2024/09/20 更新

写真a

マエダ メグミ
前田 恵
MAEDA Megumi
所属
環境生命自然科学学域 准教授
職名
准教授
外部リンク

学位

  • 農学 ( 岡山大学 )

研究キーワード

  • サイトカイン

  • アスベスト

  • Th2免疫応答

  • 抗腫瘍免疫

  • レクチン

  • N-グリカン

  • 植物抗原性糖鎖

  • 花粉アレルゲン

  • 食物アレルゲン

  • 糖鎖ポリマー

研究分野

  • ライフサイエンス / 応用生物化学

  • ライフサイエンス / 免疫学

学歴

  • 岡山大学    

    2003年4月 - 2006年3月

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    国名: 日本国

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  • 岡山大学    

    2001年4月 - 2003年3月

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    国名: 日本国

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  • 岡山大学   Faculty of Agriculture   Department of Agricultural Sciences

    1997年4月 - 2001年3月

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    国名: 日本国

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経歴

  • 岡山大学 学術研究院 環境生命自然科学学域 准教授

    2023年4月 - 現在

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  • 岡山大学 学術研究院 環境生命科学学域 准教授

    2021年4月 - 2023年3月

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  • 岡山大学大学院 環境生命科学研究科 准教授

    2017年3月 - 2021年3月

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  • ベルギー国ゲント大学 博士研究員

    2016年4月 - 2016年10月

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  • 岡山大学大学院 環境生命科学研究科 助教

    2014年4月 - 2017年2月

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  • ベルギー国ゲント大学 博士研究員

    2013年6月 - 2018年8月

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  • ベルギー国ゲント大学 博士研究員

    2012年8月 - 2013年1月

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  • 岡山大学大学院 環境生命科学研究科 助教(特任,WTT)

    2012年4月 - 2014年3月

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  • 岡山大学大学院 自然科学研究科 助教(特任,WTT)

    2011年4月 - 2012年3月

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  • 川崎医科大学医学部 応用医学 衛生学 助教

    2007年4月 - 2011年3月

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  • 川崎医科大学   School of Medicine

    2006年4月 - 2007年3月

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▼全件表示

所属学協会

  • 日本糖質学会

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  • 社団法人 日本生化学会

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  • 社団法人 日本農芸化学会

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  • 特定非営利活動法人 日本免疫学会

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委員歴

  • 第42回日本糖質学会 (鳥取)   Advisory Board  

    2023年2月 - 2023年9月   

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  • 第62回日本生化学会中国・四国支部例会 実行委員  

    2021年11月   

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論文

  • Construction of tomato plants with suppressed endo-β-N-acetylglucosaminidase activity using CRISPR-Cas9 mediated genome editing. 査読 国際誌

    Naoko Okamoto, Megumi Maeda, Chiharu Yamamoto, Reo Kodama, Koichi Sugimoto, Yoshihito Shinozaki, Hiroshi Ezura, Yoshinobu Kimura

    Plant physiology and biochemistry : PPB   190   203 - 211   2022年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    High mannose-type free N-glycans with a single N-acetyl-D-glucosamine (GlcNAc) residue at the reducing end (GN1-HMT-FNGs) are produced by cytosolic endo-β-N-acetylglucosaminidase (EC:3.2.1.96) (ENGase) and are ubiquitous in differentiating and growing plant cells. To elucidate the physiological functions of HMT-FNGs in plants, we identified the ENGase gene in tomato (Solyc06g050930) and detected ENGase activity and increased production of GN1-HMT-FNGs during tomato fruit maturation. However, the precise role of GN1-HMT-FNGs in fruit maturation remains unclear. In this study, we established tomato ENGase mutants with suppressed ENGase activity via CRISPR/Cas9 genome editing technology. DNA sequencing of the Δeng mutants (T0 and T1 generations) revealed that they had the same mutations in the genomic DNA around the target sequences. Three null CRISPR/Cas9 segregant plants of the T1 generation (Δeng1-2, -22, and -26) were used to measure ENGase activity and analyze the structural features of HMT-FNGs in the leaves. The Δeng mutants did not exhibit ENGase activity and produced GN2-HMT-FNGs bearing tow GlcNAc residues at the reducing end side instead of GN1-HMT-FNGs. The Δeng mutants lack the N-terminal region of ENGase, indicating that the N-terminal region is important for full ENGase activity. The fruits of Δeng mutants (T2 generation) also showed loss of ENGase activity and similar structural features of HMT-FNGs of the T1 generation. However, there was no significant difference in fruit maturation between the T2 generation of the Δeng mutants and the wild type. The Δeng mutants rich in GN2-HMT-FNGs could be offered as a new tomato that is different from wild type containing GN1-HMT-FNGs.

    DOI: 10.1016/j.plaphy.2022.08.009

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  • Structural features of free N-glycans in α1,3/4-fucosidase-deficient Arabidopsis thaliana: deletion of α1,3/4-fucosidase activity induced accumulation of plant complex type GN1 free N-glycans. 査読 国際誌

    Shun Takata, Megumi Hayashi, Megumi Maeda, Takeshi Ishimizu, Yoshinobu Kimura

    Bioscience, biotechnology, and biochemistry   86 ( 10 )   1413 - 1416   2022年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Deletion of α-1,3/4-fucosidase activity in Arabidopsis thaliana resulted in the accumulation of GN1-type free N-glycans with the Lewis a epitope (GN1-FNG). This suggests that the release of α-fucose residue(s) may trigger rapid degradation of the plant complex-type (PCT) GN1-FNG. The fact that PCT-GN1-FNG has rarely been detected to date is probably due to its easier degradation compared with PCT-GN2-FNG.

    DOI: 10.1093/bbb/zbac120

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  • Improved method for preparation and purification of recombinant α-synuclein: high-mannose-type free N-glycan prepared from an edible bean (Vigna angulari, Azuki bean) inhibits α-synuclein aggregation. 査読 国際誌

    Shota Kosaka, Makoto Katsube, Megumi Maeda, Yoshinobu Kimura

    Bioscience, biotechnology, and biochemistry   86 ( 6 )   770 - 774   2022年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Parkinson's disease is characterized by the accumulation of amyloid, which consists of α-synuclein (α-Syn). To screen compounds with amyloid aggregation inhibitory activity, an effective method for the preparation of α-Syn is a prerequisite. We established a simpler method for α-Syn preparation using freeze-thaw treatment of transformed Escherichia coli. Furthermore, we found that the high-mannose type free N-glycans could prevent α-Syn aggregation.

