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Regulation of interleukin (IL)-12 receptor beta2 subunit expression by endogenous IL-12: a critical step in the differentiation of pathogenic autoreactive T cells.

Chang JT, Shevach EM, Segal BM - J. Exp. Med. (1999)

Bottom Line: Defective expression was not secondary to the production of suppressive cytokines, but to a failure of B10.S MBP-specific T cells to upregulate CD40 ligand expression and to induce the production of IL-12.IL-12Rbeta2 expression as well as encephalitogenicity of these cells could be restored by the addition of IL-12.These results suggest that the development of immunotherapies that target the IL-12Rbeta2 subunit may be useful for the treatment of autoimmune diseases.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.

ABSTRACT
The interleukin (IL)-12 receptor (R)beta2 subunit is the critical molecule involved in maintaining IL-12 responsiveness and controlling T helper cell type 1 lineage commitment. We demonstrate that IL-12 and interferon (IFN)-gamma play separate, but complementary, roles in regulating IL-12Rbeta2 expression on antigen-specific CD4(+) T cells. These results are consistent with our previous observation that IL-12 can promote autoimmune disease through IFN-gamma-independent as well as -dependent pathways. Therefore, we compared the induction of IL-12 by, and the expression of the IL-12Rbeta2 subunit on, myelin basic protein (MBP)-specific T cells from experimental allergic encephalomyelitis (EAE)-susceptible SJL (H-2(s)) mice and from EAE- resistant B10.S mice (H-2(s)). B10.S mice had an antigen-specific defect in their capacity to upregulate the IL-12Rbeta2 subunit. Defective expression was not secondary to the production of suppressive cytokines, but to a failure of B10.S MBP-specific T cells to upregulate CD40 ligand expression and to induce the production of IL-12. IL-12Rbeta2 expression as well as encephalitogenicity of these cells could be restored by the addition of IL-12. These results suggest that the development of immunotherapies that target the IL-12Rbeta2 subunit may be useful for the treatment of autoimmune diseases.

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Maintenance of IL-12Rβ2  mRNA expression is dependent on the presence of IL-12 and IFN-γ. (A) LN cells from  B10.S mice coimmunized with MBP/NP  were removed on day 12 after immunization  and cultured for 3 d in the presence or absence of MBP with or without IL-12 (2 or  0.2 ng/ml). Total RNA was extracted, and  Northern blot analysis was performed for  IL-12Rβ2 subunit and β-actin. Results are  representative of three experiments each using LN cells pooled from five or more mice. (B) LN cells from B10.S mice coimmunized with MBP/NP were removed on day 12 after immunization and  cultured for 3 d in the presence of NP with or without anti–IL-12 (10 μg /ml) or anti–IFN-γ (10 μg/ml). Total RNA was extracted, and Northern blot  analysis was performed for IL-12Rβ2 subunit and β-actin. Results are representative of three experiments each using LN cells pooled from five or more  mice. (C) LN cells from SJL mice coimmunized with MBP/NP were removed on day 12 after immunization and cultured for 3 d in the presence of MBP  with or without anti–IL-12 (10 μg/ml) or anti–IFN-γ (10 μg/ml). Total RNA was extracted, and Northern blot analysis was performed for IL-12Rβ2  subunit and β-actin. Results are representative of three experiments each using LN cells pooled from five or more mice.
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Figure 5: Maintenance of IL-12Rβ2 mRNA expression is dependent on the presence of IL-12 and IFN-γ. (A) LN cells from B10.S mice coimmunized with MBP/NP were removed on day 12 after immunization and cultured for 3 d in the presence or absence of MBP with or without IL-12 (2 or 0.2 ng/ml). Total RNA was extracted, and Northern blot analysis was performed for IL-12Rβ2 subunit and β-actin. Results are representative of three experiments each using LN cells pooled from five or more mice. (B) LN cells from B10.S mice coimmunized with MBP/NP were removed on day 12 after immunization and cultured for 3 d in the presence of NP with or without anti–IL-12 (10 μg /ml) or anti–IFN-γ (10 μg/ml). Total RNA was extracted, and Northern blot analysis was performed for IL-12Rβ2 subunit and β-actin. Results are representative of three experiments each using LN cells pooled from five or more mice. (C) LN cells from SJL mice coimmunized with MBP/NP were removed on day 12 after immunization and cultured for 3 d in the presence of MBP with or without anti–IL-12 (10 μg/ml) or anti–IFN-γ (10 μg/ml). Total RNA was extracted, and Northern blot analysis was performed for IL-12Rβ2 subunit and β-actin. Results are representative of three experiments each using LN cells pooled from five or more mice.

