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Interferon gamma signaling alters the function of T helper type 1 cells.

Tau GZ, von der Weid T, Lu B, Cowan S, Kvatyuk M, Pernis A, Cattoretti G, Braunstein NS, Coffman RL, Rothman PB - J. Exp. Med. (2000)

Bottom Line: For example, Th2 cells express both chains of the interferon gamma receptor (IFN-gammaR), whereas Th1 cells do not express the second chain of the IFN-gammaR (IFN-gammaR2) and are therefore unresponsive to IFN-gamma.Together, these data suggest that the sustained expression of IFN-gammaR2 inhibits Th1 differentiation and function.Therefore, the acquisition of an IFN-gamma-unresponsive phenotype in Th1 cells plays a crucial role in the development and function of these cells.

View Article: PubMed Central - PubMed

Affiliation: Integrated Program in Cell, Molecular and Biophysical Studies, Columbia University, New York, New York 10032, USA.

ABSTRACT
One mechanism regulating the ability of different subsets of T helper (Th) cells to respond to cytokines is the differential expression of cytokine receptors. For example, Th2 cells express both chains of the interferon gamma receptor (IFN-gammaR), whereas Th1 cells do not express the second chain of the IFN-gammaR (IFN-gammaR2) and are therefore unresponsive to IFN-gamma. To determine whether the regulation of IFN-gammaR2 expression, and therefore IFN-gamma responsiveness, is important for the differentiation of naive CD4(+) T cells into Th1 cells or for Th1 effector function, we generated mice in which transgenic (TG) expression of IFN-gammaR2 is controlled by the CD2 promoter and enhancer. CD4(+) T cells from IFN-gammaR2 TG mice exhibit impaired Th1 polarization potential in vitro. TG mice also display several defects in Th1-dependent immunity in vivo, including attenuated delayed-type hypersensitivity responses and decreased antigen-specific IFN-gamma production. In addition, TG mice mount impaired Th1 responses against Leishmania major, as manifested by increased parasitemia and more severe lesions than their wild-type littermates. Together, these data suggest that the sustained expression of IFN-gammaR2 inhibits Th1 differentiation and function. Therefore, the acquisition of an IFN-gamma-unresponsive phenotype in Th1 cells plays a crucial role in the development and function of these cells.

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The generation of IFN-γR2 TG mice, transgene expression, and IFN-γ responsiveness. (A) A diagram of the CD2 promoter IFN-γR2-CD2 enhancer construct used to generate IFN-γR2 TG mice. The template used to synthesize the antisense mRNA probe used in RNase protection assays is diagrammed above the TG construct. UTR, untranslated region. (B) The expression of the endogenous (End.) and TG alleles of IFN-γR2 in different organs was detected by RNase protection. (C) IFN-γR2 gene expression in Th1 and Th2 clones was detected by reverse transcription PCR and visualized by autoradiography. T, transgenic; E, endogenous. (D) The detection by electrophoretic mobility shift assays of activated STAT complexes by Th1 and Th2 clones in response to treatment with IL-4 or IFN-γ. All experiments were repeated three times with similar results.
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Figure 1: The generation of IFN-γR2 TG mice, transgene expression, and IFN-γ responsiveness. (A) A diagram of the CD2 promoter IFN-γR2-CD2 enhancer construct used to generate IFN-γR2 TG mice. The template used to synthesize the antisense mRNA probe used in RNase protection assays is diagrammed above the TG construct. UTR, untranslated region. (B) The expression of the endogenous (End.) and TG alleles of IFN-γR2 in different organs was detected by RNase protection. (C) IFN-γR2 gene expression in Th1 and Th2 clones was detected by reverse transcription PCR and visualized by autoradiography. T, transgenic; E, endogenous. (D) The detection by electrophoretic mobility shift assays of activated STAT complexes by Th1 and Th2 clones in response to treatment with IL-4 or IFN-γ. All experiments were repeated three times with similar results.

Mentions: Ribonuclease protection assay was performed as described previously 17. In brief, a SacI-ClaI fragment, diagrammed in Fig. 1 A, from the CD2-IFN-γR2 construct was subcloned into the pBluescript II SK+ phagemid (Stratagene). Radiolabeled antisense mRNA probe was synthesized using an RNA transcription kit (Stratagene). Radiolabled probe (105 cpm) was hybridized with 30 μg of tissue RNA in 80% deionized formamide at 42°C overnight. Samples were then digested with RNases A and T1 and run on an 8 M urea-6% acrylamide gel.