    DOI: 10.1093/bbb/zbac040

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  • Improved assay system for acidic peptide: N-glycanase (aPNGase) activity in plant extracts. 査読 国際誌

    Chiharu Yamamoto, Mikako Ogura, Ryota Uemura, Maeda Megumi, Hiroyuki Kajiura, Ryo Misaki, Kazuhito Fujiyama, Yoshinobu Kimura

    Analytical biochemistry   634   114367 - 114367   2021年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Plant acidic peptide: N-glycanase (aPNGase) release N-glycans from glycopeptides during the degradation process of glycoproteins in developing or growing plants. We have previously developed a new method to detect the aPNGase activity in crude extracts, which is prerequisite for the construction of aPNGase knockout or overexpression lines. However, this method has the disadvantage of requiring de-sialylation treatment and a lectin chromatography. In this study, therefore, we improved the simple and accurate method for detecting aPNGase activity using anion-exchange HPLC requiring neither the desialylation treatment nor the lectin affinity chromatography.

    DOI: 10.1016/j.ab.2021.114367

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  • Large-scale preparation of sialyl-Tn antigen-containing peptides from mucin-like glycoproteins in boar seminal gel. 査読 国際誌

    Ryota Takeuchi, Megumi Maeda, Miran Nakano, Hiroaki Funahashi, Yoshinobu Kimura

    Bioscience, biotechnology, and biochemistry   85 ( 9 )   2022 - 2025   2021年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Sialyl-Tn antigen, a tumor antigen, is a valuable ligand for the purification of proteins that specifically bind to it. Here, we developed a new method for the preparation of large amounts of sialyl-Tn antigen-containing peptides from an unused resource, boar seminal gel. The glycopeptides were prepared from the actinase E digests by a combination of gel filtration and hydrophilic partitioning.

    DOI: 10.1093/bbb/zbab114

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  • Purification, Characterization, and Gene Expression of Rice Endo-β-N-Acetylglucosaminidase, Endo-Os 査読 国際誌

    Megumi Maeda, Naoko Okamoto, Norie Araki, Yoshinobu Kimura

    Frontiers in Plant Science   12   647684 - 647684   2021年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Frontiers Media SA  

    In the endoplasmic reticulum-associated degradation system of plant and animal cells, high-mannose type free <italic>N</italic>-glycans (HMT-FNGs) are produced from misfolded glycoproteins prior to proteasomal degradation, and two enzymes, cytosolic peptide:<italic>N</italic>-glycanase (cPNGase) and endo-β-<italic>N-</italic>acetylglucosaminidase (endo-β-GlcNAc-ase), are involved in the deglycosylation. Although the physiological functions of these FNGs in plant growth and development remain to be elucidated, detailed characterization of cPNGase and endo-β-GlcNAc-ase is required. In our previous work, we described the purification, characterization, and subcellular distribution of some plant endo-β-GlcNAc-ases and preliminarily reported the gene information of rice endo-β-GlcNAc-ase (Endo-Os). Furthermore, we analyzed the changes in gene expression of endo-β-GlcNAc-ase during tomato fruit maturation and constructed a mutant line of <italic>Arabidopsis thaliana</italic>, in which the two endo-β-GlcNAc-ase genes were knocked-out based on the Endo-Os gene. In this report, we describe the purification, characterization, amino acid sequence, and gene cloning of Endo-Os in detail. Purified Endo-Os, with an optimal pH of 6.5, showed high activity for high-mannose type <italic>N</italic>-glycans bearing the Manα1-2Manα1-3Manβ1 unit; this substrate specificity was almost the same as that of other plant endo-β-GlcNAc-ases, suggesting that Endo-Os plays a critical role in the production of HTM-FNGs in the cytosol. Electrospray ionization-mass spectrometry analysis of the tryptic peptides revealed 17 internal amino acid sequences, including the C terminus; the N-terminal sequence could not be identified due to chemical modification. These internal amino acid sequences were consistent with the amino acid sequence (UniProt ID: Q5W6R1) deduced from the <italic>Oryza sativa</italic> cDNA clone AK112067 (gene ID: <italic>Os05g0346500</italic>). Recombinant Endo-Os expressed in <italic>Escherichia coli</italic> using cDNA showed the same enzymatic properties as those of native Endo-Os.

    DOI: 10.3389/fpls.2021.647684

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  • Direct evidence of cytosolic PNGase activity in Arabidopsis thaliana: in vitro assay system for plant cPNGase activity 査読 国際誌

    Sahoko Shirai, Ryota Uemura, Megumi Maeda, Hiroyuki Kajiura, Ryo Misaki, Kazuhito Fujiyama, Yoshinobu Kimura

    Bioscience, Biotechnology, and Biochemistry   85 ( 6 )   1460 - 1463   2021年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Oxford University Press (OUP)  

    <title>ABSTRACT</title>
    Cytosolic peptide:N-glycanase (cPNGase), which occurs ubiquitously in eukaryotic cells, is involved in the de-N-glycosylation of misfolded glycoproteins in the protein quality control system. In this study, we aimed to provide direct evidence of plant cPNGase activity against a denatured glycoprotein using a crude extract prepared from a mutant line of Arabidopsis thaliana lacking 2 acidic PNGase genes.

    DOI: 10.1093/bbb/zbab047

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    その他リンク: http://academic.oup.com/bbb/article-pdf/85/6/1460/38192129/zbab047.pdf

  • Purification and molecular characterization of a truncated-type Ara h 1, a major peanut allergen: oligomer structure, antigenicity, and glycoform. 査読 国際誌

    Asaduzzaman Md, Megumi Maeda, Teruaki Matsui, Yoshihiro Takasato, Komei Ito, Yoshinobu Kimura

