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Pharmacological prion protein silencing accelerates central nervous system autoimmune disease via T cell receptor signalling.

Hu W, Nessler S, Hemmer B, Eagar TN, Kane LP, Leliveld SR, Müller-Schiffmann A, Gocke AR, Lovett-Racke A, Ben LH, Hussain RZ, Breil A, Elliott JL, Puttaparthi K, Cravens PD, Singh MP, Petsch B, Stitz L, Racke MK, Korth C, Stüve O - Brain (2010)

Bottom Line: Disruption of cellular prion protein signalling augmented antigen-specific activation and proliferation, and enhanced T cell receptor signalling, resulting in zeta-chain-associated protein-70 phosphorylation and nuclear factor of activated T cells/activator protein 1 transcriptional activity.Cellular prion protein silencing with small interfering ribonucleic acid also resulted in the worsening of actively induced and adoptively transferred experimental autoimmune encephalomyelitis.Thus, central nervous system autoimmune disease was modulated at all stages of disease: the generation of the T cell effector response, the elicitation of T effector function and the perpetuation of cellular immune responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, University of Texas Southwestern Medical Center at Dallas, TX, USA.

ABSTRACT
The primary biological function of the endogenous cellular prion protein has remained unclear. We investigated its biological function in the generation of cellular immune responses using cellular prion protein gene-specific small interfering ribonucleic acid in vivo and in vitro. Our results were confirmed by blocking cellular prion protein with monovalent antibodies and by using cellular prion protein-deficient and -transgenic mice. In vivo prion protein gene-small interfering ribonucleic acid treatment effects were of limited duration, restricted to secondary lymphoid organs and resulted in a 70% reduction of cellular prion protein expression in leukocytes. Disruption of cellular prion protein signalling augmented antigen-specific activation and proliferation, and enhanced T cell receptor signalling, resulting in zeta-chain-associated protein-70 phosphorylation and nuclear factor of activated T cells/activator protein 1 transcriptional activity. In vivo prion protein gene-small interfering ribonucleic acid treatment promoted T cell differentiation towards pro-inflammatory phenotypes and increased survival of antigen-specific T cells. Cellular prion protein silencing with small interfering ribonucleic acid also resulted in the worsening of actively induced and adoptively transferred experimental autoimmune encephalomyelitis. Finally, treatment of myelin basic protein(1-11) T cell receptor transgenic mice with prion protein gene-small interfering ribonucleic acid resulted in spontaneous experimental autoimmune encephalomyelitis. Thus, central nervous system autoimmune disease was modulated at all stages of disease: the generation of the T cell effector response, the elicitation of T effector function and the perpetuation of cellular immune responses. Our findings indicate that cellular prion protein regulates T cell receptor-mediated T cell activation, differentiation and survival. Defects in autoimmunity are restricted to the immune system and not the central nervous system. Our data identify cellular prion protein as a regulator of cellular immunological homoeostasis and suggest cellular prion protein as a novel potential target for therapeutic immunomodulation.

