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Critical role of the programmed death-1 (PD-1) pathway in regulation of experimental autoimmune encephalomyelitis.

Salama AD, Chitnis T, Imitola J, Ansari MJ, Akiba H, Tushima F, Azuma M, Yagita H, Sayegh MH, Khoury SJ - J. Exp. Med. (2003)

Bottom Line: Worsening of disease after PD-1 blockade was associated with a heightened autoimmune response to MOG, manifested by increased frequency of interferon gamma-producing T cells, increased delayed-type hypersensitivity responses, and higher serum levels of anti-MOG antibody.In vivo blockade of PD-1 resulted in increased antigen-specific T cell expansion, activation, and cytokine production.Interestingly, PD-L2 but not PD-L1 blockade in WT animals also resulted in disease augmentation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunogenetics and Transplantation, Children's Hospital, Harvard Medical School, Boston, MA 02115, USA.

ABSTRACT
Experimental autoimmune encephalomyelitis (EAE) is mediated by autoantigen-specific T cells dependent on critical costimulatory signals for their full activation and regulation. We report that the programmed death-1 (PD-1) costimulatory pathway plays a critical role in regulating peripheral tolerance in murine EAE and appears to be a major contributor to the resistance of disease induction in CD28-deficient mice. After immunization with myelin oligodendrocyte glycoprotein (MOG) there was a progressive increase in expression of PD-1 and its ligand PD-L1 but not PD-L2 within the central nervous system (CNS) of mice with EAE, peaking after 3 wk. In both wild-type (WT) and CD28-deficient mice, PD-1 blockade resulted in accelerated and more severe disease with increased CNS lymphocyte infiltration. Worsening of disease after PD-1 blockade was associated with a heightened autoimmune response to MOG, manifested by increased frequency of interferon gamma-producing T cells, increased delayed-type hypersensitivity responses, and higher serum levels of anti-MOG antibody. In vivo blockade of PD-1 resulted in increased antigen-specific T cell expansion, activation, and cytokine production. Interestingly, PD-L2 but not PD-L1 blockade in WT animals also resulted in disease augmentation. Our data are the first demonstration that the PD-1 pathway plays a critical role in regulating EAE.

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Effect of PD-1 blockade on antigen-specific cells in vivo. DO11.10 TCR Tg cells (3 × 106 cells per mouse) were transferred into BALB/c mice. 4 d after priming with OVA peptide, the draining lymph nodes were collected and the number of OVA-specific (KJ1-26+) CD4+ T cells was measured. (a) Flow cytometry plot of CD4+ KJ1-26+ cells from control IgG–treated animals, demonstrating that 1.54% of lymph node cells were TCR transgenic CD4+ T cells. (b) By comparison, in the anti–PD1–treated animal 2.5% of lymph node cells were TCR transgenic. (c) Calculating the absolute numbers of TCR transgenic cells demonstrates a significant increase in KJ1-26+ CD4+ T cells in the anti–PD-1–treated animals compared with controls (P < 0.0001 by two-tailed Mann-Whitney U test).
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fig5: Effect of PD-1 blockade on antigen-specific cells in vivo. DO11.10 TCR Tg cells (3 × 106 cells per mouse) were transferred into BALB/c mice. 4 d after priming with OVA peptide, the draining lymph nodes were collected and the number of OVA-specific (KJ1-26+) CD4+ T cells was measured. (a) Flow cytometry plot of CD4+ KJ1-26+ cells from control IgG–treated animals, demonstrating that 1.54% of lymph node cells were TCR transgenic CD4+ T cells. (b) By comparison, in the anti–PD1–treated animal 2.5% of lymph node cells were TCR transgenic. (c) Calculating the absolute numbers of TCR transgenic cells demonstrates a significant increase in KJ1-26+ CD4+ T cells in the anti–PD-1–treated animals compared with controls (P < 0.0001 by two-tailed Mann-Whitney U test).