    Glycoconjugate journal   38 ( 1 )   67 - 76   2021年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Peanut allergies are among the most severe food allergies, and several allergenic proteins referred to as Ara h 1-Ara h 17 have been identified from peanut seeds. The molecular characterization of Ara h 1 (63 kDa), a glycosylated allergen, has almost been completed, and the occurrence of two homologous genes (clone 41B and clone P17) has been identified. In this study, we found a new variant of Ara h 1 i.e. 54 kDa, in which the N-terminal amino acid sequence was EGREGEQ-, indicating that the N-terminal domain of 63 kDa Ara h 1 had been removed. This new isoform was obtained from the run-through fraction of hydrophobic interaction chromatography while 63 kDa Ara h 1 was tightly bound to the hydrophobic resins, suggesting that the removal of the N-terminal domain resulted in extreme hydrophilic properties. We found that 63 kDa Ara h 1 occurs as higher order homo-oligomeric conformations such as decamer or nonamer, while 54 kDa Ara h 1 occurs exclusively as a homotrimer, indicating that the N-terminal domain of the 63 kDa molecule may be involved in higher order oligomerization. When antisera from peanut-allergic patients were treated with both the Ara h 1 molecules, the immunoglobulin E (IgE) antibodies in these sera reacted with each Ara h 1 molecule, suggesting that the C-terminal as well as the N-terminal domains of Ara h 1 contribute significantly to the epitope formations of this peanut glycoallergen. Furthermore, the glycoform analyses of N-glycans linked to 63 kDa and 54 kDa Ara h 1 subunits revealed that both typical high-mannose type and β-xylosylated type N-glycans are linked to the molecules. The cross-reactivity of IgE against Ara h 1 in the serum of one peanut allergy patient was completely lost by de-N-glycosylation, indicating the N-glycan of Ara h 1 was the sole epitope for the Ara h 1- specific IgE in the patient.

    DOI: 10.1007/s10719-020-09969-1

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  • Effect of asbestos exposure on differentiation and function of cytotoxic T lymphocytes. 査読 国際誌

    Naoko Kumagai-Takei, Yasumitsu Nishimura, Megumi Maeda, Hiroaki Hayashi, Hidenori Matsuzaki, Suni Lee, Kei Yoshitome, Tatsuo Ito, Takemi Otsuki

    Environmental health and preventive medicine   25 ( 1 )   59 - 59   2020年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Asbestos exposure is known to cause malignant mesothelioma, which is associated with poor prognosis. We focused on and examined the effect of asbestos exposure on the differentiation and function of cytotoxic T lymphocytes (CTLs). CTLs have the ability to specifically attack tumor cells after being differentiated from naïve CD8+ T cells following antigen stimulation. Exposure to chrysotile B asbestos suppressed the differentiation of CTLs during the mixed lymphocyte reaction (MLR) and was associated with a decrease in proliferation of CD8+ T cells. Additionally, in an effort to investigate the mechanism associated with suppressed CTL differentiation upon exposure to asbestos, we focused on IL-2, a cytokine involved in T cell proliferation. Our findings indicated that insufficient levels of IL-2 are not the main cause for the suppressed induction of CTLs by asbestos exposure, although they suggest potential improvement in the suppressed CTL function. Furthermore, the functional properties of peripheral blood CD8+ lymphocytes from asbestos-exposed individuals with pleural plaque (PP) and patients with malignant mesothelioma (MM) were examined. MM patients showed lower perforin levels in CD8+ lymphocytes following stimulation compared with PP-positive individuals. The production capacity of IFN-γ in the MM group tended to be lower compared with healthy volunteers or PP-positive individuals. In an effort to determine whether chronic and direct asbestos exposure affected the function of CD8+ T cells, cultured human CD8+ T cells were employed as an in vitro model and subjected to long-term exposure to chrysotile (CH) asbestos. This resulted in decreased levels of intracellular perforin and secreted IFN-γ. Those findings underlie the possibility that impaired CD8+ lymphocyte function is caused by asbestos exposure, which fail to suppress the development of MM. Our studies therefore reveal novel effects of asbestos exposure on CTLs, which might contribute towards the development and implementation of an effective strategy for the prevention and cure of malignant mesothelioma.

    DOI: 10.1186/s12199-020-00900-6

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  • Convenient preparation of an antigenic oligosaccharide from white kidney bean powder: A useful plant oligosaccharide for synthesis of immunoactive glycopolymer. 査読 国際誌

    Mariko Kimura, Mikako Ogura, Miyuki Akamatsu, Kaede Sugimoto, Megumi Maeda, Teruhiko Nitoda, Haruko Nagasawa-Fujimori, Hirotaka Yamashita, Yoshinobu Kimura

    International journal of biological macromolecules   153   1016 - 1023   2020年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Plant glycoproteins, especially allergenic glycoproteins such as pollen allergens, often carry antigenic N-glycans with α1-3 fucose and/or β1-2 xylose residue(s) on the trimannosyl core structure. We previously reported that one of such antigenic free-form N-glycans, Man3Xyl1Fuc1GlcNAc2 (M3FX) suppressed IL-4 production from Th2 cells of pollinosis patients. For the molecular-level analysis of this immunoactivity, an effective and convenient procedure for large scale preparation of the immunoactive free-form N-glycan and a synthesis of glycopolymers bearing multivalent M3FX has been required. During the preparation of prebiotic oligosaccharides from several edible beans, we found that the free-form M3FX accumulates in relatively large amounts in white kidney beans. In this report, we describe a new procedure for preparation of M3FX from white kidney bean powders by a combination of ion-exchange method, gel-filtration, and hydrophilic partitioning. The high-purity of M3FX prepared by this procedure was confirmed by MS-analysis and 1H-NMR, suggesting that the free-form M3FX can be used for the synthesis of neoglycopolymer. Using this new procedure, the immunoactive oligosaccharide can be prepared without the chemical method such as hydrazinolysis and other purification steps required to exclude other type of N-glycans.

    DOI: 10.1016/j.ijbiomac.2019.10.231

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  • Synthesis and preliminary evaluation of neoglycopolymers carrying multivalent N-glycopeptide units. 査読 国際誌

    Naoto Takeda, Megumi Maeda, Satsuki Itano, Miho Takase, Mariko Kimura, Yoshinobu Kimura

    International journal of biological macromolecules   147   1294 - 1300   2020年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In the present study, for the discovery of uncharacterized glycan-binding receptors or lectin-like receptors in plants, we developed neoglycopolymers to which three types of N-glycopeptides are conjugated; the first with plant complex type N-glycan (M3FX), the second with high-mannose type N-glycan (M8), and the third with animal complex type N-glycan (NeuAc2Gal2GN2M3). Three types of Asn-oligosaccharide (Asn-M3FX, Asn-M8, or Asn-NeuAc2Gal2GN2M3) were prepared from storage glycoproteins of Ginkgo biloba seeds, Vigna angularis seeds, and egg yolk glycopeptides from actinase digests of each glycoproteins or glycopeptide. Neoglycopolymers were synthesized such that the α-amino groups of Asn-oligosaccharide were coupled to the carboxyl groups of poly-γ-L-glutamic acid (γ-L-PGA) with 4-(4,6-Dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride n-hydrate (DMT-MM). The resulting neoglycopolymers were purified through a combination of gel-filtration and reverse-phase HPLC. The incorporation of N-glycans into γ-L-PGA (mol%) was estimated through amino acid composition analysis after acid hydrolysis. The incorporation rates of Asn-M3FX, Asn-M8, and Asn-NeuAc2Gal2GN2M3 into γ-L-PGA were 15.4%, 8.6%, and 11.1%, indicating that nearly 890, 500, and 640 molecules of N-glycans were conjugated with γ-L-PGA, respectively. Furthermore, we confirmed that the neoglycopolymer carrying the multivalent high-mannose type N-glycans is a useful tool for rapid purification of mannose-binding protein, Concanavalin A, from jack bean extract.

