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The Runx1 transcription factor inhibits the differentiation of naive CD4+ T cells into the Th2 lineage by repressing GATA3 expression.

Komine O, Hayashi K, Natsume W, Watanabe T, Seki Y, Seki N, Yagi R, Sukzuki W, Tamauchi H, Hozumi K, Habu S, Kubo M, Satake M - J. Exp. Med. (2003)

Bottom Line: In contrast, transduction of Runx1 into wild-type T cells caused a complete attenuation of Th2 differentiation and was accompanied by the cessation of GATA3 expression.Furthermore, endogenous expression of Runx1 in naive T cells declined after T cell receptor stimulation, at the same time that expression of GATA3 increased.We conclude that Runx1 plays a novel role as a negative regulator of GATA3 expression, thereby inhibiting the Th2 cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Research Institute for Biological Sciences, Tokyo University of Science, Noda, Japan.

ABSTRACT
Differentiation of naive CD4+ T cells into helper T (Th) cells is controlled by a combination of several transcriptional factors. In this study, we examined the functional role of the Runx1 transcription factor in Th cell differentiation. Naive T cells from transgenic mice expressing a dominant interfering form of Runx1 exhibited enhanced interleukin 4 production and efficient Th2 differentiation. In contrast, transduction of Runx1 into wild-type T cells caused a complete attenuation of Th2 differentiation and was accompanied by the cessation of GATA3 expression. Furthermore, endogenous expression of Runx1 in naive T cells declined after T cell receptor stimulation, at the same time that expression of GATA3 increased. We conclude that Runx1 plays a novel role as a negative regulator of GATA3 expression, thereby inhibiting the Th2 cell differentiation.

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Effect of IL-4 supplementation on the production of various cytokines from Runx1-transduced cells. (A) Naive CD4+ T cells were isolated from the spleens of BALB/c mice, stimulated with anti-CD3/anti-CD28 antibodies and infected by retroviruses carrying pMX or pMX-Runx1. After culture in the presence of IL-2 alone or of both IL-2 and IL-4, the cells were washed, reactivated via TCR, and processed for flow cytometrical analysis of intracellular IL-4, IFN-γ, and GFP. The numbers represent the percentages of cells in each quadrant. (B) The GFP+ population was sorted from the retrovirus-infected cells and reactivated via TCR. The amount of cytokines secreted into the culture supernatant was measured by ELISA. Data are representative of two independent experiments.
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fig3: Effect of IL-4 supplementation on the production of various cytokines from Runx1-transduced cells. (A) Naive CD4+ T cells were isolated from the spleens of BALB/c mice, stimulated with anti-CD3/anti-CD28 antibodies and infected by retroviruses carrying pMX or pMX-Runx1. After culture in the presence of IL-2 alone or of both IL-2 and IL-4, the cells were washed, reactivated via TCR, and processed for flow cytometrical analysis of intracellular IL-4, IFN-γ, and GFP. The numbers represent the percentages of cells in each quadrant. (B) The GFP+ population was sorted from the retrovirus-infected cells and reactivated via TCR. The amount of cytokines secreted into the culture supernatant was measured by ELISA. Data are representative of two independent experiments.

Mentions: We next examined whether overexpression of Runx1 is inhibitory for Th2-cell differentiation even in a Th2-favorable cytokine environment. CD4+ T cells were stimulated with anti-CD3/anti-CD28 antibodies, infected by retroviruses, and cultured in the presence or absence of an excessive amount of exogenously added IL-4. The cells were restimulated via TCR and analyzed for cytokine production using flow cytometry (Fig. 3 A). In the pMX-infected cells, addition of IL-4 increased the ratio of IL-4+ cells among the GFP+ population as expected. IL-4 production was much lower in the Runx1-overexpressing cells than in the pMX-infected cells, and IL-4 supplementation did not restore IL-4 production in these cells. The amount of cytokines secreted from the GFP+ population was also measured by ELISA (Fig. 3 B). Addition of IL-4 enhanced the secretion of both IL-4 and IL-5 from the pMX-infected cells. In the Runx1-overexpressing cells, however, production of IL-4 and IL-5 did not recover even after addition of IL-4. Thus, the overexpression of Runx1 can block TCR-stimulated CD4+ cells from differentiating into the Th2 lineage even under a Th2-favorable culture condition and this inhibitory effect of Runx1 on Th2 differentiation is not due to the paucity of IL-4 itself.


