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Toll-Like Receptor 7 Preconditioning Induces Robust Neuroprotection Against Stroke by a Novel Type I Interferon-Mediated Mechanism
Philberta Y. Leung, Susan L. Stevens, Amy E.B. Packard, Nikola S. Lessov, Tao Yang, Valerie K. Conrad, Noortje N.A.M. van den Dungen, Roger P. Simon and Mary P. Stenzel-Poore

Stroke. 2012;43:1383-1389, originally published April 23, 2012
https://doi.org/10.1161/STROKEAHA.111.641522

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Toll-Like Receptor 7 Preconditioning Induces Robust Neuroprotection Against Stroke by a Novel Type I Interferon-Mediated Mechanism
Philberta Y. Leung, Susan L. Stevens, Amy E.B. Packard, Nikola S. Lessov, Tao Yang, Valerie K. Conrad, Noortje N.A.M. van den Dungen, Roger P. Simon, Mary P. Stenzel-Poore

Stroke May 2012, 43 (5) 1383-1389; DOI: https://doi.org/10.1161/STROKEAHA.111.641522

Figure 1.
Gardiquimod (GDQ) preconditioning reduces ischemic injury. A, C57Bl/6 mice were preconditio...
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Gardiquimod (GDQ) preconditioning reduces ischemic injury. A, C57Bl/6 mice were preconditioned with escalating doses of GDQ (N=8; 10, 20, or 40 μg per mouse, subcutaneous) or saline (N=5) 72 hours before 60 minutes of middle cerebral artery occulusion (MCAO). Infarct size was determined 24 hours after MCAO. B, C57Bl/6 mice were pretreated with GDQ (N=8; 20 μg/mouse, subcutaneous) or saline (n=6) 72 hours before MCAO. Infarct size was determined 72 hours after MCAO. C, C57Bl/6 mice were pretreated with GDQ (N=5–6; 20 μg/mouse, subcutaneous) or saline (n=6) at various times before MCAO. Infarct size was determined 24 hours after MCAO. Two-way analysis of variance (ANOVA), Bonferroni post hoc, *P<0.01, **P<0.01, ***P<0.001 vs saline controls. D and E, C57BL/6 mice were treated with GDQ (N=5; 40 μg/mouse, subcutaneous) or saline (N=6) 72 hours before MCAO (60 minutes). Mice were then examined using the (D) neurological score (focal and general) and (E) corner test to determine neurological and sensorimotor deficits 24 hours after MCAO. Student t test: *P<0.01, **P<0.01, ***P<0.001 vs saline controls.
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Toll-Like Receptor 7 Preconditioning Induces Robust Neuroprotection Against Stroke by a Novel Type I Interferon-Mediated Mechanism
Philberta Y. Leung, Susan L. Stevens, Amy E.B. Packard, Nikola S. Lessov, Tao Yang, Valerie K. Conrad, Noortje N.A.M. van den Dungen, Roger P. Simon, Mary P. Stenzel-Poore

Stroke May 2012, 43 (5) 1383-1389; DOI: https://doi.org/10.1161/STROKEAHA.111.641522

Figure 2.
Gardiquimod (GDQ)-induced neuroprotection is mediated through Toll-like receptor (TLR) 7. TLR7^+/+...
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Gardiquimod (GDQ)-induced neuroprotection is mediated through Toll-like receptor (TLR) 7. TLR7^+/+ (N=7) or TLR7^−/− (N=5–7) mice were preconditioned with GDQ (40 μg/mouse, subcutaneous) or saline 72 hours before middle cerebral artery occlusion (MCAO; 45 minutes). Infarct size was determined 24 hours after MCAO. Two-way analysis of variance (ANOVA), Bonferroni post hoc, **P<0.01 vs saline control for respective genotype.
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Toll-Like Receptor 7 Preconditioning Induces Robust Neuroprotection Against Stroke by a Novel Type I Interferon-Mediated Mechanism
Philberta Y. Leung, Susan L. Stevens, Amy E.B. Packard, Nikola S. Lessov, Tao Yang, Valerie K. Conrad, Noortje N.A.M. van den Dungen, Roger P. Simon, Mary P. Stenzel-Poore

Stroke May 2012, 43 (5) 1383-1389; DOI: https://doi.org/10.1161/STROKEAHA.111.641522

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Table.
Interferon-Associated Genes in the Brains of Gardiquimod-Preconditioned Animals After Middle Cerebral Artery Occlusion (24 Hours)Show More

