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Interferon-β suppresses murine Th1 cell function in the absence of antigen-presenting cells.

Boivin N, Baillargeon J, Doss PM, Roy AP, Rangachari M - PLoS ONE (2015)

Bottom Line: CD4+ T cells express the type I interferon receptor, and IFN-β can suppress Th1 cell proliferation under APC-free stimulation conditions.Further, IFN-β treatment of Th1 cells upregulates phosphorylation of Stat1, and downregulates phosphorylation of Stat4.Our data indicate that IFN-γ-producing Th1 cells are directly responsive to IFN-β and point to a novel mechanism of IFN-β-mediated T cell suppression that is independent of APC-derived signals.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroscience, Centre de recherche du CHU de Québec-Université Laval, Québec QC, Canada G1V 4G2.

ABSTRACT
Interferon (IFN)-β is a front-line therapy for the treatment of the relapsing-remitting form of multiple sclerosis. However, its immunosuppressive mechanism of function remains incompletely understood. While it has been proposed that IFN-β suppresses the function of inflammatory myelin antigen-reactive T cells by promoting the release of immunomodulatory cytokines such as IL-27 from antigen-presenting cells (APCs), its direct effects on inflammatory CD4+ Th1 cells are less clear. Here, we establish that IFN-β inhibits mouse IFN-γ+ Th1 cell function in the absence of APCs. CD4+ T cells express the type I interferon receptor, and IFN-β can suppress Th1 cell proliferation under APC-free stimulation conditions. IFN-β-treated myelin antigen-specific Th1 cells are impaired in their ability to induce severe experimental autoimmune encephalomyelitis (EAE) upon transfer to lymphocyte-deficient Rag1-/- mice. Polarized Th1 cells downregulate IFN-γ and IL-2, and upregulate the negative regulatory receptor Tim-3, when treated with IFN-β in the absence of APCs. Further, IFN-β treatment of Th1 cells upregulates phosphorylation of Stat1, and downregulates phosphorylation of Stat4. Our data indicate that IFN-γ-producing Th1 cells are directly responsive to IFN-β and point to a novel mechanism of IFN-β-mediated T cell suppression that is independent of APC-derived signals.

No MeSH data available.


Related in: MedlinePlus

IFN-β suppresses Th1 cell proliferation.A. WT CD4+CD62Lhi T cells were labeled with CFSE and stimulated for 5 days with either soluble anti-CD3 plus irradiated splenocytes (APCs), or with plate-bound anti-CD3+anti-CD28, under Th1 or Th17 conditions, with the indicated concentrations of IFN-β. CFSE dilution was assessed by flow cytometry. Data representative of three experiments. B. WT or IFNAR1-/- CD4+CD62Lhi T cells were labeled with CFSE and stimulated with plate-bound anti-CD3+anti-CD28 under Th1 conditions with the indicated concentrations of IFN-β for 5 days. CFSE dilution was assessed by flow cytometry. Data representative of two experiments. Gates represent the percentage of cells that underwent at least one division.
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pone.0124802.g002: IFN-β suppresses Th1 cell proliferation.A. WT CD4+CD62Lhi T cells were labeled with CFSE and stimulated for 5 days with either soluble anti-CD3 plus irradiated splenocytes (APCs), or with plate-bound anti-CD3+anti-CD28, under Th1 or Th17 conditions, with the indicated concentrations of IFN-β. CFSE dilution was assessed by flow cytometry. Data representative of three experiments. B. WT or IFNAR1-/- CD4+CD62Lhi T cells were labeled with CFSE and stimulated with plate-bound anti-CD3+anti-CD28 under Th1 conditions with the indicated concentrations of IFN-β for 5 days. CFSE dilution was assessed by flow cytometry. Data representative of two experiments. Gates represent the percentage of cells that underwent at least one division.

