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Selective expression of an interleukin-12 receptor component by human T helper 1 cells.

Rogge L, Barberis-Maino L, Biffi M, Passini N, Presky DH, Gubler U, Sinigaglia F - J. Exp. Med. (1997)

Bottom Line: Here we show that the IL-12 receptor (IL-12R) beta 2 subunit, a recently cloned binding and signal transducing component of the IL-12R, is expressed on human Th1 but not Th2 clones and is induced during differentiation of human naive cells along the Th1 but not the Th2 pathway.IL-12 and type I but not type II interferons induce expression of the IL-12R beta 2 chain during in vitro T cell differentiation after antigen receptor triggering.The selective expression and regulation of the IL-12R beta 2 subunit may help to understand the basis of Th1/Th2 differentiation and may provide therapeutic options for altering the Th1/Th2 balance in several immuno-pathological conditions such as autoimmune diseases and allergies.

View Article: PubMed Central - PubMed

Affiliation: Roche Milano Ricerche, Italy.

ABSTRACT
Interleukin-12 (IL-12), a heterodimeric cytokine produced by activated monocytes and dendritic cells, plays a crucial role in regulating interferon (IFN)-gamma production and in the generation of IFN-gamma-producing T helper 1 (Th1) cells. Here we show that the IL-12 receptor (IL-12R) beta 2 subunit, a recently cloned binding and signal transducing component of the IL-12R, is expressed on human Th1 but not Th2 clones and is induced during differentiation of human naive cells along the Th1 but not the Th2 pathway. IL-12 and type I but not type II interferons induce expression of the IL-12R beta 2 chain during in vitro T cell differentiation after antigen receptor triggering. The selective expression and regulation of the IL-12R beta 2 subunit may help to understand the basis of Th1/Th2 differentiation and may provide therapeutic options for altering the Th1/Th2 balance in several immuno-pathological conditions such as autoimmune diseases and allergies.

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IL-12–dependent signaling in human Th1 and Th2 cell lines  and clones. IL-12 induces Stat4 phosphorylation in human Th1 but not  Th2 cells. Th1 and Th2 lines generated from cord blood lymphocytes as  in Fig. 1 A were harvested on day 10 after priming. Th1 (CB13) and Th2  (CB21) clones were harvested on day 13 after restimulation. 5 × 106 cells  were washed and incubated 15 min at 37°C in medium with or without  2 ng/ml IL-12 followed by the preparation of whole cell extracts and immunoprecipitation with anti-Stat4 antiserum. Precipitated proteins were  separated by SDS-PAGE (8%), transferred to nitrocellulose, and probed  with anti-phosphotyrosine antibody 4G10 (anti-P-Y, top). As a control for  Stat4 expression, blots were stripped and reprobed with anti-Stat4 antibodies (bottom).
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Figure 2: IL-12–dependent signaling in human Th1 and Th2 cell lines and clones. IL-12 induces Stat4 phosphorylation in human Th1 but not Th2 cells. Th1 and Th2 lines generated from cord blood lymphocytes as in Fig. 1 A were harvested on day 10 after priming. Th1 (CB13) and Th2 (CB21) clones were harvested on day 13 after restimulation. 5 × 106 cells were washed and incubated 15 min at 37°C in medium with or without 2 ng/ml IL-12 followed by the preparation of whole cell extracts and immunoprecipitation with anti-Stat4 antiserum. Precipitated proteins were separated by SDS-PAGE (8%), transferred to nitrocellulose, and probed with anti-phosphotyrosine antibody 4G10 (anti-P-Y, top). As a control for Stat4 expression, blots were stripped and reprobed with anti-Stat4 antibodies (bottom).

Mentions: To characterize the role of IL-12 and other cytokines in the differentiation of T helper cell subsets, we have generated Th1 and Th2 lines by stimulating human cord blood leukocytes with mitogen in the presence of IL-12, and neutralizing anti–IL-4 mAb or IL-4, and neutralizing anti–IL-12 mAb, respectively. This protocol allows the establishment of human T cell lines with strongly polarized cytokine production. Neonatal T cells primed under the Th1 conditions produced up to 2 ng/ml of IFN-γ and <0.2 ng/ml IL-4 upon restimulation with plate-bound anti-CD3 mAb, whereas neonatal T cells primed in the presence of IL-4 and anti–IL-12 mAb produced 1.2 ng/ml of IL-4 and <0.02 ng/ml IFN-γ (Fig. 1 A). Subsequent cloning of the Th1 and the Th2 lines resulted in a series of T cell clones with either a Th1 or a Th2 polarized cytokine profile, respectively (Fig. 1 B). Using a murine TCR transgenic system, Szabo et al. observed that IL-12 selectively activated the Janus kinase Jak2, and the signal transducers and activators of transcription Stat3 and Stat4 in Th1, but not in differentiated Th2 cells (30). To ascertain whether human Th1 and Th2 subsets also differ in IL-12 signaling, we examined Stat4 expression and tyrosine phosphorylation in response to IL-12 in Th1 and Th2 lines and clones. As shown in Fig. 2, IL-12 treatment induced tyrosine phosphorylation of Stat4 selectively in Th1 but not in Th2 cells, although both cell types expressed comparable amounts of the Stat4 protein (Fig. 2, bottom) required for IL-12 signaling (31, 32). Similarly, after IL-12 stimulation, Jak2 (33), which was also expressed at comparable levels in Th1 and Th2 cells, was phosphorylated only in Th1 cells (data not shown).


