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Structure of a β-glucan from Grifola frondosa and its antitumor effect by activating Dectin-1/Syk/NF-κB signaling

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Abstract

A soluble homogeneous β-glucan, GFPBW1, with a molecular mass of 300 kDa was purified from the fraction of the fruit bodies of Grifola frondosa extracted with 5 % NaOH. Using various methods, such as infrared spectroscopy, NMR, methylation and monosaccharide composition analysis, its structure was determined to be a β-D-(1-3)-linked glucan backbone with a single β-D-(1-6)-linked glucopyranosyl residue branched at C-6 on every third residue. It induced TNF-α and IL-6 production and the activation of Syk and NF-κB signaling in resident peritoneal macrophages from ICR mice, which could be significantly inhibited by the blocking reagent laminarin. A competitive phagocytosis assay with FITC-zymosan indicated that GFPBW1 could bind to DC-associated C-type lectin 1 (Dectin-1). The TNF-α secretion and activation of Syk/NF-κB signaling triggered by GFPBW1 were enhanced in RAW264.7 cells overexpressing wild but not mutant (Δ38 and Y15S) Dectin-1. Furthermore, GFPBW1 potentiated the Concanavalin A-induced proliferative response of splenocytes and inhibited Sarcoma-180 growth allografted in ICR mice but not in immunodeficient BALB/c nu/nu mice. These results suggested that the antitumor activity of GFPBW1 was partially associated with the activation of macrophages via the Dectin-1/Syk/NF-κB signaling pathway. This molecule could be a promising biological response modifier with clear application for antitumor therapies.

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Abbreviations

BRMs:

Biological response modifiers

CARD9:

Caspase recruitment domain 9

ConA:

Concanavalin A

CR3:

Complement receptor 3

CTX:

Cyclophosphamide

DCs:

Dendritic cells

Dectin-1:

DC-associated C-type lectin 1

DMSO:

Dimethyl sulfoxide

FACS:

Fluorescence-activated cell sorting

GC:

Gas chromatography

G. frondosa :

Grifola frondosa (Fr.) S. F. Gray

GRN:

Grifolan

HPGPC:

High performance gel permeation chromatography

HRP:

Horseradish peroxidase

HSQC:

Heteronuclear single quantum coherence

IR:

Infrared

ITAM:

Immunoreceptor tyrosine-based activation motif

LPS:

Lipopolysaccharide

MS:

Mass spectrometry

NF-κB:

Nuclear factor κB

NMR:

Nuclear magnetic resonance

PCR:

Polymerase chain reaction

PMB:

Polymyxin B

POD:

Peroxidase

PRRs:

Pattern recognition receptors

Syk:

Spleen tyrosine kinase

TFA:

Trifluoroacetic acid

TLR2:

Toll like receptor 2

SDS:

Sodium dodecyl sulfate

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Acknowledgements

This work was supported by New Drug Creation and Manufacturing Program (2012ZX09301001-003), National Science Fund for Distinguished Young Scholars (81125025) and the funds for Industry-University-Research Institution Alliance in Guangdong Province, China (2010A090200041).

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Authors and Affiliations

  1. Glycochemistry & Glycobiology Lab, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Zhangjiang High-Tech Park, Shanghai, 201203, China

    Jianping Fang, Ying Wang, Xiaofen Lv, Xiaokun Shen, Xinyan Ni & Kan Ding

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  1. Jianping Fang
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  2. Ying Wang
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  3. Xiaofen Lv
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  4. Xiaokun Shen
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  6. Kan Ding
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Correspondence to Kan Ding.

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Jianping Fang and Ying Wang contributed equally to this work.

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Fang, J., Wang, Y., Lv, X. et al. Structure of a β-glucan from Grifola frondosa and its antitumor effect by activating Dectin-1/Syk/NF-κB signaling. Glycoconj J 29, 365–377 (2012). https://doi.org/10.1007/s10719-012-9416-z

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  • DOI: https://doi.org/10.1007/s10719-012-9416-z

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