Mentions: Having determined that Th1 cells express IFN-α/β-R, we next wanted to determine whether they could respond to treatment with IFN-β. Previous studies had analyzed the effects of IFN-β function on Th1 cells while using B cells [19], fibroblasts [20,22], irradiated splenocytes [14,31], purified dendritic cells [32] or whole blood leukocytes [33] as APCs. Here, we wished to examine whether IFN-β could modulate Th1 cell function in the absence of APCs. Thus, we stimulated naïve CD62Lhi CD4+ T cells with immobilized anti-CD3 and anti-CD28 monoclonal antibodies (plate-bound stimulation). This protocol mimics physiologic activation of T cells via concomitant stimulation through both their T cell receptor and the costimulatory receptor CD28. Using this plate-bound approach allows us to isolate the effects of IFN-β on T cells in the absence of APC-derived cues. To generate Th1 cells, we stimulated naïve CD62Lhi CD4+ T cells in vitro in the presence of the Th1 driving cytokine IL-12. In addition, we used a blocking antibody to IL-4 in order to suppress Th2 cell differentiation. As expected, IFN-β suppressed the proliferation of Th1 cells cultured with APCs, as measured by CFSE dilution. Interestingly, IFN-β also suppressed the proliferation of Th1 cells stimulated by anti-CD3+anti-CD28, indicating that its antiproliferative effects do not require APC-derived signals (Fig 2A). This effect was dependent on the expression of IFN-α/β-R, as IFN-β had little effect on the proliferation of IFNAR1-/- Th1 cells stimulated under APC-free conditions (Fig 2B). As we found that IFN-α/β-R is more highly expressed on Th1 cells than Th17 cells (Fig 1B), we wanted to ascertain whether IFN-β differentially regulates the proliferation of Th1 versus Th17 cells. Thus, we generated Th17 cells by stimulating CD62LhiCD4+ T cells with anti-CD3 and anti-CD28 in the presence of TGF-β and IL-6 [34], and in the presence (10 U mL-1, 100 U mL-1) or absence of IFN-β. We found that while IFN-β could suppress Th17 cell proliferation, it only did so at a dose of 100 U mL-1. By contrast, as little as 10 U ml-1 IFN-β inhibited Th1 cell proliferation (Fig 2A). Taken together, these data indicate that IFN-β can suppress the proliferation of Th1 and Th17 cells in the absence of APCs and that this effect is more pronounced in Th1 cells.


Interferon-β suppresses murine Th1 cell function in the absence of antigen-presenting cells.

Boivin N, Baillargeon J, Doss PM, Roy AP, Rangachari M - PLoS ONE (2015)

IFN-β suppresses Th1 cell proliferation.A. WT CD4+CD62Lhi T cells were labeled with CFSE and stimulated for 5 days with either soluble anti-CD3 plus irradiated splenocytes (APCs), or with plate-bound anti-CD3+anti-CD28, under Th1 or Th17 conditions, with the indicated concentrations of IFN-β. CFSE dilution was assessed by flow cytometry. Data representative of three experiments. B. WT or IFNAR1-/- CD4+CD62Lhi T cells were labeled with CFSE and stimulated with plate-bound anti-CD3+anti-CD28 under Th1 conditions with the indicated concentrations of IFN-β for 5 days. CFSE dilution was assessed by flow cytometry. Data representative of two experiments. Gates represent the percentage of cells that underwent at least one division.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4401451&req=5

