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Oral TNFα Modulation Alters Neutrophil Infiltration, Improves Cognition and Diminishes Tau and Amyloid Pathology in the 3xTgAD Mouse Model.

Gabbita SP, Johnson MF, Kobritz N, Eslami P, Poteshkina A, Varadarajan S, Turman J, Zemlan F, Harris-White ME - PLoS ONE (2015)

Bottom Line: IDT significantly reduced paired helical filament tau and fibrillar amyloid accumulation.Flow cytometry of brain cell populations revealed that IDT increased the infiltrating neutrophil population while reducing TNFα expression in this population.IDT is a safe and effective TNFα and innate immune system modulator.

View Article: PubMed Central - PubMed

Affiliation: P2D Bioscience, Inc., Cincinnati, Ohio, United States of America.

ABSTRACT
Cytokines such as TNFα can polarize microglia/macrophages into different neuroinflammatory types. Skewing of the phenotype towards a cytotoxic state is thought to impair phagocytosis and has been described in Alzheimer's Disease (AD). Neuroinflammation can be perpetuated by a cycle of increasing cytokine production and maintenance of a polarized activation state that contributes to AD progression. In this study, 3xTgAD mice, age 6 months, were treated orally with 3 doses of the TNFα modulating compound isoindolin-1,3 dithione (IDT) for 10 months. We demonstrate that IDT is a TNFα modulating compound both in vitro and in vivo. Following long-term IDT administration, mice were assessed for learning & memory and tissue and serum were collected for analysis. Results demonstrate that IDT is safe for long-term treatment and significantly improves learning and memory in the 3xTgAD mouse model. IDT significantly reduced paired helical filament tau and fibrillar amyloid accumulation. Flow cytometry of brain cell populations revealed that IDT increased the infiltrating neutrophil population while reducing TNFα expression in this population. IDT is a safe and effective TNFα and innate immune system modulator. Thus small molecule, orally bioavailable modulators are promising therapeutics for Alzheimer's disease.

No MeSH data available.


Related in: MedlinePlus

Flow cytometric analysis of leukocytes recovered from the brains of Con and IDT-treated 3xTgAD mice.(A) Representative flow cytometry profiles of the cell surface markers and gating strategy are shown for Con and Low IDT brains. (B) The percentage of leukocytes (CD45Hi+), microglia (CD45Dim+), granulocytes (CD45Hi+/CD11b+/GR1+) and neutrophils (CD45Hi+/CD11b+/GR1+/1A8+) among all cells analyzed across the four treatment groups. One-way ANOVA of the neutrophil populations revealed a significant IDT effect (F2, 10 = 6.468, P = 0.0023). Both Low/Med and High IDT treatment groups significantly increased neutrophil infiltration into the brain relative to Con 3xTgAD brains (*p < 0.05; **p < 0.01).
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pone.0137305.g004: Flow cytometric analysis of leukocytes recovered from the brains of Con and IDT-treated 3xTgAD mice.(A) Representative flow cytometry profiles of the cell surface markers and gating strategy are shown for Con and Low IDT brains. (B) The percentage of leukocytes (CD45Hi+), microglia (CD45Dim+), granulocytes (CD45Hi+/CD11b+/GR1+) and neutrophils (CD45Hi+/CD11b+/GR1+/1A8+) among all cells analyzed across the four treatment groups. One-way ANOVA of the neutrophil populations revealed a significant IDT effect (F2, 10 = 6.468, P = 0.0023). Both Low/Med and High IDT treatment groups significantly increased neutrophil infiltration into the brain relative to Con 3xTgAD brains (*p < 0.05; **p < 0.01).

Mentions: Flow cytometry was used to quantify and phenotype blood-derived leukocytes and CNS-resident microglia in 3xTgAD mice. As there was no statistical difference between Low diet and Med diet flow cytometry data, those two groups were combined for analysis. The gating strategy used to evaluate various cell populations is shown in Fig 4A. A representative Con diet and Low IDT mouse are shown. IDT treatment at any dose did not significantly reduce the % of CD45Hi+(infiltrating leukocytes) or CD45Hi+/Cd11b+/GR1+ (granulocytes) but did increase CD45Hi+/Cd11b+/GR1+/1A8+ (neutrophils) and reduce (~12%) the CD45Dim+ (microglia) in both the Low/Med and High diet doses (Fig 4B). Fig 5 shows the data for TNFα expression in the same populations. Fig 5 row A shows the mean fluorescence intensity (MFI) for TNFα. Only the neutrophil population showed a decrease in TNFα MFI with increasing IDT dose. Fig 5 row B shows the percentage of the isolated populations that were TNFα+. The total number of neutrophils that were TNFα+ decreased with increasing IDT dose. Fig 5, graph C shows TNFα protein levels in whole cortex homogenates where the Low dose IDT was most effective, perhaps reflecting a reduction in neuronal and extracellular space levels of TNFα also with Low and Med dose IDT.