Mentions: Because ELISPOT frequencies of MOG-reactive IFN-γ–producing T cells were increased after PD-1 blockade in normal and CD28-deficient mice, we decided to examine the effect of PD-1 blockade on T cell expansion and production of Th1/Th2 cytokines using another antigen-specific system. We used the adoptive transfer of OVA-specific T cells from DO11.10 TCR transgenic mice (6, 19, 20). Naive BALB/c mice were adoptively transferred with DO11.10 splenocytes and their responses after immunization with OVA peptide and treatment with anti–PD-1 mAb or control IgG were examined. The transferred DO11.10 T cells were identified by using an anti-TCR clonotypic mAb KJ1-26 (Fig. 5 , a and b). Control nonimmunized mice demonstrated that the CD4+ KJ1-26+ DO11.10 T cells made up <0.7% of the lymph node cells (corresponding to a total of 1.6 × 103 CD4+ KJ1-26+ cells; not depicted). However, after immunization with OVA peptide, there was a significant expansion of antigen-specific T cells in the draining lymph nodes. In the control IgG–treated animals this reached 5.7 ± 0.6 × 106 cells, whereas in the anti–PD-1–treated animals this reached 91.8 ± 20.9 × 106, reflecting a marked (16-fold) expansion of antigen-specific T cells (Fig. 5 c; P < 0.0001). Furthermore, a greater percentage of cells expressed the activation marker CD25 (94.3 ± 2.2% compared with 76.8 ± 8.9% in anti–PD-1–treated and control mice, respectively). Finally, PD-1 blockade resulted in a greater number of cells producing both Th1 (IFN-γ: 16.7 ± 0.8 × 105 vs. 5.7 ± 0.13 × 105, anti–PD-1 and control, respectively) and Th2 (IL-4: 46.49 ± 7.08 × 105 vs. 4.98 ± 1.46 × 105; IL-5: 32.37 ± 17.9 × 105 vs. 0; and IL-10: 6.25 ± 3.4 × 105 vs. 0) cytokines (not depicted).


Critical role of the programmed death-1 (PD-1) pathway in regulation of experimental autoimmune encephalomyelitis.

Salama AD, Chitnis T, Imitola J, Ansari MJ, Akiba H, Tushima F, Azuma M, Yagita H, Sayegh MH, Khoury SJ - J. Exp. Med. (2003)

Effect of PD-1 blockade on antigen-specific cells in vivo. DO11.10 TCR Tg cells (3 × 106 cells per mouse) were transferred into BALB/c mice. 4 d after priming with OVA peptide, the draining lymph nodes were collected and the number of OVA-specific (KJ1-26+) CD4+ T cells was measured. (a) Flow cytometry plot of CD4+ KJ1-26+ cells from control IgG–treated animals, demonstrating that 1.54% of lymph node cells were TCR transgenic CD4+ T cells. (b) By comparison, in the anti–PD1–treated animal 2.5% of lymph node cells were TCR transgenic. (c) Calculating the absolute numbers of TCR transgenic cells demonstrates a significant increase in KJ1-26+ CD4+ T cells in the anti–PD-1–treated animals compared with controls (P < 0.0001 by two-tailed Mann-Whitney U test).
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Related In: Results  -  Collection

