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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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Production of Th2- and Th1-type cytokines from wild-type and Runt-transgenic CD4+ T cells. (A) Naive CD4+ T cells were isolated from the spleens of wild-type and Runt–transgenic mice and stimulated with anti-CD3/anti-CD28 antibodies, and the culture supernatants were collected at the indicated times after stimulation. (B) The TCR-stimulated cells were cultured in the presence of IL-2 for 5 d, washed with fresh media, and restimulated via TCR. The culture supernatants were collected after 24 h. (C) The cells were cultured for four days in the Th2- or Th1-inducing condition. The cells were washed with fresh media and restimulated via TCR. The culture supernatants were collected after 24 h. (D) Same as C, but note the difference of scale used in C and here. The amounts of cytokines secreted into the supernatants were measured by ELISA and their averages and standard deviations are shown. Data are representative of four independent experiments.
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fig1: Production of Th2- and Th1-type cytokines from wild-type and Runt-transgenic CD4+ T cells. (A) Naive CD4+ T cells were isolated from the spleens of wild-type and Runt–transgenic mice and stimulated with anti-CD3/anti-CD28 antibodies, and the culture supernatants were collected at the indicated times after stimulation. (B) The TCR-stimulated cells were cultured in the presence of IL-2 for 5 d, washed with fresh media, and restimulated via TCR. The culture supernatants were collected after 24 h. (C) The cells were cultured for four days in the Th2- or Th1-inducing condition. The cells were washed with fresh media and restimulated via TCR. The culture supernatants were collected after 24 h. (D) Same as C, but note the difference of scale used in C and here. The amounts of cytokines secreted into the supernatants were measured by ELISA and their averages and standard deviations are shown. Data are representative of four independent experiments.

Mentions: We previously established mouse lines that express a DNA binding domain of Runx1 from a transgene in a T-lineage-specific way (24). This Runt domain is known to function in a dominant interfering fashion against the Runx1 protein that is expressed endogenously in T lymphocytes. In the present study, we investigated the role of the Runx1 transcription factor in Th cell differentiation using these Runt-transgenic mice. Naive CD4+ T cells were isolated from the spleens of wild-type and Runt-transgenic mice and stimulated with anti-CD3/anti-CD28 antibodies. The culture supernatants were collected and the amount of cytokines secreted was measured by ELISA (Fig. 1 A). The Runt-transgenic cells produced five times as much IL-4 as the wild-type cells at 72 h after incubation. Similarly, the Runt-transgenic cells produced 1.7 and 3.4 times as much IL-5 and IL-10, respectively, as the wild-type cells. The Runt-transgenic cells thus exhibited enhanced production of Th2–type cytokines during an early phase of TCR activation. In contrast, secretion of IFN-γ and IL-2 from the Runt-transgenic cells was decreased slightly and markedly, respectively, compared with the wild-type cells. A similar result as that shown in Fig. 1 A was obtained when the TCR-stimulated cells were incubated in the presence of IL-2 for 5 d, washed with fresh media, and restimulated via TCR (Fig. 1 B). Under this neutral culture condition where neither IL-4 nor IL-12 was added to media, the Runt-transgenic cells secreted several fold more amount of Th2-type cytokines and less amount of Th1-type cytokines than the wild-type cells.


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)

Production of Th2- and Th1-type cytokines from wild-type and Runt-transgenic CD4+ T cells. (A) Naive CD4+ T cells were isolated from the spleens of wild-type and Runt–transgenic mice and stimulated with anti-CD3/anti-CD28 antibodies, and the culture supernatants were collected at the indicated times after stimulation. (B) The TCR-stimulated cells were cultured in the presence of IL-2 for 5 d, washed with fresh media, and restimulated via TCR. The culture supernatants were collected after 24 h. (C) The cells were cultured for four days in the Th2- or Th1-inducing condition. The cells were washed with fresh media and restimulated via TCR. The culture supernatants were collected after 24 h. (D) Same as C, but note the difference of scale used in C and here. The amounts of cytokines secreted into the supernatants were measured by ELISA and their averages and standard deviations are shown. Data are representative of four independent experiments.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2196077&req=5

fig1: Production of Th2- and Th1-type cytokines from wild-type and Runt-transgenic CD4+ T cells. (A) Naive CD4+ T cells were isolated from the spleens of wild-type and Runt–transgenic mice and stimulated with anti-CD3/anti-CD28 antibodies, and the culture supernatants were collected at the indicated times after stimulation. (B) The TCR-stimulated cells were cultured in the presence of IL-2 for 5 d, washed with fresh media, and restimulated via TCR. The culture supernatants were collected after 24 h. (C) The cells were cultured for four days in the Th2- or Th1-inducing condition. The cells were washed with fresh media and restimulated via TCR. The culture supernatants were collected after 24 h. (D) Same as C, but note the difference of scale used in C and here. The amounts of cytokines secreted into the supernatants were measured by ELISA and their averages and standard deviations are shown. Data are representative of four independent experiments.
Mentions: We previously established mouse lines that express a DNA binding domain of Runx1 from a transgene in a T-lineage-specific way (24). This Runt domain is known to function in a dominant interfering fashion against the Runx1 protein that is expressed endogenously in T lymphocytes. In the present study, we investigated the role of the Runx1 transcription factor in Th cell differentiation using these Runt-transgenic mice. Naive CD4+ T cells were isolated from the spleens of wild-type and Runt-transgenic mice and stimulated with anti-CD3/anti-CD28 antibodies. The culture supernatants were collected and the amount of cytokines secreted was measured by ELISA (Fig. 1 A). The Runt-transgenic cells produced five times as much IL-4 as the wild-type cells at 72 h after incubation. Similarly, the Runt-transgenic cells produced 1.7 and 3.4 times as much IL-5 and IL-10, respectively, as the wild-type cells. The Runt-transgenic cells thus exhibited enhanced production of Th2–type cytokines during an early phase of TCR activation. In contrast, secretion of IFN-γ and IL-2 from the Runt-transgenic cells was decreased slightly and markedly, respectively, compared with the wild-type cells. A similar result as that shown in Fig. 1 A was obtained when the TCR-stimulated cells were incubated in the presence of IL-2 for 5 d, washed with fresh media, and restimulated via TCR (Fig. 1 B). Under this neutral culture condition where neither IL-4 nor IL-12 was added to media, the Runt-transgenic cells secreted several fold more amount of Th2-type cytokines and less amount of Th1-type cytokines than the wild-type cells.

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