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N-Acetylglucosaminyltransferase V exacerbates murine colitis with macrophage dysfunction and enhances colitic tumorigenesis

  • Original Article—Alimentary Tract
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An Editorial to this article was published on 24 December 2015

Abstract

Background

Oligosaccharide structures and their alterations have important roles in modulating intestinal inflammation. N-Acetylglucosaminyltransferase V (GnT-V) is involved in the biosynthesis of N-acetylglucosamine (GlcNAc) by β1,6-branching on N-glycans and is induced in various pathologic processes, such as inflammation and regeneration. GnT-V alters host immune responses by inhibiting the functions of CD4+ T cells and macrophages. The present study aimed to clarify the role of GnT-V in intestinal inflammation using GnT-V transgenic mice.

Methods

Colitis severity was compared between GnT-V transgenic mice and wild-type mice. β1,6-GlcNAc levels were investigated by phytohemagglutinin-L4 lectin blotting and flow cytometry. We investigated phagocytosis of macrophages by measuring the number of peritoneal-macrophage-ingested fluorescent latex beads by flow cytometry. Cytokine production in the culture supernatant of mononuclear cells from the spleen, mesenteric lymph nodes, and bone-marrow-derived macrophages was determined by enzyme-linked immunosorbent assay. Clodronate liposomes were intravenously injected to deplete macrophages in vivo. Chronic-colitis-associated tumorigenesis was assessed after 9 months of repeated administration of dextran sodium sulfate (DSS).

Results

DSS-induced colitis and colitis induced by trinitrobenzene sulfonic acid were markedly exacerbated in GnT-V transgenic mice compared with wild-type mice. Production of interleukin-10 and phagocytosis of macrophages were significantly impaired in GnT-V transgenic mice compared with wild-type mice. Clodronate liposome treatment to deplete macrophages blocked the exacerbation of DSS-induced colitis and impairment of interleukin-10 production in GnT-V transgenic mice. Chronic-colitis-associated tumorigenesis was significantly increased in GnT-V transgenic mice.

Conclusions

Overexpression of GnT-V exacerbated murine experimental colitis by inducing macrophage dysfunction, thereby enhancing colorectal tumorigenesis.

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Abbreviations

BMDM:

Bone-marrow–derived macrophage

CLP:

Cecal ligation and puncture

DSS:

Dextran sodium sulfate

EmGFP:

Emerald green fluorescent protein

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

GFP:

Green fluorescent protein

β1,6-GlcNAc:

β1,6-N-Acetylglucosamine

GnT-III:

N-Acetylglucosaminyltransferase III

GnT-V:

N-Acetylglucosaminyltransferase V

IBD:

Inflammatory bowel disease

IL:

Interleukin

LPS:

Lipopolysaccharides

PBS:

Phosphate-buffered saline

PHA-L4 :

Phytohemagglutinin-L4

TNBS:

2,4,6-Trinitrobenzenesulfonic acid

TLR4:

Toll-like receptor 4

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Acknowledgments

We thank M. Terao from the Department of Dermatology, Osaka University Graduate School of Medicine, for great support and mentorship in the performance of this research. This work was supported by a Grant-in-Aid for Scientific Research (A), No. 21249038, and a Grant-in-Aid for Young Scientists (B), No. 22790643, from the Japan Society for the Promotion of Science, a grant from the Global COE program of Osaka University funded by the Ministry of Education, Culture, Sports, Science, and Technology of Japan, and a Grant-in-Aid from the Smoking Research Foundation.

Conflict of interest

The authors declare that they have no conflict of interest.

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    Authors and Affiliations

    1. Department of Gastroenterology and Hepatology, Osaka University Graduate School of Medicine, 2-2 K1, Yamadaoka, Suita, Osaka, 565-0871, Japan

      Shinichiro Shinzaki, Hideki Iijima, Shoichiro Kawai, Eri Shiraishi, Satoshi Hiyama, Takahiro Inoue, Yoshito Hayashi, Yoshihiro Kamada, Masahiko Tsujii & Tetsuo Takehara

    2. Department of Molecular Biochemistry and Clinical Investigation, Osaka University Graduate School of Medicine, 1-7, Yamadaoka, Suita, Osaka, 565-0871, Japan

      Mayuko Ishii, Hironobu Fujii, Kana Wakamatsu, Shinji Takamatsu, Yoshihiro Kamada & Eiji Miyoshi

    3. Department of Pathology, Osaka University Graduate School of Medicine, 2-2, Yamadaoka, Suita, Osaka, 565-0871, Japan

      Eiichi Morii

    4. Research Center for Ultra-High Voltage Electron Microscopy, Osaka University, 7-1, Mihogaoka, Ibaraki, Osaka, 567-0047, Japan

      Ryusuke Kuwahara

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    Correspondence to Eiji Miyoshi.

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    S. Shinzaki and M. Ishii contributed equally to the manuscript.

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    Shinzaki, S., Ishii, M., Fujii, H. et al. N-Acetylglucosaminyltransferase V exacerbates murine colitis with macrophage dysfunction and enhances colitic tumorigenesis. J Gastroenterol 51, 357–369 (2016). https://doi.org/10.1007/s00535-015-1119-3

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    • DOI: https://doi.org/10.1007/s00535-015-1119-3

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