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fig5: Effect of PD-1 blockade on antigen-specific cells in vivo. DO11.10 TCR Tg cells (3 × 106 cells per mouse) were transferred into BALB/c mice. 4 d after priming with OVA peptide, the draining lymph nodes were collected and the number of OVA-specific (KJ1-26+) CD4+ T cells was measured. (a) Flow cytometry plot of CD4+ KJ1-26+ cells from control IgG–treated animals, demonstrating that 1.54% of lymph node cells were TCR transgenic CD4+ T cells. (b) By comparison, in the anti–PD1–treated animal 2.5% of lymph node cells were TCR transgenic. (c) Calculating the absolute numbers of TCR transgenic cells demonstrates a significant increase in KJ1-26+ CD4+ T cells in the anti–PD-1–treated animals compared with controls (P < 0.0001 by two-tailed Mann-Whitney U test).
Mentions: Because ELISPOT frequencies of MOG-reactive IFN-γ–producing T cells were increased after PD-1 blockade in normal and CD28-deficient mice, we decided to examine the effect of PD-1 blockade on T cell expansion and production of Th1/Th2 cytokines using another antigen-specific system. We used the adoptive transfer of OVA-specific T cells from DO11.10 TCR transgenic mice (6, 19, 20). Naive BALB/c mice were adoptively transferred with DO11.10 splenocytes and their responses after immunization with OVA peptide and treatment with anti–PD-1 mAb or control IgG were examined. The transferred DO11.10 T cells were identified by using an anti-TCR clonotypic mAb KJ1-26 (Fig. 5 , a and b). Control nonimmunized mice demonstrated that the CD4+ KJ1-26+ DO11.10 T cells made up <0.7% of the lymph node cells (corresponding to a total of 1.6 × 103 CD4+ KJ1-26+ cells; not depicted). However, after immunization with OVA peptide, there was a significant expansion of antigen-specific T cells in the draining lymph nodes. In the control IgG–treated animals this reached 5.7 ± 0.6 × 106 cells, whereas in the anti–PD-1–treated animals this reached 91.8 ± 20.9 × 106, reflecting a marked (16-fold) expansion of antigen-specific T cells (Fig. 5 c; P < 0.0001). Furthermore, a greater percentage of cells expressed the activation marker CD25 (94.3 ± 2.2% compared with 76.8 ± 8.9% in anti–PD-1–treated and control mice, respectively). Finally, PD-1 blockade resulted in a greater number of cells producing both Th1 (IFN-γ: 16.7 ± 0.8 × 105 vs. 5.7 ± 0.13 × 105, anti–PD-1 and control, respectively) and Th2 (IL-4: 46.49 ± 7.08 × 105 vs. 4.98 ± 1.46 × 105; IL-5: 32.37 ± 17.9 × 105 vs. 0; and IL-10: 6.25 ± 3.4 × 105 vs. 0) cytokines (not depicted).

Bottom Line: Worsening of disease after PD-1 blockade was associated with a heightened autoimmune response to MOG, manifested by increased frequency of interferon gamma-producing T cells, increased delayed-type hypersensitivity responses, and higher serum levels of anti-MOG antibody.In vivo blockade of PD-1 resulted in increased antigen-specific T cell expansion, activation, and cytokine production.Interestingly, PD-L2 but not PD-L1 blockade in WT animals also resulted in disease augmentation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunogenetics and Transplantation, Children's Hospital, Harvard Medical School, Boston, MA 02115, USA.

ABSTRACT
Experimental autoimmune encephalomyelitis (EAE) is mediated by autoantigen-specific T cells dependent on critical costimulatory signals for their full activation and regulation. We report that the programmed death-1 (PD-1) costimulatory pathway plays a critical role in regulating peripheral tolerance in murine EAE and appears to be a major contributor to the resistance of disease induction in CD28-deficient mice. After immunization with myelin oligodendrocyte glycoprotein (MOG) there was a progressive increase in expression of PD-1 and its ligand PD-L1 but not PD-L2 within the central nervous system (CNS) of mice with EAE, peaking after 3 wk. In both wild-type (WT) and CD28-deficient mice, PD-1 blockade resulted in accelerated and more severe disease with increased CNS lymphocyte infiltration. Worsening of disease after PD-1 blockade was associated with a heightened autoimmune response to MOG, manifested by increased frequency of interferon gamma-producing T cells, increased delayed-type hypersensitivity responses, and higher serum levels of anti-MOG antibody. In vivo blockade of PD-1 resulted in increased antigen-specific T cell expansion, activation, and cytokine production. Interestingly, PD-L2 but not PD-L1 blockade in WT animals also resulted in disease augmentation. Our data are the first demonstration that the PD-1 pathway plays a critical role in regulating EAE.

Show MeSH
Related in: MedlinePlus