    DOI: 10.1016/j.ijbiomac.2019.09.255

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  • Cytosolic Free N-Glycans Are Retro-Transported Into the Endoplasmic Reticulum in Plant Cells. 査読 国際誌

    Makoto Katsube, Natsuki Ebara, Megumi Maeda, Yoshinobu Kimura

    Frontiers in plant science   11   610124 - 610124   2020年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    During endoplasmic reticulum (ER)-associated degradation, free N-glycans (FNGs) are produced from misfolded nascent glycoproteins via the combination of the cytosolic peptide N-glycanase (cPNGase) and endo-β-N-acetylglucosaminidase (ENGase) in the plant cytosol. The resulting high-mannose type (HMT)-FNGs, which carry one GlcNAc residue at the reducing end (GN1-FNGs), are ubiquitously found in developing plant cells. In a previous study, we found that HMT-FNGs assisted in protein folding and inhibited β-amyloid fibril formation, suggesting a possible biofunction of FNGs involved in the protein folding system. However, whether these HMT-FNGs occur in the ER, an organelle involved in protein folding, remained unclear. On the contrary, we also reported the presence of plant complex type (PCT)-GN1-FNGs, which carry the Lewisa epitope at the non-reducing end, indicating that these FNGs had been fully processed in the Golgi apparatus. Since plant ENGase was active toward HMT-N-glycans but not PCT-N-glycans that carry β1-2xylosyl and/or α1-3 fucosyl residue(s), these PCT-GN1-FNGs did not appear to be produced from fully processed glycoproteins that harbored PCT-N-glycans via ENGase activity. Interestingly, PCT-GN1-FNGs were found in the extracellular space, suggesting that HMT-GN1-FNGs formed in the cytosol might be transported back to the ER and processed in the Golgi apparatus through the protein secretion pathway. As the first step in elucidating the production mechanism of PCT-GN1-FNGs, we analyzed the structures of free oligosaccharides in plant microsomes and proved that HMT-FNGs (Man9-7GlcNAc1 and Man9-8GlcNAc2) could be found in microsomes, which almost consist of the ER compartments.

    DOI: 10.3389/fpls.2020.610124

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  • Plant complex type free N-glycans occur in tomato xylem sap. 査読 国際誌

    Yuta Tsujimori, Mikako Ogura, Md Ziaur Rahman, Megumi Maeda, Yoshinobu Kimura

    Bioscience, biotechnology, and biochemistry   83 ( 7 )   1310 - 1314   2019年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Free N-glycans (FNGs) are ubiquitous in growing plants. Further, acidic peptide:N-glycanase is believed to be involved in the production of plant complex-type FNGs (PCT-FNGs) during the degradation of dysfunctional glycoproteins. However, the distribution of PCT-FNGs in growing plants has not been analyzed. Here, we report the occurrence of PCT-FNGs in the xylem sap of the stem of the tomato plant. Abbreviations: RP-HPLC: reversed-phase HPLC; SF-HPLC: size-fractionation HPLC; PA-: pyridylamino; PCT: plant complex type; Hex: hexose; HexNAc: N-acetylhexosamine; Pen: pentose; Deoxyhex: deoxyhexose; Man: D-mannose; GlcNAc: N-acetyl-D-glucosamine; Xyl: D-xylose; Fuc: L-fucose; Lea: Lewis a (Galβ1-3(Fucα1-4)GlcNAc); PCT: plant complex type; M3FX: Manα1-6(Manα1-3)(Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc-PA; GN2M3FX: GlcNAcβ1-2Manα1-6(GlcNAcβ1-2Manα1-3)(Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc-PA; (Lea)1GN1M3FX: Galβ1-3(Fucα1-4)GlcNAc1-2 Manα1-6(GlcNAcβ1-2Manα1-3)(Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc-PA or GlcNAc1-2Manα1-6(Galβ1-3(Fucα1-4)GlcNAc1-2Manα1-3)(Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc-PA.

    DOI: 10.1080/09168451.2019.1608803

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  • アスベスト曝露によるヒト末梢血CD4+細胞からのIL-17産生誘導

    大槻 剛巳, 前田 恵, 李 順姫, 吉留 敬, 武井 直子, 西村 泰光

    産業衛生学雑誌   61 ( 臨増 )   444 - 444   2019年5月

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    記述言語:日本語   出版者・発行元:(公社)日本産業衛生学会  

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  • Aberrant expression of FoxP3 in a human T cell line possessing regulatory T cell‑like function and exposed continuously to asbestos fibers. 査読 国際誌

    Maeda M, Matsuzaki H, Yamamoto S, Lee S, Kumagai-Takei N, Yoshitome K, Min Y, Sada N, Nishimura Y, Otsuki T

    Oncology reports   40 ( 2 )   748 - 758   2018年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.3892/or.2018.6481

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  • Molecular characterization of second tomato α1,3/4-fucosidase (α-Fuc'ase Sl-2), a member of glycosyl hydrolase family 29 active toward the core α1,3-fucosyl residue in plant N-glycans. 査読 国際誌

    Md Ziaur Rahman, Yuta Tsujimori, Megumi Maeda, Md Anowar Hossain, Takeshi Ishimizu, Yoshinobu Kimura