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Prnp-siRNA treated and PrP-deficient (PrP−/−) mice develop more severe EAE whereas mice overexpressing PrPC are protected. (A and B) In vivo silencing of PrPc worsens the clinical course of actively induced EAE in SJL/J mice. (A) In an EAE ‘prevention’ experiment, SJL/L mice were injected intravenously with a single dose of Prnp-siRNA or nonsense (NS)-siRNA at the time of immunization with PLPp139−151, as indicated by a grey arrow. Prnp-siRNA treatment resulted in clinically more severe disease. (B) In an EAE ‘treatment’ experiment, Prnp-siRNA or nonsense-siRNA was injected at the time of clinical onset of EAE (grey arrow). Mice treated with Prnp-siRNA had a significantly worse initial clinical exacerbation. (C) Male and (D) female PrP−/− mice had earlier disease onset and significantly higher disease scores than male wild-type (WT) mice. (E) Tga20 mice had a delayed disease onset, and developed only mild EAE compared to C57BL/6 and Sv129 wild-type mice.
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Figure 2: Prnp-siRNA treated and PrP-deficient (PrP−/−) mice develop more severe EAE whereas mice overexpressing PrPC are protected. (A and B) In vivo silencing of PrPc worsens the clinical course of actively induced EAE in SJL/J mice. (A) In an EAE ‘prevention’ experiment, SJL/L mice were injected intravenously with a single dose of Prnp-siRNA or nonsense (NS)-siRNA at the time of immunization with PLPp139−151, as indicated by a grey arrow. Prnp-siRNA treatment resulted in clinically more severe disease. (B) In an EAE ‘treatment’ experiment, Prnp-siRNA or nonsense-siRNA was injected at the time of clinical onset of EAE (grey arrow). Mice treated with Prnp-siRNA had a significantly worse initial clinical exacerbation. (C) Male and (D) female PrP−/− mice had earlier disease onset and significantly higher disease scores than male wild-type (WT) mice. (E) Tga20 mice had a delayed disease onset, and developed only mild EAE compared to C57BL/6 and Sv129 wild-type mice.

Mentions: To test the role of PrPc in CNS autoimmune disease, EAE was induced by active immunization of SJL/J mice with PLPp139−151. A single dose of 50 μg Prnp-siRNA or nonsense-siRNA was injected i.v. via the tail vein in 100 μl PBS according to published methods (Lovett-Racke et al., 2004). Experimental animals were monitored for 40 days. In a set of ‘prevention’ experiments, treatment with Prnp-siRNA resulted in significantly worse clinical EAE within the first 10 days after disease onset than treatment with nonsense-siRNA or no treatment (Fig. 2A, Supplementary Table S1A). In a second set of ‘treatment’ experiments, mice treated with Prnp-siRNA had a significantly worse initial clinical exacerbation than control mice and continued to do significantly worse throughout the observation period (Fig. 2B, Supplementary Table S1B)Figure 2


Pharmacological prion protein silencing accelerates central nervous system autoimmune disease via T cell receptor signalling.

Hu W, Nessler S, Hemmer B, Eagar TN, Kane LP, Leliveld SR, Müller-Schiffmann A, Gocke AR, Lovett-Racke A, Ben LH, Hussain RZ, Breil A, Elliott JL, Puttaparthi K, Cravens PD, Singh MP, Petsch B, Stitz L, Racke MK, Korth C, Stüve O - Brain (2010)

Prnp-siRNA treated and PrP-deficient (PrP−/−) mice develop more severe EAE whereas mice overexpressing PrPC are protected. (A and B) In vivo silencing of PrPc worsens the clinical course of actively induced EAE in SJL/J mice. (A) In an EAE ‘prevention’ experiment, SJL/L mice were injected intravenously with a single dose of Prnp-siRNA or nonsense (NS)-siRNA at the time of immunization with PLPp139−151, as indicated by a grey arrow. Prnp-siRNA treatment resulted in clinically more severe disease. (B) In an EAE ‘treatment’ experiment, Prnp-siRNA or nonsense-siRNA was injected at the time of clinical onset of EAE (grey arrow). Mice treated with Prnp-siRNA had a significantly worse initial clinical exacerbation. (C) Male and (D) female PrP−/− mice had earlier disease onset and significantly higher disease scores than male wild-type (WT) mice. (E) Tga20 mice had a delayed disease onset, and developed only mild EAE compared to C57BL/6 and Sv129 wild-type mice.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
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Figure 2: Prnp-siRNA treated and PrP-deficient (PrP−/−) mice develop more severe EAE whereas mice overexpressing PrPC are protected. (A and B) In vivo silencing of PrPc worsens the clinical course of actively induced EAE in SJL/J mice. (A) In an EAE ‘prevention’ experiment, SJL/L mice were injected intravenously with a single dose of Prnp-siRNA or nonsense (NS)-siRNA at the time of immunization with PLPp139−151, as indicated by a grey arrow. Prnp-siRNA treatment resulted in clinically more severe disease. (B) In an EAE ‘treatment’ experiment, Prnp-siRNA or nonsense-siRNA was injected at the time of clinical onset of EAE (grey arrow). Mice treated with Prnp-siRNA had a significantly worse initial clinical exacerbation. (C) Male and (D) female PrP−/− mice had earlier disease onset and significantly higher disease scores than male wild-type (WT) mice. (E) Tga20 mice had a delayed disease onset, and developed only mild EAE compared to C57BL/6 and Sv129 wild-type mice.
Mentions: To test the role of PrPc in CNS autoimmune disease, EAE was induced by active immunization of SJL/J mice with PLPp139−151. A single dose of 50 μg Prnp-siRNA or nonsense-siRNA was injected i.v. via the tail vein in 100 μl PBS according to published methods (Lovett-Racke et al., 2004). Experimental animals were monitored for 40 days. In a set of ‘prevention’ experiments, treatment with Prnp-siRNA resulted in significantly worse clinical EAE within the first 10 days after disease onset than treatment with nonsense-siRNA or no treatment (Fig. 2A, Supplementary Table S1A). In a second set of ‘treatment’ experiments, mice treated with Prnp-siRNA had a significantly worse initial clinical exacerbation than control mice and continued to do significantly worse throughout the observation period (Fig. 2B, Supplementary Table S1B)Figure 2