Mentions: We have previously demonstrated that the addition of pharmacological concentrations of IL-12 to cultures of B10.S MBP-reactive LN cells restored their ability to produce IFN-γ and converted them into encephalitogenic effectors (28). Indeed, the addition of exogenous IL-12 restored the ability of the B10.S MBP-reactive LN cells to upregulate the expression of IL-12Rβ2 subunit mRNA in an antigen-specific, dose-dependent manner (Fig. 5 A). Although the addition of IFN-γ failed to restore the ability of these cells to produce IFN-γ upon secondary stimulation (28), modest induction of IL-12Rβ2 subunit expression was seen (data not shown). The addition of neutralizing antibodies to IL–12 or IFN-γ to cultures of NP-reactive B10.S LN cells (Fig. 5 B) or MBP-reactive SJL LN cells (Fig. 5 C) inhibited the ability of these cells to upregulate IL-12Rβ2 subunit mRNA in response to antigenic stimulation. Thus, the complementary roles of endogenous IL-12 and IFN-γ for optimal expression of IL-12Rβ2 subunit seen in the response of C57BL/6 mice to OVA can be extended to a second foreign antigen (NP) and an autoantigen (MBP).


Regulation of interleukin (IL)-12 receptor beta2 subunit expression by endogenous IL-12: a critical step in the differentiation of pathogenic autoreactive T cells.

Chang JT, Shevach EM, Segal BM - J. Exp. Med. (1999)