    Journal of biochemistry   164 ( 1 )   53 - 63   2018年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In a previous study, we molecular-characterized a tomato (Solanum lycopersicum) α1, 3/4-fucosidase (α-Fuc'ase Sl-1) encoded in a tomato gene (Solyc03g006980), indicating that α-Fuc'ase Sl-1 is involved in the turnover of Lea epitope-containing N-glycans. In this study, we have characterized another tomato gene (Solyc11g069010) encoding α1, 3/4-fucosidase (α-Fuc'ase Sl-2), which is also active toward the complex type N-glycans containing Lea epitope(s). The baculovirus-insect cell expression system was used to express that α-Fuc'ase Sl-2 with anti-FLAG tag, and the expression product (rFuc'ase Sl-2), was found as a 65 kDa protein using SDS-PAGE and has an optimum pH of around 5.0. Similarly to rFuc'ase Sl-1, rFuc'ase Sl-2 hydrolyzed the non-reducing terminal α1, 3-fucose residue on LNFP III and α1, 4-fucose residues of Lea epitopes on plant complex type N-glycans, but not the core α1, 3-fucose residue on Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc or Fucα1-3GlcNAc. However, we found that both α-Fuc'ases Sl-1 and Sl-2 were specifically active toward α1, 3-fucose residue on GlcNAcβ1-4(Fucα1-3)GlcNAc, indicating that the non-substituted β-GlcNAc linked to the proximal GlcNAc residue of the core tri-saccharide moiety of plant specific N-glycans must be a pre-requisite for α-Fuc'ase activity. A 3 D modelled structure of the catalytic sites of α-Fuc'ase Sl-2 suggested that Asp192 and Glu236 may be important for binding to the α1, 3/4 fucose residue.

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  • Novel assay system for acidic Peptide:N-glycanase (aPNGase) activity in crude plant extract. 査読 国際誌

    Ryota Uemura, Mikako Ogura, Chihiro Matsumaru, Tsuyoshi Akiyama, Megumi Maeda, Yoshinobu Kimura

    Bioscience, biotechnology, and biochemistry   82 ( 7 )   1172 - 1175   2018年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Acidic peptide:N-glycanase (aPNGase) plays a pivotal role in plant glycoprotein turnover. For the construction of aPNGase-knockout or -overexpressing plants, a new method to detect the activity in crude plant extracts is required because endogenous peptidases present in the extract hamper enzyme assays using fluorescence-labeled N-glycopeptides as a substrate. In this study, we developed a new method for measuring aPNGase activity in crude extracts from plant materials.

    DOI: 10.1080/09168451.2018.1459464

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  • Inflammatory Alteration of Human T Cells Exposed Continuously to Asbestos. 査読 国際誌

    Kumagai-Takei N, Yamamoto S, Lee S, Maeda M, Masuzzaki H, Sada N, Yu M, Yoshitome K, Nishimura Y, Otsuki T

    International journal of molecular sciences   19 ( 2 )   2018年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.3390/ijms19020504

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  • Degradation pathway of plant complex-type n-glycans: Identification and characterization of a key α1,3-fucosidase from glycoside hydrolase family 29 査読 国際誌

    Shun Kato, Megumi Hayashi, Mai Kitagawa, Hiroyuki Kajiura, Megumi Maeda, Yoshinobu Kimura, Kiyohiko Igarashi, Masahiro Kasahara, Takeshi Ishimizu

    Biochemical Journal   475 ( 1 )   305 - 317   2018年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Portland Press Ltd  

    Plant complex-type N-glycans are characterized by the presence of α1,3-linked fucose towards the proximal N-acetylglucosamine residue and β1,2-linked xylose towards the β-mannose residue. These glycans are ultimately degraded by the activity of several glycoside hydrolases. However, the degradation pathway of plant complex-type N-glycans has not been entirely elucidated because the gene encoding α1,3-fucosidase, a glycoside hydrolase acting on plant complex-type N-glycans, has not yet been identified, and its substrate specificity remains to be determined. In the present study, we found that AtFUC1 (an Arabidopsis GH29 α-fucosidase) is an α1,3-fucosidase acting on plant complex-type N-glycans. This fucosidase has been known to act on α1,4-fucoside linkage in the Lewis A epitope of plant complex-type N-glycans. We found that this glycoside hydrolase specifically acted on GlcNAcβ1–4(Fucα1–3)GlcNAc, a degradation product of plant complex-type N-glycans, by sequential actions of vacuolar α-mannosi-dase, β1,2-xylosidase, and endo-β-mannosidase. The AtFUC1-deficient mutant showed no distinct phenotypic plant growth features
    however, it accumulated GlcNAcβ1–4 (Fucα1–3)GlcNAc, a substrate of AtFUC1. These results showed that AtFUC1 is an α1,3-fucosidase acting on plant complex-type N-glycans and elucidated the degradation pathway of plant complex-type N-glycans.

    DOI: 10.1042/BCJ20170106

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  • Skewing T helper cells exposed to asbestos fibers toward reduction of tumor immunity or activation of autoimmunity 査読

    Kumagai-Takei N, Lee S, Matsuzaki H, Maeda M, Yu M, Sada N, Yoshitome K, Nishimura Y, Otsuki T

    Kawasaki Med J   44 ( 4 )   33 - 40   2018年

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  • Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki) 査読 国際誌

    Toshihiro Osada, Megumi Maeda, Chinatsu Tanabe, Kaori Furuta, Christopher J. Vavricka, Eiji Sasaki, Mitsuhiro Okano, Yoshinobu Kimura

    CARBOHYDRATE RESEARCH   448   18 - 23   2017年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI LTD  

    Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. However, the structures of N-glycans linked to Cha o 3 remain to be determined. In this study, therefore, we analyzed the glycoform of Cha o 3 and found that this glycoallergen carries exclusively plant complex-type N-glycans; major structures were G1cNAc2_ Man(3)Xyl(1)Fuc(1)GlcNAc(2) (39%), GaliFuciGlcNAc(2)Man(3)Xyl(1)FuciGlcNAc(2) (14%), and Gal(2)Fuc(2)G(1)cNAc2Man3. Xyl(1)Fuc(1)GIcNAc(2) (25%). The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GIcNAc(2)Man3Xyl(1)Fuc(1)GIcNAc(2) is a common structural feature of glycoallergens from Cupressaceae pollens. (C) 2017 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.carres.2017.05.005

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  • Search for biomarkers of asbestos exposure and asbestos-induced cancers in investigations of the immunological effects of asbestos. 査読 国際誌

    Matsuzaki H, Kumagai-Takei N, Lee S, Maeda M, Sada N, Hatayama T, Yamamoto S, Ikeda M, Yoshitome K, Min Y, Nishimura Y, Otsuki T