The Runx1 transcription factor inhibits the differentiation of naive CD4+ T cells into the Th2 lineage by repressing GATA3 expression.

Komine O, Hayashi K, Natsume W, Watanabe T, Seki Y, Seki N, Yagi R, Sukzuki W, Tamauchi H, Hozumi K, Habu S, Kubo M, Satake M - J. Exp. Med. (2003)

Effect of IL-4 supplementation on the production of various cytokines from Runx1-transduced cells. (A) Naive CD4+ T cells were isolated from the spleens of BALB/c mice, stimulated with anti-CD3/anti-CD28 antibodies and infected by retroviruses carrying pMX or pMX-Runx1. After culture in the presence of IL-2 alone or of both IL-2 and IL-4, the cells were washed, reactivated via TCR, and processed for flow cytometrical analysis of intracellular IL-4, IFN-γ, and GFP. The numbers represent the percentages of cells in each quadrant. (B) The GFP+ population was sorted from the retrovirus-infected cells and reactivated via TCR. The amount of cytokines secreted into the culture supernatant was measured by ELISA. Data are representative of two independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196077&req=5

fig3: Effect of IL-4 supplementation on the production of various cytokines from Runx1-transduced cells. (A) Naive CD4+ T cells were isolated from the spleens of BALB/c mice, stimulated with anti-CD3/anti-CD28 antibodies and infected by retroviruses carrying pMX or pMX-Runx1. After culture in the presence of IL-2 alone or of both IL-2 and IL-4, the cells were washed, reactivated via TCR, and processed for flow cytometrical analysis of intracellular IL-4, IFN-γ, and GFP. The numbers represent the percentages of cells in each quadrant. (B) The GFP+ population was sorted from the retrovirus-infected cells and reactivated via TCR. The amount of cytokines secreted into the culture supernatant was measured by ELISA. Data are representative of two independent experiments.
Mentions: We next examined whether overexpression of Runx1 is inhibitory for Th2-cell differentiation even in a Th2-favorable cytokine environment. CD4+ T cells were stimulated with anti-CD3/anti-CD28 antibodies, infected by retroviruses, and cultured in the presence or absence of an excessive amount of exogenously added IL-4. The cells were restimulated via TCR and analyzed for cytokine production using flow cytometry (Fig. 3 A). In the pMX-infected cells, addition of IL-4 increased the ratio of IL-4+ cells among the GFP+ population as expected. IL-4 production was much lower in the Runx1-overexpressing cells than in the pMX-infected cells, and IL-4 supplementation did not restore IL-4 production in these cells. The amount of cytokines secreted from the GFP+ population was also measured by ELISA (Fig. 3 B). Addition of IL-4 enhanced the secretion of both IL-4 and IL-5 from the pMX-infected cells. In the Runx1-overexpressing cells, however, production of IL-4 and IL-5 did not recover even after addition of IL-4. Thus, the overexpression of Runx1 can block TCR-stimulated CD4+ cells from differentiating into the Th2 lineage even under a Th2-favorable culture condition and this inhibitory effect of Runx1 on Th2 differentiation is not due to the paucity of IL-4 itself.

Bottom Line: In contrast, transduction of Runx1 into wild-type T cells caused a complete attenuation of Th2 differentiation and was accompanied by the cessation of GATA3 expression.Furthermore, endogenous expression of Runx1 in naive T cells declined after T cell receptor stimulation, at the same time that expression of GATA3 increased.We conclude that Runx1 plays a novel role as a negative regulator of GATA3 expression, thereby inhibiting the Th2 cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Research Institute for Biological Sciences, Tokyo University of Science, Noda, Japan.

ABSTRACT
Differentiation of naive CD4+ T cells into helper T (Th) cells is controlled by a combination of several transcriptional factors. In this study, we examined the functional role of the Runx1 transcription factor in Th cell differentiation. Naive T cells from transgenic mice expressing a dominant interfering form of Runx1 exhibited enhanced interleukin 4 production and efficient Th2 differentiation. In contrast, transduction of Runx1 into wild-type T cells caused a complete attenuation of Th2 differentiation and was accompanied by the cessation of GATA3 expression. Furthermore, endogenous expression of Runx1 in naive T cells declined after T cell receptor stimulation, at the same time that expression of GATA3 increased. We conclude that Runx1 plays a novel role as a negative regulator of GATA3 expression, thereby inhibiting the Th2 cell differentiation.

Show MeSH