Interferon-Associated Genes in the Brains of Gardiquimod-Preconditioned Animals After Middle Cerebral Artery Occlusion (24 Hours)Show Less
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Toll-Like Receptor 7 Preconditioning Induces Robust Neuroprotection Against Stroke by a Novel Type I Interferon-Mediated Mechanism
Philberta Y. Leung, Susan L. Stevens, Amy E.B. Packard, Nikola S. Lessov, Tao Yang, Valerie K. Conrad, Noortje N.A.M. van den Dungen, Roger P. Simon, Mary P. Stenzel-Poore

Stroke May 2012, 43 (5) 1383-1389; DOI: https://doi.org/10.1161/STROKEAHA.111.641522

Figure 3.
Tumor necrosis factor (TNF) is not required for Toll-like receptor (TLR)7-induced neuroprotection. A...
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Tumor necrosis factor (TNF) is not required for Toll-like receptor (TLR)7-induced neuroprotection. A, C57Bl/6 mice were injected with either Gardiquimod (GDQ; 40 μg/mouse, subcutaneous), lipopolysaccharide (LPS; 20 μg/mouse, subcutaneous), or saline. Blood was collected at various time points after injection and serum levels of TNF were determined via enzyme-linked immunosorbent assay (ELISA). N.D.=not detected (N=4 – 6). B, TNF^−/− (N=6 – 8) and TNF^+/+ mice (N=7) were injected with GDQ (40 μg/mouse, subcutaneous) or saline 72 hours before middle cerebral artery occlusion (MCAO; 50 minutes). Infarct size was determined 24 hours after MCAO. Two-way analysis of variance (ANOVA), Bonferroni post hoc: *P<0.05 vs saline controls.
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Toll-Like Receptor 7 Preconditioning Induces Robust Neuroprotection Against Stroke by a Novel Type I Interferon-Mediated Mechanism
Philberta Y. Leung, Susan L. Stevens, Amy E.B. Packard, Nikola S. Lessov, Tao Yang, Valerie K. Conrad, Noortje N.A.M. van den Dungen, Roger P. Simon, Mary P. Stenzel-Poore

Stroke May 2012, 43 (5) 1383-1389; DOI: https://doi.org/10.1161/STROKEAHA.111.641522

Figure 4.
Gardiquimod (GDQ)-preconditioning induces IFNα and requires IRF7 for neuroprotection. A, C5...
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Gardiquimod (GDQ)-preconditioning induces IFNα and requires IRF7 for neuroprotection. A, C57Bl/6 mice were injected with GDQ (40 μg/mouse, subcutaneous), unmethylated cytosine-phosphate-guanine rich oligonucleotide (40 μg/mouse, subcutaneous), lipopolysaccharide (LPS; 20 μg/mouse, subcutaneous), or saline. Blood was collected at indicated times and serum interferon (IFN)α levels were measured (N=4–10). Two-way analysis of variance (ANOVA), Bonferroni post hoc: *P<0.05, ***P<0.001 vs saline controls. B, IRF7^−/− and IRF7^+/+ mice were injected with 40 μg GDQ or saline. Blood was collected at 2 hours and serum IFNα was measured (N=3 – 8). One-way ANOVA, Bonferroni post hoc: *P<0.05 vs saline controls. C, IRF7^+/+ (N=8–9) or IRF7^−/− mice (N=7–9) were preconditioned with GDQ (40 μg/mouse, subcutaneous) or saline 72 hours before MCAO (45 minutes). Infarct size was determined 24 hours after middle cerebral artery occlusion (MCAO). Two-way ANOVA, Bonferroni post hoc: **P<0.01 vs saline controls.
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Toll-Like Receptor 7 Preconditioning Induces Robust Neuroprotection Against Stroke by a Novel Type I Interferon-Mediated Mechanism
Philberta Y. Leung, Susan L. Stevens, Amy E.B. Packard, Nikola S. Lessov, Tao Yang, Valerie K. Conrad, Noortje N.A.M. van den Dungen, Roger P. Simon, Mary P. Stenzel-Poore

Stroke May 2012, 43 (5) 1383-1389; DOI: https://doi.org/10.1161/STROKEAHA.111.641522