    Environmental health and preventive medicine   22 ( 1 )   53 - 53   2017年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1186/s12199-017-0661-4

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  • Induction of IL-17 production from human peripheral blood CD4+ cells by asbestos exposure. 査読 国際誌

    Maeda M, Chen Y, Lee S, Kumagai-Takei N, Yoshitome K, Matsuzaki H, Yamamoto S, Hatayama T, Ikeda M, Nishimura Y, Otsuki T

    International journal of oncology   50 ( 6 )   2024 - 2032   2017年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.3892/ijo.2017.3991

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  • Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 involved in the degradation of plant complex type N-glycans 査読 国際誌

    Md. Ziaur Rahman, Megumi Maeda, Satsuki Itano, Md. Anowar Hossain, Takeshi Ishimizu, Yoshinobu Kimura

    Journal of Biochemistry   161 ( 5 )   421 - 432   2017年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Oxford University Press  

    In this study, we identified a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans. Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) was expressed using a baculovirus-insect cell expression system. rFuc'ase Sl-1 is 55 kDa in size and has an optimum pH around 4.5. It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. Molecular 3D modelling of α-Fuc'ase Sl-1 and structure/sequence interpretation based on comparison with a homologous a-fucosidase from Bifidobacterium longum subsp. infantis (Blon-2336) indicated that residues Asp193 and Glu237 might be important for substrate binding.

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  • アスベスト長期曝露ヒト制御性T細胞株モデルにおける細胞周期の解析

    大槻 剛巳, 李 順姫, 松崎 秀紀, 武井 直子, 吉留 敬, 西村 泰光, 前田 恵

    産業衛生学雑誌   59 ( 臨増 )   505 - 505   2017年5月

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    記述言語:日本語   出版者・発行元:(公社)日本産業衛生学会  

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  • Occurrence of complex type free N-glycans with a single GlcNAc residue at the reducing termini in the fresh-water plant, Egeria densa 査読 国際誌

    Megumi Maeda, Natsuki Ebara, Misato Tani, Christopher J. Vavricka, Yoshinobu Kimura

    Glycoconjugate Journal   34 ( 2 )   229 - 240   2017年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Springer New York LLC  

    In our previous study, we found unique free N-glycans (FNGs), which carry a single GlcNAc residue (GN1) at the reducing-end side and the Lewis-a epitope at the non-reducing-end side, in the culture broth of rice cells. Based on the FNG structural features and the substrate specificity of plant ENGase, we hypothesized that there might be a novel biosynthetic mechanism responsible for the production of these unique GN1-FNGs, in which high-mannose type (HMT)-GN1-FNGs produced in the cytosol from misfolded glycoproteins by ENGase are transported back into the endoplasmic reticulum and processed to plant complex type (PCT)-GN1-FNGs in the Golgi apparatus. Until now, however, PCT-GN1-FNGs had only been found in the culture broth of rice cultured cells and never in plants, suggesting that the formation of PCT-GN1-FNGs might be generated under special or artificial conditions. In this study, we confirm the presence of PCT-GN1-FNGs, HMT-GN1-FNGs and PCT-GN2-FNGs in the fresh-water plant Egeria densa. These results suggest that a mechanism responsible for the production of PCT-GN1-FNG is present in native plant tissues.

    DOI: 10.1007/s10719-016-9758-z

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  • Accelerated cell cycle progression of human regulatory T cell-like cell line caused by continuous exposure to asbestos fibers. 査読 国際誌

    Lee S, Matsuzaki H, Maeda M, Yamamoto S, Kumagai-Takei N, Hatayama T, Ikeda M, Yoshitome K, Nishimura Y, Otsuki T

    International journal of oncology   50 ( 1 )   66 - 74   2017年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.3892/ijo.2016.3776

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  • Structural feature of N-glycans of bamboo shoot glycoproteins: useful source of plant antigenic N-glycans 査読 国際誌

    Chinatsu Tanabe, Kaori Furuta, Megumi Maeda, Yoshinobu Kimura

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   81 ( 7 )   1405 - 1408   2017年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    An effective method to prepare plant complex type (PCT) N-glycans in large amounts has been required to evaluate their immunological activity. In this study, we found that glycoproteins in bamboo shoots predominantly carry PCT N-glycans including the Lewis a epitope-containing ones, suggesting that bamboo shoot is an excellent source for the plant antigenic glycans to synthesize immunoactive neoglycopolymers.

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  • ヒトT細胞株への白石綿・青石綿継続曝露に伴う細胞特性の変化 査読

    李順姫, 前田恵, 松﨑秀紀, 武井直子, 山本祥子, 幡山圭代, 吉留 敬, 西村泰光, 大槻剛巳

    日本予防医学雑誌   10 ( 3 )   119 - 125   2016年12月

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    記述言語:日本語  

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  • 健康増進住居環境の構築:マイナス帯電粒子優位室内環境によるナチュラル・キラー活性の増強 査読

    西村泰光, 高橋一聡, 小谷宗男, 間瀬昭則, 白濱毅, 前田恵, 李順姫, 松崎秀紀, 武井直子, 吉留敬, 阿見和久, 大槻剛巳

    生存科学   26 ( 2 )   181 - 190   2016年10月

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    記述言語:日本語  

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  • 珪酸粒子・アスベスト繊維の免疫影響 査読

    大槻剛巳, 李順姫, 前田恵, 林宏明, 松﨑秀紀, 武井直子, 吉留敬, 西村泰光

    繊維状物質研究   3   4 - 11   2016年10月

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    記述言語:日本語   出版者・発行元:日本繊維状物質研究協会  

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  • FoxO1 regulates apoptosis induced by asbestos in the MT-2 human T-cell line. 査読 国際誌

    Matsuzaki H, Lee S, Maeda M, Kumagai-Takei N, Nishimura Y, Otsuki T

    Journal of immunotoxicology   13 ( 5 )   620 - 7   2016年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.3109/1547691X.2016.1143539

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  • Possible contribution of alpha2,6-sialylation to luteolysis in cows by inhibiting the luteotropic effects of galectin-1 査読 国際誌

    Kazuhisa Hashiba, Junko Nio-Kobayashi, Masahiro Sano, Megumi Maeda, Yoshinobu Kimura, Yuki Yamamoto, Koji Kimura, Kiyoshi Okuda

    Biology of Reproduction   95 ( 1 )   17 - 17   2016年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Society for the Study of Reproduction  