Interferon gamma signaling alters the function of T helper type 1 cells.

Tau GZ, von der Weid T, Lu B, Cowan S, Kvatyuk M, Pernis A, Cattoretti G, Braunstein NS, Coffman RL, Rothman PB - J. Exp. Med. (2000)

The generation of IFN-γR2 TG mice, transgene expression, and IFN-γ responsiveness. (A) A diagram of the CD2 promoter IFN-γR2-CD2 enhancer construct used to generate IFN-γR2 TG mice. The template used to synthesize the antisense mRNA probe used in RNase protection assays is diagrammed above the TG construct. UTR, untranslated region. (B) The expression of the endogenous (End.) and TG alleles of IFN-γR2 in different organs was detected by RNase protection. (C) IFN-γR2 gene expression in Th1 and Th2 clones was detected by reverse transcription PCR and visualized by autoradiography. T, transgenic; E, endogenous. (D) The detection by electrophoretic mobility shift assays of activated STAT complexes by Th1 and Th2 clones in response to treatment with IL-4 or IFN-γ. All experiments were repeated three times with similar results.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2193318&req=5

Figure 1: The generation of IFN-γR2 TG mice, transgene expression, and IFN-γ responsiveness. (A) A diagram of the CD2 promoter IFN-γR2-CD2 enhancer construct used to generate IFN-γR2 TG mice. The template used to synthesize the antisense mRNA probe used in RNase protection assays is diagrammed above the TG construct. UTR, untranslated region. (B) The expression of the endogenous (End.) and TG alleles of IFN-γR2 in different organs was detected by RNase protection. (C) IFN-γR2 gene expression in Th1 and Th2 clones was detected by reverse transcription PCR and visualized by autoradiography. T, transgenic; E, endogenous. (D) The detection by electrophoretic mobility shift assays of activated STAT complexes by Th1 and Th2 clones in response to treatment with IL-4 or IFN-γ. All experiments were repeated three times with similar results.
Mentions: Ribonuclease protection assay was performed as described previously 17. In brief, a SacI-ClaI fragment, diagrammed in Fig. 1 A, from the CD2-IFN-γR2 construct was subcloned into the pBluescript II SK+ phagemid (Stratagene). Radiolabeled antisense mRNA probe was synthesized using an RNA transcription kit (Stratagene). Radiolabled probe (105 cpm) was hybridized with 30 μg of tissue RNA in 80% deionized formamide at 42°C overnight. Samples were then digested with RNases A and T1 and run on an 8 M urea-6% acrylamide gel.

Bottom Line: For example, Th2 cells express both chains of the interferon gamma receptor (IFN-gammaR), whereas Th1 cells do not express the second chain of the IFN-gammaR (IFN-gammaR2) and are therefore unresponsive to IFN-gamma.Together, these data suggest that the sustained expression of IFN-gammaR2 inhibits Th1 differentiation and function.Therefore, the acquisition of an IFN-gamma-unresponsive phenotype in Th1 cells plays a crucial role in the development and function of these cells.

View Article: PubMed Central - PubMed

Affiliation: Integrated Program in Cell, Molecular and Biophysical Studies, Columbia University, New York, New York 10032, USA.

ABSTRACT
One mechanism regulating the ability of different subsets of T helper (Th) cells to respond to cytokines is the differential expression of cytokine receptors. For example, Th2 cells express both chains of the interferon gamma receptor (IFN-gammaR), whereas Th1 cells do not express the second chain of the IFN-gammaR (IFN-gammaR2) and are therefore unresponsive to IFN-gamma. To determine whether the regulation of IFN-gammaR2 expression, and therefore IFN-gamma responsiveness, is important for the differentiation of naive CD4(+) T cells into Th1 cells or for Th1 effector function, we generated mice in which transgenic (TG) expression of IFN-gammaR2 is controlled by the CD2 promoter and enhancer. CD4(+) T cells from IFN-gammaR2 TG mice exhibit impaired Th1 polarization potential in vitro. TG mice also display several defects in Th1-dependent immunity in vivo, including attenuated delayed-type hypersensitivity responses and decreased antigen-specific IFN-gamma production. In addition, TG mice mount impaired Th1 responses against Leishmania major, as manifested by increased parasitemia and more severe lesions than their wild-type littermates. Together, these data suggest that the sustained expression of IFN-gammaR2 inhibits Th1 differentiation and function. Therefore, the acquisition of an IFN-gamma-unresponsive phenotype in Th1 cells plays a crucial role in the development and function of these cells.

Show MeSH
Related in: MedlinePlus