Bottom Line: Disruption of cellular prion protein signalling augmented antigen-specific activation and proliferation, and enhanced T cell receptor signalling, resulting in zeta-chain-associated protein-70 phosphorylation and nuclear factor of activated T cells/activator protein 1 transcriptional activity.Cellular prion protein silencing with small interfering ribonucleic acid also resulted in the worsening of actively induced and adoptively transferred experimental autoimmune encephalomyelitis.Thus, central nervous system autoimmune disease was modulated at all stages of disease: the generation of the T cell effector response, the elicitation of T effector function and the perpetuation of cellular immune responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, University of Texas Southwestern Medical Center at Dallas, TX, USA.

ABSTRACT
The primary biological function of the endogenous cellular prion protein has remained unclear. We investigated its biological function in the generation of cellular immune responses using cellular prion protein gene-specific small interfering ribonucleic acid in vivo and in vitro. Our results were confirmed by blocking cellular prion protein with monovalent antibodies and by using cellular prion protein-deficient and -transgenic mice. In vivo prion protein gene-small interfering ribonucleic acid treatment effects were of limited duration, restricted to secondary lymphoid organs and resulted in a 70% reduction of cellular prion protein expression in leukocytes. Disruption of cellular prion protein signalling augmented antigen-specific activation and proliferation, and enhanced T cell receptor signalling, resulting in zeta-chain-associated protein-70 phosphorylation and nuclear factor of activated T cells/activator protein 1 transcriptional activity. In vivo prion protein gene-small interfering ribonucleic acid treatment promoted T cell differentiation towards pro-inflammatory phenotypes and increased survival of antigen-specific T cells. Cellular prion protein silencing with small interfering ribonucleic acid also resulted in the worsening of actively induced and adoptively transferred experimental autoimmune encephalomyelitis. Finally, treatment of myelin basic protein(1-11) T cell receptor transgenic mice with prion protein gene-small interfering ribonucleic acid resulted in spontaneous experimental autoimmune encephalomyelitis. Thus, central nervous system autoimmune disease was modulated at all stages of disease: the generation of the T cell effector response, the elicitation of T effector function and the perpetuation of cellular immune responses. Our findings indicate that cellular prion protein regulates T cell receptor-mediated T cell activation, differentiation and survival. Defects in autoimmunity are restricted to the immune system and not the central nervous system. Our data identify cellular prion protein as a regulator of cellular immunological homoeostasis and suggest cellular prion protein as a novel potential target for therapeutic immunomodulation.

Show MeSH
Related in: MedlinePlus