    The corpus luteum (CL) is essential for establishing pregnancy. If pregnancy does not occur during the estrous cycle, luteolysis is induced by prostaglandin (PG) F2alpha secreted from the uterus. Galectin-1, a beta-galactose-binding protein, is expressed in the functional CL of cows and increases the viability of bovine luteal steroidogenic cells (LSCs) by modifying the functions of membrane glycoproteins. The binding of galectin-1 to glycoproteins is blocked by alpha2,6-sialylation of the terminal galactose residues of glycoconjugates, which is catalyzed by a sialyltransferase (ST6Gal-I). However, the physiological role of alpha2,6-sialic acid in bovine CL is unclear. The level of alpha2,6-sialylation of the bovine CL was higher during the regressed-luteal stage than in other luteal stages. Lectin histochemistry revealed that alpha2,6-sialylated glycoconjugates were localized to luteal endothelial cells throughout the estrous cycle. In addition, alpha2,6-sialylated glycoconjugates concentrated to the membrane of LSCs during the regressed-luteal stage. PGF2alpha treatment for 72 h enhanced the expression of ST6Gal-I mRNA and the level of alpha2,6-sialylated glycoproteins in mid-LSCs. The level of alpha2,6-sialylated glycoproteins of late-stage LSCs (Days 15-17 after ovulation) was higher than that of mid-stage LSCs (Days 8-12 after ovulation), and galectin-1 increased the viability of mid-LSCs but not that of late-stage LSCs. Furthermore, galectin-1 increased the viability of late-LSCs when alpha2,6-sialic acid residues were removed by neuraminidase. The overall findings suggest that alpha2,6-sialylation stimulated by PGF2alpha contributes to luteolysis by inhibiting the luteotropic effects of galectin-1 in bovine CL.

    DOI: 10.1095/biolreprod.116.140194

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  • The proliferative effects of asbestos-exposed peripheral blood mononuclear cells on mesothelial cells. 査読 国際誌

    Yuho Maki, Yasumitsu Nishimura, Shinichi Toyooka, Junichi Soh, Kazunori Tsukuda, Kazuhiko Shien, Masashi Furukawa, Takayuki Muraoka, Tsuyoshi Ueno, Norimitsu Tanaka, Hiromasa Yamamoto, Hiroaki Asano, Megumi Maeda, Naoko Kumagai-Takei, Suni Lee, Hidenori Matsuzaki, Takemi Otsuki, Shinichiro Miyoshi

    Oncology letters   11 ( 5 )   3308 - 3316   2016年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Malignant mesothelioma (MM) is thought to arise from the direct effect of asbestos on mesothelial cells. However, MM takes a long time to develop following exposure to asbestos, which suggests that the effects of asbestos are complex. The present study examined the effects of asbestos exposure on the cell growth of MeT-5A human mesothelial cells via cytokines produced by immune cells. Peripheral blood mononuclear cells (PBMCs) were stimulated with antibodies against cluster of differentiation (CD)3 and CD28 upon exposure to the asbestos chrysotile A (CA) or crocidolite (CR); the growth of MeT-5A cells in media supplemented with PBMC culture supernatants was subsequently examined. MeT-5A cells exhibited an increase in proliferation when grown in supernatant from the 7-day PBMC culture exposed to CA or CR. Analysis of cytokine production demonstrated increased levels of granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-1α, IL-1β, IL-3, IL-5, IL-13 and IL-17A in supernatants. Individual administration of these cytokines, excluding G-CSF and GM-CSF, led to an increase in cell growth of MeT-5A, whereas this effect was not observed following the combined administration of these cytokines. The results indicate that cytokines secreted by immune cells upon exposure to asbestos cause an increase in the growth activity of mesothelial cells, suggesting that alterations in the production of cytokines by immune cells may contribute to tumorigenesis in individuals exposed to asbestos.

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  • Rice alpha-fucosidase active against plant complex type N-glycans containing Lewis a epitope: purification and characterization 査読 国際誌

    Md. Ziaur Rahman, Makoto Fujishige, Megumi Maeda, Yoshinobu Kimura

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   80 ( 2 )   291 - 294   2016年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    Rice -fucosidase (-fucosidase Os, 58kDa) that is active for 1-4 fucosyl linkage in Lewis a unit of plant N-glycans was purified to homogeneity. -fucosidase Os showed activity against 1-3 fucosyl linkage in Lacto-N-fucopentaose III but not 1-3 fucosyl linkage in the core of plant N-glycans. The N-terminal sequence of -fucosidase Os was identified as A-A-P-T-P-P-P-L-, and this sequence was found in the amino acid sequence of the putative rice -fucosidase 1 (Os04g0560400).

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  • Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope 査読 国際誌

    Megumi Maeda, Misato Tani, Takeo Yoshiie, Christopher J. Vavricka, Yoshinobu Kimura

    Carbohydrate Research   435   50 - 57   2016年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier Ltd  

    The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). The biological significance of Lewis a type plant N-glycans and their effects on the human immune system remain to be elucidated. Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. In this study, we have found that three water plants, Elodea nuttallii, Egeria densa, and Ceratophyllum demersum, produce glycoproteins bearing Lewis a units. Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts.

    DOI: 10.1016/j.carres.2016.09.008

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  • Enhancement of regulatory T cell-like suppressive function in MT-2 by long-term and low-dose exposure to asbestos. 査読 国際誌

    Ying C, Maeda M, Nishimura Y, Kumagai-Takei N, Hayashi H, Matsuzaki H, Lee S, Yoshitome K, Yamamoto S, Hatayama T, Otsuki T

    Toxicology   338   86 - 94   2015年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.tox.2015.10.005

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  • Exposure to negatively charged-particle dominant air-conditions on human lymphocytes in vitro activates immunological responses. 査読 国際誌

    Nishimura Y, Takahashi K, Mase A, Kotani M, Ami K, Maeda M, Shirahama T, Lee S, Matsuzaki H, Kumagai-Takei N, Yoshitome K, Otsuki T

    Immunobiology   220 ( 12 )   1359 - 68   2015年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • T cell alteration caused by exposure to asbestos 査読

    Megumi Maeda, Shoko Yamamoto, Tamayo Hatayama, Hidenori Matsuzaki, Suni Lee, Naoko Kumagai-Takei, Kei Yoshitome, Yasumitsu Nishimura, Yoshinobu Kimura, Takemi Otsuki

