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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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Regulation of IL-12R β2 subunit mRNA expression by  IFN-γ. FACS®-sorted naive CD4+ DO11.10 T cells were cultured with  OVA peptide and irradiated BALB/c splenocytes under conditions indicated. On day 5 after primary antigen activation total cellular RNA was  isolated from the developing Th cells. Sequential 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).
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Figure 5: Regulation of IL-12R β2 subunit mRNA expression by IFN-γ. FACS®-sorted naive CD4+ DO11.10 T cells were cultured with OVA peptide and irradiated BALB/c splenocytes under conditions indicated. On day 5 after primary antigen activation total cellular RNA was isolated from the developing Th cells. Sequential 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).

Mentions: To test whether IFN-γ–induced restoration of IL-12 signaling involved induction of the IL-12R β2 subunit, we performed Northern blot analysis of Th cells derived under the various conditions described in Fig. 4. T cells activated in the presence of both IL-4 and IL-12 expressed the IL-12R β2 mRNA (Fig. 5, lane 4). This expression was dependent on endogenous IFN-γ production, since neutralization of IFN-γ in the primary culture led to the disappearance of IL-12R β2 mRNA (Fig. 5, lane 3). T cells arising from stimulation in the presence of IL-4, anti–IL-12 mAb and exogenous IFN-γ added during primary activation expressed high levels of the IL-12R β2 mRNA (Fig. 5, lane 6). Addition of anti–IFN-γ mAb blocked expression of IL-12R β2 mRNA (Fig. 5, lane 5). Th1 cells, arising from stimulation in the presence of IL-12 plus anti–IL-4 mAb, expressed IL-12R β2 mRNA independently of IFN-γ (Fig. 5, lanes 1 and 2). Thus, in each case, IL-12-inducible Stat4 phosphorylation (Fig. 4) correlated with expression of the IL-12R β2 mRNA (Fig. 5).


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)

Regulation of IL-12R β2 subunit mRNA expression by  IFN-γ. FACS®-sorted naive CD4+ DO11.10 T cells were cultured with  OVA peptide and irradiated BALB/c splenocytes under conditions indicated. On day 5 after primary antigen activation total cellular RNA was  isolated from the developing Th cells. Sequential 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).
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Related In: Results  -  Collection

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Figure 5: Regulation of IL-12R β2 subunit mRNA expression by IFN-γ. FACS®-sorted naive CD4+ DO11.10 T cells were cultured with OVA peptide and irradiated BALB/c splenocytes under conditions indicated. On day 5 after primary antigen activation total cellular RNA was isolated from the developing Th cells. Sequential 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).
Mentions: To test whether IFN-γ–induced restoration of IL-12 signaling involved induction of the IL-12R β2 subunit, we performed Northern blot analysis of Th cells derived under the various conditions described in Fig. 4. T cells activated in the presence of both IL-4 and IL-12 expressed the IL-12R β2 mRNA (Fig. 5, lane 4). This expression was dependent on endogenous IFN-γ production, since neutralization of IFN-γ in the primary culture led to the disappearance of IL-12R β2 mRNA (Fig. 5, lane 3). T cells arising from stimulation in the presence of IL-4, anti–IL-12 mAb and exogenous IFN-γ added during primary activation expressed high levels of the IL-12R β2 mRNA (Fig. 5, lane 6). Addition of anti–IFN-γ mAb blocked expression of IL-12R β2 mRNA (Fig. 5, lane 5). Th1 cells, arising from stimulation in the presence of IL-12 plus anti–IL-4 mAb, expressed IL-12R β2 mRNA independently of IFN-γ (Fig. 5, lanes 1 and 2). Thus, in each case, IL-12-inducible Stat4 phosphorylation (Fig. 4) correlated with expression of the IL-12R β2 mRNA (Fig. 5).

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.

Show MeSH