Figure 5.
Type I interferon receptor (IFNAR) signaling is required for TLR7-mediated neuroprotection. IFNAR^+/+...
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Type I interferon receptor (IFNAR) signaling is required for TLR7-mediated neuroprotection. IFNAR^+/+ and IFNAR^−/− mice were injected with Gardiquimod (GDQ; N=6 – 8), unmethylated cytosine-phosphate-guanine rich oligonucleotide (N=8–11), lipopolysaccharide (LPS; N=8–11), or saline (N=6 – 9) 72 hours before middle cerebral artery occlusion (MCAO; 45 minutes). Infarct size was determined 24 hours after MCAO. The data are presented as percent damage normalized to saline plus MCAO. Two-way analysis of variance (ANOVA), Bonferroni post hoc: ***P<0.001 vs saline control for respective genotype; #P<0.05 vs IFNAR^+/+ for respective treatment.
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Toll-Like Receptor 7 Preconditioning Induces Robust Neuroprotection Against Stroke by a Novel Type I Interferon-Mediated Mechanism
Philberta Y. Leung, Susan L. Stevens, Amy E.B. Packard, Nikola S. Lessov, Tao Yang, Valerie K. Conrad, Noortje N.A.M. van den Dungen, Roger P. Simon and Mary P. Stenzel-Poore

Stroke. 2012;43:1383-1389, originally published April 23, 2012
https://doi.org/10.1161/STROKEAHA.111.641522

Download PDF
You have access
Toll-Like Receptor 7 Preconditioning Induces Robust Neuroprotection Against Stroke by a Novel Type I Interferon-Mediated Mechanism
Philberta Y. Leung, Susan L. Stevens, Amy E.B. Packard, Nikola S. Lessov, Tao Yang, Valerie K. Conrad, Noortje N.A.M. van den Dungen, Roger P. Simon, Mary P. Stenzel-Poore

Stroke May 2012, 43 (5) 1383-1389; DOI: https://doi.org/10.1161/STROKEAHA.111.641522

Figure 1.
Gardiquimod (GDQ) preconditioning reduces ischemic injury. A, C57Bl/6 mice were preconditio...
Show More

Gardiquimod (GDQ) preconditioning reduces ischemic injury. A, C57Bl/6 mice were preconditioned with escalating doses of GDQ (N=8; 10, 20, or 40 μg per mouse, subcutaneous) or saline (N=5) 72 hours before 60 minutes of middle cerebral artery occulusion (MCAO). Infarct size was determined 24 hours after MCAO. B, C57Bl/6 mice were pretreated with GDQ (N=8; 20 μg/mouse, subcutaneous) or saline (n=6) 72 hours before MCAO. Infarct size was determined 72 hours after MCAO. C, C57Bl/6 mice were pretreated with GDQ (N=5–6; 20 μg/mouse, subcutaneous) or saline (n=6) at various times before MCAO. Infarct size was determined 24 hours after MCAO. Two-way analysis of variance (ANOVA), Bonferroni post hoc, *P<0.01, **P<0.01, ***P<0.001 vs saline controls. D and E, C57BL/6 mice were treated with GDQ (N=5; 40 μg/mouse, subcutaneous) or saline (N=6) 72 hours before MCAO (60 minutes). Mice were then examined using the (D) neurological score (focal and general) and (E) corner test to determine neurological and sensorimotor deficits 24 hours after MCAO. Student t test: *P<0.01, **P<0.01, ***P<0.001 vs saline controls.
Show Less
You have access
Toll-Like Receptor 7 Preconditioning Induces Robust Neuroprotection Against Stroke by a Novel Type I Interferon-Mediated Mechanism
Philberta Y. Leung, Susan L. Stevens, Amy E.B. Packard, Nikola S. Lessov, Tao Yang, Valerie K. Conrad, Noortje N.A.M. van den Dungen, Roger P. Simon, Mary P. Stenzel-Poore

Stroke May 2012, 43 (5) 1383-1389; DOI: https://doi.org/10.1161/STROKEAHA.111.641522

Figure 2.
Gardiquimod (GDQ)-induced neuroprotection is mediated through Toll-like receptor (TLR) 7. TLR7^+/+...
Show More

Gardiquimod (GDQ)-induced neuroprotection is mediated through Toll-like receptor (TLR) 7. TLR7^+/+ (N=7) or TLR7^−/− (N=5–7) mice were preconditioned with GDQ (40 μg/mouse, subcutaneous) or saline 72 hours before middle cerebral artery occlusion (MCAO; 45 minutes). Infarct size was determined 24 hours after MCAO. Two-way analysis of variance (ANOVA), Bonferroni post hoc, **P<0.01 vs saline control for respective genotype.
Show Less