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Regulation of the interleukin (IL)-12R beta 2 subunit expression in developing T helper 1 (Th1) and Th2 cells.

Szabo SJ, Dighe AS, Gubler U, Murphy KM - J. Exp. Med. (1997)

Bottom Line: Importantly, IL-4 and IFN-gamma were found to significantly modify IL-12 receptor beta 2 expression after T cell activation.IFN-gamma treatment of early developing Th2 cells maintained IL-12R beta 2 expression and restored the ability of these cells to functionally respond to IL-12, but did not directly inhibit IL-4 or induce IFN-gamma production.Controlling the expression of the IL-12R beta 2 subunit could be an important therapeutic target for the redirection of ongoing Th cell responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.

ABSTRACT
The developmental commitment to a T helper 1 (Th1)- or Th2-type response can significantly influence host immunity to pathogens. Extinction of the IL-12 signaling pathway during early Th2 development provides a mechanism that allows stable phenotype commitment. In this report we demonstrate that extinction of IL-12 signaling in early Th2 cells results from a selective loss of IL-12 receptor (IL-12R) beta 2 subunit expression. To determine the basis for this selective loss, we examined IL-12R beta 2 subunit expression during Th cell development in response to T cell treatment with different cytokines. IL-12R beta 2 is not expressed by naive resting CD4+ T cells, but is induced upon antigen activation through the T cell receptor. Importantly, IL-4 and IFN-gamma were found to significantly modify IL-12 receptor beta 2 expression after T cell activation. IL-4 inhibited IL-12R beta 2 expression leading to the loss of IL-12 signaling, providing an important point of regulation to promote commitment to the Th2 pathway. IFN-gamma treatment of early developing Th2 cells maintained IL-12R beta 2 expression and restored the ability of these cells to functionally respond to IL-12, but did not directly inhibit IL-4 or induce IFN-gamma production. Thus, IFN-gamma may prevent early Th cells from premature commitment to the Th2 pathway. Controlling the expression of the IL-12R beta 2 subunit could be an important therapeutic target for the redirection of ongoing Th cell responses.

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IL-12R β2 subunit mRNA is detected in Th1 but not Th2 cells. (A) Naive CD4+ T cells were purified by FACS® from unimmunized  DO11.10 TCR-transgenic mice as described in Materials and Methods (5), activated with OVA peptide and APCs under either Th1- or Th2-inducing conditions and allowed to develop for 7 days. On day 7, the Th1 and Th2 cells were washed, restimulated and allowed to proliferate for 7 or 9 d when cells  were harvested and total cellular RNA was isolated. As tissue controls, total cellular RNA was isolated from the B cell hybridoma TA3 and the fibroblast  cell line L929. Northern blot analysis was performed using as probes the full-length murine IL-12R β2 subunit cDNA (top), the full-length murine IL-12R  β1 subunit cDNA (middle), and the GAPDH cDNA (bottom). (B and C). Total cellular RNA was isolated from naive T cells after purification by FACS®.  Naive T cells isolated by cell sorting were activated to induce Th1 or Th2 development, and harvested on days 3, 5, and 7 after primary antigen activation. Total cellular RNA was examined by Northern analysis as described above for IL-12R β2 subunit cDNA (top), the IL-12R β1 subunit cDNA (middle), or pHE7 cDNA (bottom).
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Figure 1: IL-12R β2 subunit mRNA is detected in Th1 but not Th2 cells. (A) Naive CD4+ T cells were purified by FACS® from unimmunized DO11.10 TCR-transgenic mice as described in Materials and Methods (5), activated with OVA peptide and APCs under either Th1- or Th2-inducing conditions and allowed to develop for 7 days. On day 7, the Th1 and Th2 cells were washed, restimulated and allowed to proliferate for 7 or 9 d when cells were harvested and total cellular RNA was isolated. As tissue controls, total cellular RNA was isolated from the B cell hybridoma TA3 and the fibroblast cell line L929. Northern blot analysis was performed using as probes the full-length murine IL-12R β2 subunit cDNA (top), the full-length murine IL-12R β1 subunit cDNA (middle), and the GAPDH cDNA (bottom). (B and C). Total cellular RNA was isolated from naive T cells after purification by FACS®. Naive T cells isolated by cell sorting were activated to induce Th1 or Th2 development, and harvested on days 3, 5, and 7 after primary antigen activation. Total cellular RNA was examined by Northern analysis as described above for IL-12R β2 subunit cDNA (top), the IL-12R β1 subunit cDNA (middle), or pHE7 cDNA (bottom).

Mentions: Mel-14hi CD4+ T cells from DO11.10 TCRtransgenic mice were purified and activated with antigen in the presence of IL-12 and antibodies to IL-4, or IL-4 and antibodies to IL-12, for 7 d to generate polarized Th1 or Th2 populations (17). Subsequently, IL-12R expression was analyzed by Northern blotting on day 7 and 9 after secondary activation (Fig. 1 A). Th1 cells expressed the IL-12R β2 mRNA at high levels on these days. In contrast, IL-12R β2 mRNA was completely absent in Th2 cells harvested at these same time points. As we previously observed (17), IL-12R β1 mRNA was present in both Th1 and Th2 cells (Fig. 1 A, top).


