased allergen-induced IgE production, airway responsiveness, tissue eosinophilia, and mucus production. These results define an important role for the I4R motif in regulating allergic inflammation.

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Effect of the Y500F mutation on IL-4–induced lymphocyte proliferation and gene expression. (A) T cell proliferation. (Left) CD4+ splenic T cells of WT and homozygous Y500F littermate mice were cultured with plate-bound anti-TCRβ mAb (100 ng/ml) and IL-4 as indicated. Proliferation was assessed by 3H-thymidine incorporation. Results are mean responses of triplicate cultures ± SE and are representative of three experiments. (Right) Purified CD4+ T cells were cultured with PMA (25 ng/ml) and ionomycin (1 μM) or with plate-bound anti-TCRβ and IL-2 (100 U/ml). (B) B cell proliferation. (Left) Splenic B cells were cultured with anti-CD40 mAb (1 μg/ml) and IL-4 as indicated. (Right) Splenic B cells were cultured with IL-4 (10 ng/ml), anti-IgM antibodies (10 μg/ml), or IL-4 and anti-IgM antibodies as indicated. (C) IL-4–induced up-regulation of CD23 and MHC class II antigens. Splenic B cells were cultured for 48 h in the absence (thin line) or in the presence (thick line) of IL-4 at 50 ng/ml and then examined for CD23 and I-Ad expression by flow cytometry. Results are representative of five experiments.
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fig3: Effect of the Y500F mutation on IL-4–induced lymphocyte proliferation and gene expression. (A) T cell proliferation. (Left) CD4+ splenic T cells of WT and homozygous Y500F littermate mice were cultured with plate-bound anti-TCRβ mAb (100 ng/ml) and IL-4 as indicated. Proliferation was assessed by 3H-thymidine incorporation. Results are mean responses of triplicate cultures ± SE and are representative of three experiments. (Right) Purified CD4+ T cells were cultured with PMA (25 ng/ml) and ionomycin (1 μM) or with plate-bound anti-TCRβ and IL-2 (100 U/ml). (B) B cell proliferation. (Left) Splenic B cells were cultured with anti-CD40 mAb (1 μg/ml) and IL-4 as indicated. (Right) Splenic B cells were cultured with IL-4 (10 ng/ml), anti-IgM antibodies (10 μg/ml), or IL-4 and anti-IgM antibodies as indicated. (C) IL-4–induced up-regulation of CD23 and MHC class II antigens. Splenic B cells were cultured for 48 h in the absence (thin line) or in the presence (thick line) of IL-4 at 50 ng/ml and then examined for CD23 and I-Ad expression by flow cytometry. Results are representative of five experiments.

Mentions: The role of the Y500 residue in transducing mitogenic signals in lymphocytes was examined in splenic CD4+ T cells and B cells of WT and Y500F mutant mice. Fig. 3 A shows that in contrast to WT CD4+ T cells, which exhibited a modest proliferative response to IL-4, the response of mutant T cells was substantially decreased. Significantly, the Y500F mutation abrogated the comitogenic function of IL-4 in supporting T cell proliferation induced by anti-TCR antibodies. This effect was specific in that the response to stimulation with anti-TCR antibodies and IL-2 or with phorbol ester PMA and the calcium ionophore ionomycin was not affected.


Targeted inactivation of the IL-4 receptor alpha chain I4R motif promotes allergic airway inflammation.

Blaeser F, Bryce PJ, Ho N, Raman V, Dedeoglu F, Donaldson DD, Geha RS, Oettgen HC, Chatila TA - J. Exp. Med. (2003)