Selective expression of an interleukin-12 receptor component by human T helper 1 cells.

Rogge L, Barberis-Maino L, Biffi M, Passini N, Presky DH, Gubler U, Sinigaglia F - J. Exp. Med. (1997)

IL-12–dependent signaling in human Th1 and Th2 cell lines  and clones. IL-12 induces Stat4 phosphorylation in human Th1 but not  Th2 cells. Th1 and Th2 lines generated from cord blood lymphocytes as  in Fig. 1 A were harvested on day 10 after priming. Th1 (CB13) and Th2  (CB21) clones were harvested on day 13 after restimulation. 5 × 106 cells  were washed and incubated 15 min at 37°C in medium with or without  2 ng/ml IL-12 followed by the preparation of whole cell extracts and immunoprecipitation with anti-Stat4 antiserum. Precipitated proteins were  separated by SDS-PAGE (8%), transferred to nitrocellulose, and probed  with anti-phosphotyrosine antibody 4G10 (anti-P-Y, top). As a control for  Stat4 expression, blots were stripped and reprobed with anti-Stat4 antibodies (bottom).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2196163&req=5

Figure 2: IL-12–dependent signaling in human Th1 and Th2 cell lines and clones. IL-12 induces Stat4 phosphorylation in human Th1 but not Th2 cells. Th1 and Th2 lines generated from cord blood lymphocytes as in Fig. 1 A were harvested on day 10 after priming. Th1 (CB13) and Th2 (CB21) clones were harvested on day 13 after restimulation. 5 × 106 cells were washed and incubated 15 min at 37°C in medium with or without 2 ng/ml IL-12 followed by the preparation of whole cell extracts and immunoprecipitation with anti-Stat4 antiserum. Precipitated proteins were separated by SDS-PAGE (8%), transferred to nitrocellulose, and probed with anti-phosphotyrosine antibody 4G10 (anti-P-Y, top). As a control for Stat4 expression, blots were stripped and reprobed with anti-Stat4 antibodies (bottom).
Mentions: To characterize the role of IL-12 and other cytokines in the differentiation of T helper cell subsets, we have generated Th1 and Th2 lines by stimulating human cord blood leukocytes with mitogen in the presence of IL-12, and neutralizing anti–IL-4 mAb or IL-4, and neutralizing anti–IL-12 mAb, respectively. This protocol allows the establishment of human T cell lines with strongly polarized cytokine production. Neonatal T cells primed under the Th1 conditions produced up to 2 ng/ml of IFN-γ and <0.2 ng/ml IL-4 upon restimulation with plate-bound anti-CD3 mAb, whereas neonatal T cells primed in the presence of IL-4 and anti–IL-12 mAb produced 1.2 ng/ml of IL-4 and <0.02 ng/ml IFN-γ (Fig. 1 A). Subsequent cloning of the Th1 and the Th2 lines resulted in a series of T cell clones with either a Th1 or a Th2 polarized cytokine profile, respectively (Fig. 1 B). Using a murine TCR transgenic system, Szabo et al. observed that IL-12 selectively activated the Janus kinase Jak2, and the signal transducers and activators of transcription Stat3 and Stat4 in Th1, but not in differentiated Th2 cells (30). To ascertain whether human Th1 and Th2 subsets also differ in IL-12 signaling, we examined Stat4 expression and tyrosine phosphorylation in response to IL-12 in Th1 and Th2 lines and clones. As shown in Fig. 2, IL-12 treatment induced tyrosine phosphorylation of Stat4 selectively in Th1 but not in Th2 cells, although both cell types expressed comparable amounts of the Stat4 protein (Fig. 2, bottom) required for IL-12 signaling (31, 32). Similarly, after IL-12 stimulation, Jak2 (33), which was also expressed at comparable levels in Th1 and Th2 cells, was phosphorylated only in Th1 cells (data not shown).

Bottom Line: Here we show that the IL-12 receptor (IL-12R) beta 2 subunit, a recently cloned binding and signal transducing component of the IL-12R, is expressed on human Th1 but not Th2 clones and is induced during differentiation of human naive cells along the Th1 but not the Th2 pathway.IL-12 and type I but not type II interferons induce expression of the IL-12R beta 2 chain during in vitro T cell differentiation after antigen receptor triggering.The selective expression and regulation of the IL-12R beta 2 subunit may help to understand the basis of Th1/Th2 differentiation and may provide therapeutic options for altering the Th1/Th2 balance in several immuno-pathological conditions such as autoimmune diseases and allergies.

View Article: PubMed Central - PubMed

Affiliation: Roche Milano Ricerche, Italy.

ABSTRACT
Interleukin-12 (IL-12), a heterodimeric cytokine produced by activated monocytes and dendritic cells, plays a crucial role in regulating interferon (IFN)-gamma production and in the generation of IFN-gamma-producing T helper 1 (Th1) cells. Here we show that the IL-12 receptor (IL-12R) beta 2 subunit, a recently cloned binding and signal transducing component of the IL-12R, is expressed on human Th1 but not Th2 clones and is induced during differentiation of human naive cells along the Th1 but not the Th2 pathway. IL-12 and type I but not type II interferons induce expression of the IL-12R beta 2 chain during in vitro T cell differentiation after antigen receptor triggering. The selective expression and regulation of the IL-12R beta 2 subunit may help to understand the basis of Th1/Th2 differentiation and may provide therapeutic options for altering the Th1/Th2 balance in several immuno-pathological conditions such as autoimmune diseases and allergies.

Show MeSH
Related in: MedlinePlus