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Dietary phytol reduces clinical symptoms in experimental autoimmune encephalomyelitis (EAE) at least partially by modulating NOX2 expression

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Abstract

Multiple sclerosis (MS) is an inflammatory, demyelinating disease of the central nervous system. We investigated the effect of phytol in an animal model of MS, experimental autoimmune encephalomyelitis (EAE), as phytol, a plant-derived diterpene alcohol, exerts anti-inflammatory and redox-protective actions. We observed a significant amelioration of clinical symptoms in EAE C57BL/6N mice fed prophylactically with a phytol-enriched diet. Demyelination, DNA damage, and infiltration of immune cells, specifically TH1 cells, into the central nervous system were reduced in phytol-fed EAE mice. Furthermore, phytol reduced T-cell proliferation ex vivo. Phytanic acid — a metabolite of phytol — also reduced T-cell proliferation, specifically that of TH1 cells. Additionally, phytol-enriched diet increased the mRNA expression of nicotinamide adenine dinucleotide phosphate oxidase (NOX) 2 in white blood cells in the lymph nodes. Accordingly, phytol lost its anti-inflammatory effects in chimeric EAE C57BL/6N mice whose peripheral cells lack NOX2, indicating that phytol mediates its effects in peripheral cells via NOX2. Moreover, the effects of phytol on T-cell proliferation were also NOX2-dependent. In contrast, the T-cell subtype alterations and changes in proliferation induced by phytanic acid, the primary metabolite of phytol, were NOX2-independent. In conclusion, phytol supplementation of the diet leads to amelioration of EAE pathology in both a NOX2-dependent and a NOX2-independent manner via yet unknown mechanisms.

Key messages

  • Phytol diet ameliorates EAE pathology.

  • Phytol diet reduces demyelination, immune cell infiltration, and T-cell proliferation.

  • Phytol diet increases NOX2 mRNA expression in white blood cells in the lymph nodes.

  • Phytol mediates its effects in peripheral cells via NOX2.

  • Effects of phytol on T-cell proliferation were NOX2-dependent.

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Abbreviations

8-OHG:

8-Hydroxyguanosine

AF:

Alexa fluor

ANOVA:

Analysis of variance

APC:

Antigen-presenting cell

AUC:

Area under the curve

BMT:

Bone marrow transition

BrdU:

5-Bromo-2′-deoxyuridine

CFA:

Complete Freund’s adjuvant

CNS:

Central nervous system

CT:

Cycle threshold

DAPI:

4′,6-Diamidin-2-phenylindol

EAE:

Experimental autoimmune encephalomyelitis

FCS:

Fetal calf serum

FM:

Fluoromyelin

IFN:

Interferon

IL:

Interleukin

MACS:

Magnetic activated cell sorting

MOG:

Myelin oligodendrocyte protein

MRM:

Multiple reaction monitoring

MS:

Multiple sclerosis

NADPH:

Nicotinamide adenine dinucleotide phosphate

NOX2:

NADPH oxidase 2

PBS:

Phosphate-buffered saline

PCR:

Polymerase chain reaction

PH:

Phytol

PPIA:

Peptidylpropyl isomerase A

PTX:

Pertussis toxin

ROS:

Reactive oxygen species

RR-MS:

Relapse-remitting multiple sclerosis

SEM:

Standard error of the mean

SPF:

Specific pathogen-free

ST:

Standard

WBCs:

White blood cells

WT:

Wild type

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Acknowledgments

This work was supported by the Landesoffensive zur Entwicklung wissenschaftlich-ökonomischer Exzellenz (LOEWE) and the Zentrum: Translationale Medizin und Pharmakologie.

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Author notes

  1. Leonard Blum and Nadja Tafferner contributed equally to this work.

Authors and Affiliations

  1. pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe-University Hospital Frankfurt, Theodor-Stern-Kai 7, 60590, Frankfurt/Main, Germany

    Leonard Blum, Gerd Geisslinger & Susanne Schiffmann

  2. Branch for Translational Medicine and Pharmacology TMP, Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Theodor-Stern-Kai 7, 60596, Frankfurt am Main, Germany

    Nadja Tafferner, Ilknur Spring, Jennifer Kurz, Natasja deBruin, Michael J. Parnham & Susanne Schiffmann

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  1. Leonard Blum
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  2. Nadja Tafferner
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Correspondence to Susanne Schiffmann.

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Conflict of interest

MJP has been a consultant to Leo Pharma a/s and Xellia Pharmaceuticals. All the other authors declare that they have no conflict of interest.

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Blum, L., Tafferner, N., Spring, I. et al. Dietary phytol reduces clinical symptoms in experimental autoimmune encephalomyelitis (EAE) at least partially by modulating NOX2 expression. J Mol Med 96, 1131–1144 (2018). https://doi.org/10.1007/s00109-018-1689-7

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  • DOI: https://doi.org/10.1007/s00109-018-1689-7

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