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GATA-3 in human T cell helper type 2 development.

Skapenko A, Leipe J, Niesner U, Devriendt K, Beetz R, Radbruch A, Kalden JR, Lipsky PE, Schulze-Koops H - J. Exp. Med. (2004)

Bottom Line: CD4 T cells from GATA-3+/- individuals expressed significantly reduced levels of GATA-3, associated with markedly decreased T helper cell (Th)2 frequencies in vivo and in vitro.Moreover, Th2 cell-mediated effector functions, as assessed by serum levels of Th2-dependent immunoglobulins (Igs; IgG4, IgE), were dramatically decreased, whereas the Th1-dependent IgG1 was elevated compared with GATA-3+/+ controls.Concordant with these data, silencing of GATA-3 in GATA-3+/+ CD4 T cells with small interfering RNA significantly reduced Th2 cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Nikolaus Fiebiger Center for Molecular Medicine, Clinical Research Group III, University of Erlangen-Nuremberg, 91054, Germany.

ABSTRACT
The delineation of the in vivo role of GATA-3 in human T cell differentiation is a critical step in the understanding of molecular mechanisms directing human immune responses. We examined T cell differentiation and T cell-mediated effector functions in individuals lacking one functional GATA-3 allele. CD4 T cells from GATA-3+/- individuals expressed significantly reduced levels of GATA-3, associated with markedly decreased T helper cell (Th)2 frequencies in vivo and in vitro. Moreover, Th2 cell-mediated effector functions, as assessed by serum levels of Th2-dependent immunoglobulins (Igs; IgG4, IgE), were dramatically decreased, whereas the Th1-dependent IgG1 was elevated compared with GATA-3+/+ controls. Concordant with these data, silencing of GATA-3 in GATA-3+/+ CD4 T cells with small interfering RNA significantly reduced Th2 cell differentiation. Moreover, GATA-3 mRNA levels increased under Th2-inducing conditions and decreased under Th1-inducing conditions. Taken together, the data strongly suggest that GATA-3 is an important transcription factor in regulating human Th2 cell differentiation in vivo.

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CD4 T cells from GATA-3+/− individuals express reduced levels of GATA-3 associated with decreased Th2 cell differentiation and reduced Th2 effector functions. (A) Whole cell lysates of CD4 T cells from GATA-3+/− (individual no. 1) and GATA-3+/+ (control) individuals were analyzed by Western blotting with mAbs to GATA-3 and actin. One representative of three independent experiments with similar results is shown. (B) Freshly isolated memory CD4 T cells from GATA-3+/− individuals were stimulated in vitro with PMA and ionomycin and analyzed for cytoplasmic IL-4 (Th2) and IFN-γ (Th1). Dots indicate Th2 and Th1 cell frequencies of the individual GATA-3+/− individuals, whereas the shaded areas denote mean ± SD of 20 age-matched control individuals. (C) Freshly isolated naive T cells of GATA-3+/− individuals were primed in vitro under Th2-inducing conditions, stimulated with PMA and ionomycin, and assessed for cytoplasmic IL-4 and IFN-γ. Dots indicate Th2 and Th1 cell frequencies of two GATA-3+/− individuals. The third GATA-3+/− individual was not analyzed because of limited numbers of available cells. Shaded areas denote the normal range of control individuals. (D and E) Serum Ig levels of GATA-3+/− individuals. Shaded areas reflect the normal limits.
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fig1: CD4 T cells from GATA-3+/− individuals express reduced levels of GATA-3 associated with decreased Th2 cell differentiation and reduced Th2 effector functions. (A) Whole cell lysates of CD4 T cells from GATA-3+/− (individual no. 1) and GATA-3+/+ (control) individuals were analyzed by Western blotting with mAbs to GATA-3 and actin. One representative of three independent experiments with similar results is shown. (B) Freshly isolated memory CD4 T cells from GATA-3+/− individuals were stimulated in vitro with PMA and ionomycin and analyzed for cytoplasmic IL-4 (Th2) and IFN-γ (Th1). Dots indicate Th2 and Th1 cell frequencies of the individual GATA-3+/− individuals, whereas the shaded areas denote mean ± SD of 20 age-matched control individuals. (C) Freshly isolated naive T cells of GATA-3+/− individuals were primed in vitro under Th2-inducing conditions, stimulated with PMA and ionomycin, and assessed for cytoplasmic IL-4 and IFN-γ. Dots indicate Th2 and Th1 cell frequencies of two GATA-3+/− individuals. The third GATA-3+/− individual was not analyzed because of limited numbers of available cells. Shaded areas denote the normal range of control individuals. (D and E) Serum Ig levels of GATA-3+/− individuals. Shaded areas reflect the normal limits.

Mentions: Indications of protein functions in vivo have largely been derived from gene knockout approaches. However, the analysis of proteins critically involved in embryogenesis or early organ development is difficult to pursue if the target disruption of the particular gene results in the death of the embryo. To evaluate the role of GATA-3 in human Th2 cell differentiation in vivo, we analyzed three individuals lacking one functional GATA-3 allele (Table I). CD4 T cells from those individuals contained significantly less GATA-3 protein than CD4 T cells from controls (Fig. 1 A). Ex vivo analysis of cytokine production by CD4 memory T cells from the peripheral blood of the GATA-3+/− individuals revealed that the frequencies of IL-4–producing Th2 cells were at the lower end of or below the normal range (Fig. 1 B). In contrast, the frequencies of IFN-γ–producing Th1 cells were within the normal range (Fig. 1 B). The decreased Th2 cell frequencies in the peripheral blood of the GATA-3+/− individuals resulted in a shift of the Th2/Th1 balance, reflecting a Th1 bias in these individuals (Table II).


GATA-3 in human T cell helper type 2 development.