pone.0124802.g002: IFN-β suppresses Th1 cell proliferation.A. WT CD4+CD62Lhi T cells were labeled with CFSE and stimulated for 5 days with either soluble anti-CD3 plus irradiated splenocytes (APCs), or with plate-bound anti-CD3+anti-CD28, under Th1 or Th17 conditions, with the indicated concentrations of IFN-β. CFSE dilution was assessed by flow cytometry. Data representative of three experiments. B. WT or IFNAR1-/- CD4+CD62Lhi T cells were labeled with CFSE and stimulated with plate-bound anti-CD3+anti-CD28 under Th1 conditions with the indicated concentrations of IFN-β for 5 days. CFSE dilution was assessed by flow cytometry. Data representative of two experiments. Gates represent the percentage of cells that underwent at least one division.
Mentions: Having determined that Th1 cells express IFN-α/β-R, we next wanted to determine whether they could respond to treatment with IFN-β. Previous studies had analyzed the effects of IFN-β function on Th1 cells while using B cells [19], fibroblasts [20,22], irradiated splenocytes [14,31], purified dendritic cells [32] or whole blood leukocytes [33] as APCs. Here, we wished to examine whether IFN-β could modulate Th1 cell function in the absence of APCs. Thus, we stimulated naïve CD62Lhi CD4+ T cells with immobilized anti-CD3 and anti-CD28 monoclonal antibodies (plate-bound stimulation). This protocol mimics physiologic activation of T cells via concomitant stimulation through both their T cell receptor and the costimulatory receptor CD28. Using this plate-bound approach allows us to isolate the effects of IFN-β on T cells in the absence of APC-derived cues. To generate Th1 cells, we stimulated naïve CD62Lhi CD4+ T cells in vitro in the presence of the Th1 driving cytokine IL-12. In addition, we used a blocking antibody to IL-4 in order to suppress Th2 cell differentiation. As expected, IFN-β suppressed the proliferation of Th1 cells cultured with APCs, as measured by CFSE dilution. Interestingly, IFN-β also suppressed the proliferation of Th1 cells stimulated by anti-CD3+anti-CD28, indicating that its antiproliferative effects do not require APC-derived signals (Fig 2A). This effect was dependent on the expression of IFN-α/β-R, as IFN-β had little effect on the proliferation of IFNAR1-/- Th1 cells stimulated under APC-free conditions (Fig 2B). As we found that IFN-α/β-R is more highly expressed on Th1 cells than Th17 cells (Fig 1B), we wanted to ascertain whether IFN-β differentially regulates the proliferation of Th1 versus Th17 cells. Thus, we generated Th17 cells by stimulating CD62LhiCD4+ T cells with anti-CD3 and anti-CD28 in the presence of TGF-β and IL-6 [34], and in the presence (10 U mL-1, 100 U mL-1) or absence of IFN-β. We found that while IFN-β could suppress Th17 cell proliferation, it only did so at a dose of 100 U mL-1. By contrast, as little as 10 U ml-1 IFN-β inhibited Th1 cell proliferation (Fig 2A). Taken together, these data indicate that IFN-β can suppress the proliferation of Th1 and Th17 cells in the absence of APCs and that this effect is more pronounced in Th1 cells.

Bottom Line: CD4+ T cells express the type I interferon receptor, and IFN-β can suppress Th1 cell proliferation under APC-free stimulation conditions.Further, IFN-β treatment of Th1 cells upregulates phosphorylation of Stat1, and downregulates phosphorylation of Stat4.Our data indicate that IFN-γ-producing Th1 cells are directly responsive to IFN-β and point to a novel mechanism of IFN-β-mediated T cell suppression that is independent of APC-derived signals.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroscience, Centre de recherche du CHU de Québec-Université Laval, Québec QC, Canada G1V 4G2.

ABSTRACT
Interferon (IFN)-β is a front-line therapy for the treatment of the relapsing-remitting form of multiple sclerosis. However, its immunosuppressive mechanism of function remains incompletely understood. While it has been proposed that IFN-β suppresses the function of inflammatory myelin antigen-reactive T cells by promoting the release of immunomodulatory cytokines such as IL-27 from antigen-presenting cells (APCs), its direct effects on inflammatory CD4+ Th1 cells are less clear. Here, we establish that IFN-β inhibits mouse IFN-γ+ Th1 cell function in the absence of APCs. CD4+ T cells express the type I interferon receptor, and IFN-β can suppress Th1 cell proliferation under APC-free stimulation conditions. IFN-β-treated myelin antigen-specific Th1 cells are impaired in their ability to induce severe experimental autoimmune encephalomyelitis (EAE) upon transfer to lymphocyte-deficient Rag1-/- mice. Polarized Th1 cells downregulate IFN-γ and IL-2, and upregulate the negative regulatory receptor Tim-3, when treated with IFN-β in the absence of APCs. Further, IFN-β treatment of Th1 cells upregulates phosphorylation of Stat1, and downregulates phosphorylation of Stat4. Our data indicate that IFN-γ-producing Th1 cells are directly responsive to IFN-β and point to a novel mechanism of IFN-β-mediated T cell suppression that is independent of APC-derived signals.

No MeSH data available.


Related in: MedlinePlus