Effect of the Y500F mutation on IL-4–induced lymphocyte proliferation and gene expression. (A) T cell proliferation. (Left) CD4+ splenic T cells of WT and homozygous Y500F littermate mice were cultured with plate-bound anti-TCRβ mAb (100 ng/ml) and IL-4 as indicated. Proliferation was assessed by 3H-thymidine incorporation. Results are mean responses of triplicate cultures ± SE and are representative of three experiments. (Right) Purified CD4+ T cells were cultured with PMA (25 ng/ml) and ionomycin (1 μM) or with plate-bound anti-TCRβ and IL-2 (100 U/ml). (B) B cell proliferation. (Left) Splenic B cells were cultured with anti-CD40 mAb (1 μg/ml) and IL-4 as indicated. (Right) Splenic B cells were cultured with IL-4 (10 ng/ml), anti-IgM antibodies (10 μg/ml), or IL-4 and anti-IgM antibodies as indicated. (C) IL-4–induced up-regulation of CD23 and MHC class II antigens. Splenic B cells were cultured for 48 h in the absence (thin line) or in the presence (thick line) of IL-4 at 50 ng/ml and then examined for CD23 and I-Ad expression by flow cytometry. Results are representative of five experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2194235&req=5

fig3: Effect of the Y500F mutation on IL-4–induced lymphocyte proliferation and gene expression. (A) T cell proliferation. (Left) CD4+ splenic T cells of WT and homozygous Y500F littermate mice were cultured with plate-bound anti-TCRβ mAb (100 ng/ml) and IL-4 as indicated. Proliferation was assessed by 3H-thymidine incorporation. Results are mean responses of triplicate cultures ± SE and are representative of three experiments. (Right) Purified CD4+ T cells were cultured with PMA (25 ng/ml) and ionomycin (1 μM) or with plate-bound anti-TCRβ and IL-2 (100 U/ml). (B) B cell proliferation. (Left) Splenic B cells were cultured with anti-CD40 mAb (1 μg/ml) and IL-4 as indicated. (Right) Splenic B cells were cultured with IL-4 (10 ng/ml), anti-IgM antibodies (10 μg/ml), or IL-4 and anti-IgM antibodies as indicated. (C) IL-4–induced up-regulation of CD23 and MHC class II antigens. Splenic B cells were cultured for 48 h in the absence (thin line) or in the presence (thick line) of IL-4 at 50 ng/ml and then examined for CD23 and I-Ad expression by flow cytometry. Results are representative of five experiments.
Mentions: The role of the Y500 residue in transducing mitogenic signals in lymphocytes was examined in splenic CD4+ T cells and B cells of WT and Y500F mutant mice. Fig. 3 A shows that in contrast to WT CD4+ T cells, which exhibited a modest proliferative response to IL-4, the response of mutant T cells was substantially decreased. Significantly, the Y500F mutation abrogated the comitogenic function of IL-4 in supporting T cell proliferation induced by anti-TCR antibodies. This effect was specific in that the response to stimulation with anti-TCR antibodies and IL-2 or with phorbol ester PMA and the calcium ionophore ionomycin was not affected.

Bottom Line: The insulin/interleukin-4 (IL-4) receptor (I4R) motif mediates the association of insulin receptor substrate (IRS)-2 with the interleukin-4 (IL-4)Ralpha chain and transduces mitogenic signals in response to IL-4.The Y500F mutation abrogated IRS-2 phosphorylation and impaired IL-4-induced CD4+ T lymphocyte proliferation but left unperturbed Stat6 activation, up-regulation of IL-4-responsive gene products, and Th cell differentiation under Th2 polarizing conditions.These results define an important role for the I4R motif in regulating allergic inflammation.

View Article: PubMed Central - PubMed

Affiliation: Deparment of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110, USA.

ABSTRACT
The insulin/interleukin-4 (IL-4) receptor (I4R) motif mediates the association of insulin receptor substrate (IRS)-2 with the interleukin-4 (IL-4)Ralpha chain and transduces mitogenic signals in response to IL-4. Its physiological functions were analyzed in mice with a germline point mutation that changed the motif's effector tyrosine residue into phenylalanine (Y500F). The Y500F mutation abrogated IRS-2 phosphorylation and impaired IL-4-induced CD4+ T lymphocyte proliferation but left unperturbed Stat6 activation, up-regulation of IL-4-responsive gene products, and Th cell differentiation under Th2 polarizing conditions. However, in vivo the Y500F mutation was associated with increased allergen-induced IgE production, airway responsiveness, tissue eosinophilia, and mucus production. These results define an important role for the I4R motif in regulating allergic inflammation.

Show MeSH
Related in: MedlinePlus