Maintenance of IL-12Rβ2  mRNA expression is dependent on the presence of IL-12 and IFN-γ. (A) LN cells from  B10.S mice coimmunized with MBP/NP  were removed on day 12 after immunization  and cultured for 3 d in the presence or absence of MBP with or without IL-12 (2 or  0.2 ng/ml). Total RNA was extracted, and  Northern blot analysis was performed for  IL-12Rβ2 subunit and β-actin. Results are  representative of three experiments each using LN cells pooled from five or more mice. (B) LN cells from B10.S mice coimmunized with MBP/NP were removed on day 12 after immunization and  cultured for 3 d in the presence of NP with or without anti–IL-12 (10 μg /ml) or anti–IFN-γ (10 μg/ml). Total RNA was extracted, and Northern blot  analysis was performed for IL-12Rβ2 subunit and β-actin. Results are representative of three experiments each using LN cells pooled from five or more  mice. (C) LN cells from SJL mice coimmunized with MBP/NP were removed on day 12 after immunization and cultured for 3 d in the presence of MBP  with or without anti–IL-12 (10 μg/ml) or anti–IFN-γ (10 μg/ml). Total RNA was extracted, and Northern blot analysis was performed for IL-12Rβ2  subunit and β-actin. Results are representative of three experiments each using LN cells pooled from five or more mice.
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Figure 5: Maintenance of IL-12Rβ2 mRNA expression is dependent on the presence of IL-12 and IFN-γ. (A) LN cells from B10.S mice coimmunized with MBP/NP were removed on day 12 after immunization and cultured for 3 d in the presence or absence of MBP with or without IL-12 (2 or 0.2 ng/ml). Total RNA was extracted, and Northern blot analysis was performed for IL-12Rβ2 subunit and β-actin. Results are representative of three experiments each using LN cells pooled from five or more mice. (B) LN cells from B10.S mice coimmunized with MBP/NP were removed on day 12 after immunization and cultured for 3 d in the presence of NP with or without anti–IL-12 (10 μg /ml) or anti–IFN-γ (10 μg/ml). Total RNA was extracted, and Northern blot analysis was performed for IL-12Rβ2 subunit and β-actin. Results are representative of three experiments each using LN cells pooled from five or more mice. (C) LN cells from SJL mice coimmunized with MBP/NP were removed on day 12 after immunization and cultured for 3 d in the presence of MBP with or without anti–IL-12 (10 μg/ml) or anti–IFN-γ (10 μg/ml). Total RNA was extracted, and Northern blot analysis was performed for IL-12Rβ2 subunit and β-actin. Results are representative of three experiments each using LN cells pooled from five or more mice.
Mentions: We have previously demonstrated that the addition of pharmacological concentrations of IL-12 to cultures of B10.S MBP-reactive LN cells restored their ability to produce IFN-γ and converted them into encephalitogenic effectors (28). Indeed, the addition of exogenous IL-12 restored the ability of the B10.S MBP-reactive LN cells to upregulate the expression of IL-12Rβ2 subunit mRNA in an antigen-specific, dose-dependent manner (Fig. 5 A). Although the addition of IFN-γ failed to restore the ability of these cells to produce IFN-γ upon secondary stimulation (28), modest induction of IL-12Rβ2 subunit expression was seen (data not shown). The addition of neutralizing antibodies to IL–12 or IFN-γ to cultures of NP-reactive B10.S LN cells (Fig. 5 B) or MBP-reactive SJL LN cells (Fig. 5 C) inhibited the ability of these cells to upregulate IL-12Rβ2 subunit mRNA in response to antigenic stimulation. Thus, the complementary roles of endogenous IL-12 and IFN-γ for optimal expression of IL-12Rβ2 subunit seen in the response of C57BL/6 mice to OVA can be extended to a second foreign antigen (NP) and an autoantigen (MBP).

Bottom Line: Defective expression was not secondary to the production of suppressive cytokines, but to a failure of B10.S MBP-specific T cells to upregulate CD40 ligand expression and to induce the production of IL-12.IL-12Rbeta2 expression as well as encephalitogenicity of these cells could be restored by the addition of IL-12.These results suggest that the development of immunotherapies that target the IL-12Rbeta2 subunit may be useful for the treatment of autoimmune diseases.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.

ABSTRACT
The interleukin (IL)-12 receptor (R)beta2 subunit is the critical molecule involved in maintaining IL-12 responsiveness and controlling T helper cell type 1 lineage commitment. We demonstrate that IL-12 and interferon (IFN)-gamma play separate, but complementary, roles in regulating IL-12Rbeta2 expression on antigen-specific CD4(+) T cells. These results are consistent with our previous observation that IL-12 can promote autoimmune disease through IFN-gamma-independent as well as -dependent pathways. Therefore, we compared the induction of IL-12 by, and the expression of the IL-12Rbeta2 subunit on, myelin basic protein (MBP)-specific T cells from experimental allergic encephalomyelitis (EAE)-susceptible SJL (H-2(s)) mice and from EAE- resistant B10.S mice (H-2(s)). B10.S mice had an antigen-specific defect in their capacity to upregulate the IL-12Rbeta2 subunit. Defective expression was not secondary to the production of suppressive cytokines, but to a failure of B10.S MBP-specific T cells to upregulate CD40 ligand expression and to induce the production of IL-12. IL-12Rbeta2 expression as well as encephalitogenicity of these cells could be restored by the addition of IL-12. These results suggest that the development of immunotherapies that target the IL-12Rbeta2 subunit may be useful for the treatment of autoimmune diseases.

Show MeSH
Related in: MedlinePlus