Skapenko A, Leipe J, Niesner U, Devriendt K, Beetz R, Radbruch A, Kalden JR, Lipsky PE, Schulze-Koops H - J. Exp. Med. (2004)

CD4 T cells from GATA-3+/− individuals express reduced levels of GATA-3 associated with decreased Th2 cell differentiation and reduced Th2 effector functions. (A) Whole cell lysates of CD4 T cells from GATA-3+/− (individual no. 1) and GATA-3+/+ (control) individuals were analyzed by Western blotting with mAbs to GATA-3 and actin. One representative of three independent experiments with similar results is shown. (B) Freshly isolated memory CD4 T cells from GATA-3+/− individuals were stimulated in vitro with PMA and ionomycin and analyzed for cytoplasmic IL-4 (Th2) and IFN-γ (Th1). Dots indicate Th2 and Th1 cell frequencies of the individual GATA-3+/− individuals, whereas the shaded areas denote mean ± SD of 20 age-matched control individuals. (C) Freshly isolated naive T cells of GATA-3+/− individuals were primed in vitro under Th2-inducing conditions, stimulated with PMA and ionomycin, and assessed for cytoplasmic IL-4 and IFN-γ. Dots indicate Th2 and Th1 cell frequencies of two GATA-3+/− individuals. The third GATA-3+/− individual was not analyzed because of limited numbers of available cells. Shaded areas denote the normal range of control individuals. (D and E) Serum Ig levels of GATA-3+/− individuals. Shaded areas reflect the normal limits.
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Related In: Results  -  Collection

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fig1: CD4 T cells from GATA-3+/− individuals express reduced levels of GATA-3 associated with decreased Th2 cell differentiation and reduced Th2 effector functions. (A) Whole cell lysates of CD4 T cells from GATA-3+/− (individual no. 1) and GATA-3+/+ (control) individuals were analyzed by Western blotting with mAbs to GATA-3 and actin. One representative of three independent experiments with similar results is shown. (B) Freshly isolated memory CD4 T cells from GATA-3+/− individuals were stimulated in vitro with PMA and ionomycin and analyzed for cytoplasmic IL-4 (Th2) and IFN-γ (Th1). Dots indicate Th2 and Th1 cell frequencies of the individual GATA-3+/− individuals, whereas the shaded areas denote mean ± SD of 20 age-matched control individuals. (C) Freshly isolated naive T cells of GATA-3+/− individuals were primed in vitro under Th2-inducing conditions, stimulated with PMA and ionomycin, and assessed for cytoplasmic IL-4 and IFN-γ. Dots indicate Th2 and Th1 cell frequencies of two GATA-3+/− individuals. The third GATA-3+/− individual was not analyzed because of limited numbers of available cells. Shaded areas denote the normal range of control individuals. (D and E) Serum Ig levels of GATA-3+/− individuals. Shaded areas reflect the normal limits.
Mentions: Indications of protein functions in vivo have largely been derived from gene knockout approaches. However, the analysis of proteins critically involved in embryogenesis or early organ development is difficult to pursue if the target disruption of the particular gene results in the death of the embryo. To evaluate the role of GATA-3 in human Th2 cell differentiation in vivo, we analyzed three individuals lacking one functional GATA-3 allele (Table I). CD4 T cells from those individuals contained significantly less GATA-3 protein than CD4 T cells from controls (Fig. 1 A). Ex vivo analysis of cytokine production by CD4 memory T cells from the peripheral blood of the GATA-3+/− individuals revealed that the frequencies of IL-4–producing Th2 cells were at the lower end of or below the normal range (Fig. 1 B). In contrast, the frequencies of IFN-γ–producing Th1 cells were within the normal range (Fig. 1 B). The decreased Th2 cell frequencies in the peripheral blood of the GATA-3+/− individuals resulted in a shift of the Th2/Th1 balance, reflecting a Th1 bias in these individuals (Table II).

Bottom Line: CD4 T cells from GATA-3+/- individuals expressed significantly reduced levels of GATA-3, associated with markedly decreased T helper cell (Th)2 frequencies in vivo and in vitro.Moreover, Th2 cell-mediated effector functions, as assessed by serum levels of Th2-dependent immunoglobulins (Igs; IgG4, IgE), were dramatically decreased, whereas the Th1-dependent IgG1 was elevated compared with GATA-3+/+ controls.Concordant with these data, silencing of GATA-3 in GATA-3+/+ CD4 T cells with small interfering RNA significantly reduced Th2 cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Nikolaus Fiebiger Center for Molecular Medicine, Clinical Research Group III, University of Erlangen-Nuremberg, 91054, Germany.

ABSTRACT
The delineation of the in vivo role of GATA-3 in human T cell differentiation is a critical step in the understanding of molecular mechanisms directing human immune responses. We examined T cell differentiation and T cell-mediated effector functions in individuals lacking one functional GATA-3 allele. CD4 T cells from GATA-3+/- individuals expressed significantly reduced levels of GATA-3, associated with markedly decreased T helper cell (Th)2 frequencies in vivo and in vitro. Moreover, Th2 cell-mediated effector functions, as assessed by serum levels of Th2-dependent immunoglobulins (Igs; IgG4, IgE), were dramatically decreased, whereas the Th1-dependent IgG1 was elevated compared with GATA-3+/+ controls. Concordant with these data, silencing of GATA-3 in GATA-3+/+ CD4 T cells with small interfering RNA significantly reduced Th2 cell differentiation. Moreover, GATA-3 mRNA levels increased under Th2-inducing conditions and decreased under Th1-inducing conditions. Taken together, the data strongly suggest that GATA-3 is an important transcription factor in regulating human Th2 cell differentiation in vivo.

Show MeSH
Related in: MedlinePlus