Limits...
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.

Show MeSH
Effect of Runx1 overexpression and Runt-transgene on GATA3 expression. (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 and reactivated via TCR. The GFP+ population was sorted as shown in the top panel, and RNA was extracted and processed for Northern blot analysis as shown in the lower panel. (B) Naive CD4+ T cells were isolated from the spleens of wild-type and Runt-transgenic mice, and stimulated with anti-CD3/anti-CD28 antibodies. In case, either anti–IL-4 antibody or IL-4 or IL-12 was added to the culture medium as indicated. RNA was extracted and processed for Northern blot analysis. Relative intensity of GATA3 and T-bet transcripts that were normalized by that of G3PDH are shown below the lanes. Data are representative of two independent experiments.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2196077&req=5

fig5: Effect of Runx1 overexpression and Runt-transgene on GATA3 expression. (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 and reactivated via TCR. The GFP+ population was sorted as shown in the top panel, and RNA was extracted and processed for Northern blot analysis as shown in the lower panel. (B) Naive CD4+ T cells were isolated from the spleens of wild-type and Runt-transgenic mice, and stimulated with anti-CD3/anti-CD28 antibodies. In case, either anti–IL-4 antibody or IL-4 or IL-12 was added to the culture medium as indicated. RNA was extracted and processed for Northern blot analysis. Relative intensity of GATA3 and T-bet transcripts that were normalized by that of G3PDH are shown below the lanes. Data are representative of two independent experiments.

Mentions: The GATA3 transcription factor is located downstream of IL-4R signaling pathway and regulates the commitment of cells toward the Th2 lineage (11). We therefore examined whether overexpression of Runx1 affects GATA3 expression. The RNA was extracted from the GFP+ fractions and processed for Northern blot analysis (Fig. 5 A). In the pMX-infected cells, some amount of GATA3 transcript was detected (lane 1), whereas almost no GATA3 transcript was detected in the pMX-Runx1–infected cells (lane 3). Furthermore, addition of an excess amount of IL-4 into the medium induced higher expression of GATA3 in the pMX-infected cells (lane 2), but could not restore GATA3 expression in the Runx1-overexpressing cells (lane 4). The amount of T-bet, a Th1-specific transcription factor (32), decreased in response to the addition of IL-4 in both the Runx1-overexpressing and pMX-infected cells (lanes 2 and 4). Thus, Runx1 can strongly repress the induction of GATA3 expression and the inability of Runx1-overexpressing cells to differentiate into the Th2 lineage is likely due to this paucity of GATA3 induction.


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 Runx1 overexpression and Runt-transgene on GATA3 expression. (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 and reactivated via TCR. The GFP+ population was sorted as shown in the top panel, and RNA was extracted and processed for Northern blot analysis as shown in the lower panel. (B) Naive CD4+ T cells were isolated from the spleens of wild-type and Runt-transgenic mice, and stimulated with anti-CD3/anti-CD28 antibodies. In case, either anti–IL-4 antibody or IL-4 or IL-12 was added to the culture medium as indicated. RNA was extracted and processed for Northern blot analysis. Relative intensity of GATA3 and T-bet transcripts that were normalized by that of G3PDH are shown below the lanes. Data are representative of two independent experiments.
© Copyright Policy
Related In: Results  -  Collection

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

fig5: Effect of Runx1 overexpression and Runt-transgene on GATA3 expression. (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 and reactivated via TCR. The GFP+ population was sorted as shown in the top panel, and RNA was extracted and processed for Northern blot analysis as shown in the lower panel. (B) Naive CD4+ T cells were isolated from the spleens of wild-type and Runt-transgenic mice, and stimulated with anti-CD3/anti-CD28 antibodies. In case, either anti–IL-4 antibody or IL-4 or IL-12 was added to the culture medium as indicated. RNA was extracted and processed for Northern blot analysis. Relative intensity of GATA3 and T-bet transcripts that were normalized by that of G3PDH are shown below the lanes. Data are representative of two independent experiments.
Mentions: The GATA3 transcription factor is located downstream of IL-4R signaling pathway and regulates the commitment of cells toward the Th2 lineage (11). We therefore examined whether overexpression of Runx1 affects GATA3 expression. The RNA was extracted from the GFP+ fractions and processed for Northern blot analysis (Fig. 5 A). In the pMX-infected cells, some amount of GATA3 transcript was detected (lane 1), whereas almost no GATA3 transcript was detected in the pMX-Runx1–infected cells (lane 3). Furthermore, addition of an excess amount of IL-4 into the medium induced higher expression of GATA3 in the pMX-infected cells (lane 2), but could not restore GATA3 expression in the Runx1-overexpressing cells (lane 4). The amount of T-bet, a Th1-specific transcription factor (32), decreased in response to the addition of IL-4 in both the Runx1-overexpressing and pMX-infected cells (lanes 2 and 4). Thus, Runx1 can strongly repress the induction of GATA3 expression and the inability of Runx1-overexpressing cells to differentiate into the Th2 lineage is likely due to this paucity of GATA3 induction.

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