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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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Cytokine profiles of cord blood derived Th1 and Th2 lines  and of established human T cell clones. (A) IFN-γ and IL-4 production  by Th1 and Th2 lines generated from cord blood lymphocytes. Cells  were washed on day 10 after priming, 105 cells were restimulated in 96well round-bottom plates for 24 h with plate-bound anti-CD3 antibodies  (clone TR66; reference 19), to determine IFN-γ and IL-4 in culture supernatants by ELISA assays. The results shown are representative of five  independent experiments. (B) IFN-γ and IL-4 production by established  Th1 (CB13) and Th2 (CB21) clones.
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Figure 1: Cytokine profiles of cord blood derived Th1 and Th2 lines and of established human T cell clones. (A) IFN-γ and IL-4 production by Th1 and Th2 lines generated from cord blood lymphocytes. Cells were washed on day 10 after priming, 105 cells were restimulated in 96well round-bottom plates for 24 h with plate-bound anti-CD3 antibodies (clone TR66; reference 19), to determine IFN-γ and IL-4 in culture supernatants by ELISA assays. The results shown are representative of five independent experiments. (B) IFN-γ and IL-4 production by established Th1 (CB13) and Th2 (CB21) clones.

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)

Cytokine profiles of cord blood derived Th1 and Th2 lines  and of established human T cell clones. (A) IFN-γ and IL-4 production  by Th1 and Th2 lines generated from cord blood lymphocytes. Cells  were washed on day 10 after priming, 105 cells were restimulated in 96well round-bottom plates for 24 h with plate-bound anti-CD3 antibodies  (clone TR66; reference 19), to determine IFN-γ and IL-4 in culture supernatants by ELISA assays. The results shown are representative of five  independent experiments. (B) IFN-γ and IL-4 production by established  Th1 (CB13) and Th2 (CB21) clones.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2196163&req=5

Figure 1: Cytokine profiles of cord blood derived Th1 and Th2 lines and of established human T cell clones. (A) IFN-γ and IL-4 production by Th1 and Th2 lines generated from cord blood lymphocytes. Cells were washed on day 10 after priming, 105 cells were restimulated in 96well round-bottom plates for 24 h with plate-bound anti-CD3 antibodies (clone TR66; reference 19), to determine IFN-γ and IL-4 in culture supernatants by ELISA assays. The results shown are representative of five independent experiments. (B) IFN-γ and IL-4 production by established Th1 (CB13) and Th2 (CB21) clones.
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