Regulation of the interleukin (IL)-12R beta 2 subunit expression in developing T helper 1 (Th1) and Th2 cells.

Szabo SJ, Dighe AS, Gubler U, Murphy KM - J. Exp. Med. (1997)

IL-12R β2 subunit mRNA is detected in Th1 but not Th2 cells. (A) Naive CD4+ T cells were purified by FACS® from unimmunized  DO11.10 TCR-transgenic mice as described in Materials and Methods (5), activated with OVA peptide and APCs under either Th1- or Th2-inducing conditions and allowed to develop for 7 days. On day 7, the Th1 and Th2 cells were washed, restimulated and allowed to proliferate for 7 or 9 d when cells  were harvested and total cellular RNA was isolated. As tissue controls, total cellular RNA was isolated from the B cell hybridoma TA3 and the fibroblast  cell line L929. Northern blot analysis was performed using as probes the full-length murine IL-12R β2 subunit cDNA (top), the full-length murine IL-12R  β1 subunit cDNA (middle), and the GAPDH cDNA (bottom). (B and C). Total cellular RNA was isolated from naive T cells after purification by FACS®.  Naive T cells isolated by cell sorting were activated to induce Th1 or Th2 development, and harvested on days 3, 5, and 7 after primary antigen activation. Total cellular RNA was examined by Northern analysis as described above for IL-12R β2 subunit cDNA (top), the IL-12R β1 subunit cDNA (middle), or pHE7 cDNA (bottom).
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Related In: Results  -  Collection

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Figure 1: IL-12R β2 subunit mRNA is detected in Th1 but not Th2 cells. (A) Naive CD4+ T cells were purified by FACS® from unimmunized DO11.10 TCR-transgenic mice as described in Materials and Methods (5), activated with OVA peptide and APCs under either Th1- or Th2-inducing conditions and allowed to develop for 7 days. On day 7, the Th1 and Th2 cells were washed, restimulated and allowed to proliferate for 7 or 9 d when cells were harvested and total cellular RNA was isolated. As tissue controls, total cellular RNA was isolated from the B cell hybridoma TA3 and the fibroblast cell line L929. Northern blot analysis was performed using as probes the full-length murine IL-12R β2 subunit cDNA (top), the full-length murine IL-12R β1 subunit cDNA (middle), and the GAPDH cDNA (bottom). (B and C). Total cellular RNA was isolated from naive T cells after purification by FACS®. Naive T cells isolated by cell sorting were activated to induce Th1 or Th2 development, and harvested on days 3, 5, and 7 after primary antigen activation. Total cellular RNA was examined by Northern analysis as described above for IL-12R β2 subunit cDNA (top), the IL-12R β1 subunit cDNA (middle), or pHE7 cDNA (bottom).
Mentions: Mel-14hi CD4+ T cells from DO11.10 TCRtransgenic mice were purified and activated with antigen in the presence of IL-12 and antibodies to IL-4, or IL-4 and antibodies to IL-12, for 7 d to generate polarized Th1 or Th2 populations (17). Subsequently, IL-12R expression was analyzed by Northern blotting on day 7 and 9 after secondary activation (Fig. 1 A). Th1 cells expressed the IL-12R β2 mRNA at high levels on these days. In contrast, IL-12R β2 mRNA was completely absent in Th2 cells harvested at these same time points. As we previously observed (17), IL-12R β1 mRNA was present in both Th1 and Th2 cells (Fig. 1 A, top).

Bottom Line: Importantly, IL-4 and IFN-gamma were found to significantly modify IL-12 receptor beta 2 expression after T cell activation.IFN-gamma treatment of early developing Th2 cells maintained IL-12R beta 2 expression and restored the ability of these cells to functionally respond to IL-12, but did not directly inhibit IL-4 or induce IFN-gamma production.Controlling the expression of the IL-12R beta 2 subunit could be an important therapeutic target for the redirection of ongoing Th cell responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.

ABSTRACT
The developmental commitment to a T helper 1 (Th1)- or Th2-type response can significantly influence host immunity to pathogens. Extinction of the IL-12 signaling pathway during early Th2 development provides a mechanism that allows stable phenotype commitment. In this report we demonstrate that extinction of IL-12 signaling in early Th2 cells results from a selective loss of IL-12 receptor (IL-12R) beta 2 subunit expression. To determine the basis for this selective loss, we examined IL-12R beta 2 subunit expression during Th cell development in response to T cell treatment with different cytokines. IL-12R beta 2 is not expressed by naive resting CD4+ T cells, but is induced upon antigen activation through the T cell receptor. Importantly, IL-4 and IFN-gamma were found to significantly modify IL-12 receptor beta 2 expression after T cell activation. IL-4 inhibited IL-12R beta 2 expression leading to the loss of IL-12 signaling, providing an important point of regulation to promote commitment to the Th2 pathway. IFN-gamma treatment of early developing Th2 cells maintained IL-12R beta 2 expression and restored the ability of these cells to functionally respond to IL-12, but did not directly inhibit IL-4 or induce IFN-gamma production. Thus, IFN-gamma may prevent early Th cells from premature commitment to the Th2 pathway. Controlling the expression of the IL-12R beta 2 subunit could be an important therapeutic target for the redirection of ongoing Th cell responses.

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