Oral TNFα Modulation Alters Neutrophil Infiltration, Improves Cognition and Diminishes Tau and Amyloid Pathology in the 3xTgAD Mouse Model.

Gabbita SP, Johnson MF, Kobritz N, Eslami P, Poteshkina A, Varadarajan S, Turman J, Zemlan F, Harris-White ME - PLoS ONE (2015)

Flow cytometric analysis of leukocytes recovered from the brains of Con and IDT-treated 3xTgAD mice.(A) Representative flow cytometry profiles of the cell surface markers and gating strategy are shown for Con and Low IDT brains. (B) The percentage of leukocytes (CD45Hi+), microglia (CD45Dim+), granulocytes (CD45Hi+/CD11b+/GR1+) and neutrophils (CD45Hi+/CD11b+/GR1+/1A8+) among all cells analyzed across the four treatment groups. One-way ANOVA of the neutrophil populations revealed a significant IDT effect (F2, 10 = 6.468, P = 0.0023). Both Low/Med and High IDT treatment groups significantly increased neutrophil infiltration into the brain relative to Con 3xTgAD brains (*p < 0.05; **p < 0.01).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4593589&req=5

pone.0137305.g004: Flow cytometric analysis of leukocytes recovered from the brains of Con and IDT-treated 3xTgAD mice.(A) Representative flow cytometry profiles of the cell surface markers and gating strategy are shown for Con and Low IDT brains. (B) The percentage of leukocytes (CD45Hi+), microglia (CD45Dim+), granulocytes (CD45Hi+/CD11b+/GR1+) and neutrophils (CD45Hi+/CD11b+/GR1+/1A8+) among all cells analyzed across the four treatment groups. One-way ANOVA of the neutrophil populations revealed a significant IDT effect (F2, 10 = 6.468, P = 0.0023). Both Low/Med and High IDT treatment groups significantly increased neutrophil infiltration into the brain relative to Con 3xTgAD brains (*p < 0.05; **p < 0.01).
Mentions: Flow cytometry was used to quantify and phenotype blood-derived leukocytes and CNS-resident microglia in 3xTgAD mice. As there was no statistical difference between Low diet and Med diet flow cytometry data, those two groups were combined for analysis. The gating strategy used to evaluate various cell populations is shown in Fig 4A. A representative Con diet and Low IDT mouse are shown. IDT treatment at any dose did not significantly reduce the % of CD45Hi+(infiltrating leukocytes) or CD45Hi+/Cd11b+/GR1+ (granulocytes) but did increase CD45Hi+/Cd11b+/GR1+/1A8+ (neutrophils) and reduce (~12%) the CD45Dim+ (microglia) in both the Low/Med and High diet doses (Fig 4B). Fig 5 shows the data for TNFα expression in the same populations. Fig 5 row A shows the mean fluorescence intensity (MFI) for TNFα. Only the neutrophil population showed a decrease in TNFα MFI with increasing IDT dose. Fig 5 row B shows the percentage of the isolated populations that were TNFα+. The total number of neutrophils that were TNFα+ decreased with increasing IDT dose. Fig 5, graph C shows TNFα protein levels in whole cortex homogenates where the Low dose IDT was most effective, perhaps reflecting a reduction in neuronal and extracellular space levels of TNFα also with Low and Med dose IDT.

Bottom Line: IDT significantly reduced paired helical filament tau and fibrillar amyloid accumulation.Flow cytometry of brain cell populations revealed that IDT increased the infiltrating neutrophil population while reducing TNFα expression in this population.IDT is a safe and effective TNFα and innate immune system modulator.

View Article: PubMed Central - PubMed

Affiliation: P2D Bioscience, Inc., Cincinnati, Ohio, United States of America.

ABSTRACT
Cytokines such as TNFα can polarize microglia/macrophages into different neuroinflammatory types. Skewing of the phenotype towards a cytotoxic state is thought to impair phagocytosis and has been described in Alzheimer's Disease (AD). Neuroinflammation can be perpetuated by a cycle of increasing cytokine production and maintenance of a polarized activation state that contributes to AD progression. In this study, 3xTgAD mice, age 6 months, were treated orally with 3 doses of the TNFα modulating compound isoindolin-1,3 dithione (IDT) for 10 months. We demonstrate that IDT is a TNFα modulating compound both in vitro and in vivo. Following long-term IDT administration, mice were assessed for learning & memory and tissue and serum were collected for analysis. Results demonstrate that IDT is safe for long-term treatment and significantly improves learning and memory in the 3xTgAD mouse model. IDT significantly reduced paired helical filament tau and fibrillar amyloid accumulation. Flow cytometry of brain cell populations revealed that IDT increased the infiltrating neutrophil population while reducing TNFα expression in this population. IDT is a safe and effective TNFα and innate immune system modulator. Thus small molecule, orally bioavailable modulators are promising therapeutics for Alzheimer's disease.

No MeSH data available.


Related in: MedlinePlus