    Biological Effects of Fibrous and Particulate Substances   195 - 210   2015年10月

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    記述言語:英語   掲載種別:論文集(書籍)内論文   出版者・発行元:Springer Japan  

    A model to examine the effects of continuous exposure to asbestos on human T cells was established to interpret experimental findings for clinical utilization. Although transient exposure causes apoptosis in the human polyclonal cell line MT-2, continuous and relatively low-dose exposure resulted in resistance against asbestos-induced apoptosis with a higher production of TGF-β and IL-10 and subsequent resistance to TGF-β-induced growth inhibition and activation of STAT3 and Bcl-2. These alterations caused by continuous exposure to chrysotile asbestos were also observed in a subline exposed continuously to crocidolite and included resistance to apoptosis, changes of cytokine production, and demonstration of the importance of Bcl-2 for resistance against apoptosis. In addition, analysis of protein expression among the MT-2 original cell line, which was never exposed to asbestos, and the continuously exposed subline showed the phosphorylation of β-actin and the increasing level of cytoskeletal molecules. These findings indicate the importance of the cytoskeleton as the initial contact site between cells and asbestos fibers, particularly fibers that cannot move into the inside of cells because of their physical features. Finally, the CXCR3 chemokine receptor and related antitumor cytokine IFN-γ were assayed in these sublines continuously exposed to asbestos, as well as in vitro stimulated freshly isolated peripheral CD4 T cells derived from healthy donors and exposed to asbestos fibers. The CXCR3 expression and production capacity for IFN-γ were reduced by asbestos exposure, and these findings were also confirmed for peripheral CD4 T cells derived from patients with pleural plaque and malignant mesothelioma. The overall findings observed in continuously exposed human T cell models will contribute towards the early detection of asbestos exposure and occurrence of mesothelioma using peripheral blood and will improve the immune status (reducing antitumor immunity in asbestos-exposed patients) through the use of certain physiological substances derived from plants, foods, and microorganisms.

    DOI: 10.1007/978-4-431-55732-6_11

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  • Effects of asbestos fibers on human cytotoxic T cells 査読

    Naoko Kumagai-Takei, Yasumitsu Nishimura, Hidenori Matsuzaki, Megumi Maeda, Suni Lee, Kei Yoshitome, Takemi Otsuki

    Biological Effects of Fibrous and Particulate Substances   211 - 221   2015年10月

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    記述言語:英語   掲載種別:論文集(書籍)内論文   出版者・発行元:Springer Japan  

    The immunological effects of asbestos have been demonstrated and include reduction of antitumor immunity such as the reduction of natural killer (NK) cell activity with decreased expression of NK cell-activating receptor, NKp46, as well as reduced expression of CXCR3, chemokine receptor, and interferon (IFN)-γ in CD4+ T cells. In this review, the effects of asbestos as demonstrated by our investigations on the cellular characteristics and functions of cytotoxic T lymphocytes (CTLs) differentiated from CD8+ T cells are shown and discussed. Experiments involving in vitro exposure of chrysotile asbestos onto a mixed lymphocyte reaction (MLR) assay to estimate clonal expansion (proliferation and differentiation) of CD8+ T cells revealed the inhibition of both proliferation and differentiation. These results should be interpreted as the impairment of CTL differentiation in the lymph nodes, where asbestos fibers are located continuously in asbestos-exposed people. In addition, analyses of cellular functions in asbestos-exposed patients with pleural plaque (PP) and malignant mesothelioma (MM) are discussed. PP patients showed an increase in effector memory CD8+ T cells compared to healthy donors or MM patients. Furthermore, MM patients showed a decrease in perforin-expressing CD8+ T cells. These results indicated that although asbestos-exposed individuals are ready to respond with transformed cells, CTLs may lose their function once mesothelioma progresses.

    DOI: 10.1007/978-4-431-55732-6_12

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  • beta-Galactosidase from Ginkgo biloba seeds active against beta-galactose-containing N-glycans: purification and characterization 査読 国際誌

    Md. Ziaur Rahman, Megumi Maeda, Yoshinobu Kimura

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   79 ( 9 )   1464 - 1472   2015年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    In this study, we purified an acidic beta-galactosidase to homogeneity from Ginkgo biloba seeds (beta-Gal'ase Gb-1) with approximately 270-fold purification. A molecular mass of the purified beta-Gal'ase Gb-1 was estimated about 35kDa by gel filtration and 32kDa by SDS-PAGE under non-reducing condition, respectively. On the other hand, beta-Gal'ase Gb-1 produced a single band with a molecular mass of 16kDa by SDS-PAGE under reducing condition. The N-terminal amino acid sequences of 32kDa and 16kDa molecules were the same and identified as H-K-A-N-X-V-T-V-A-F-V-M-T-Q-H-, suggesting that beta-Gal'ase Gb-1 may function as a homodimeric structure in vivo. When complex-type N-glycans containing beta-galactosyl residues were used as substrates, beta-Gal'ase Gb-1 showed substantial activity for beta 1-4 galactosyl residue and modest activity for beta 1-3 galactosyl residue with an optimum pH near 5.0. Based on these results, the involvement of beta-Gal'ase Gb-1 in the degradation of plant complex-type N-glycans is discussed.

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  • Synthesis of a novel glycopolymer carrying multivalent plant antigenic N-glycans. 査読

    Maeda M, Takeda N, Kimura M, Kimura Y

    Glycoconjugate Journal   32 ( 5 )   193   2015年9月

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    記述言語:日本語  

    植物抗原性N-グリカンを多価数結合した新規糖鎖ポリマーの合成方法を確立した。

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  • Phenotype-based clustering of glycosylation-related genes by RNAi-mediated gene silencing 査読 国際誌

    Miki Yamamoto-Hino, Hideki Yoshida, Tomomi Ichimiya, Sho Sakamura, Megumi Maeda, Yoshinobu Kimura, Norihiko Sasaki, Kiyoko F. Aoki-Kinoshita, Akiko Kinoshita-Toyoda, Hidenao Toyoda, Ryu Ueda, Shoko Nishihara, Satoshi Goto

    Genes to Cells   20 ( 6 )   521 - 542   2015年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Blackwell Publishing Ltd  

    Glycan structures are synthesized by a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters. For example, the common core region of glycosaminoglycans (GAGs) is sequentially synthesized by peptide-O-xylosyltransferase, β1,4-galactosyltransferase I, β1,3-galactosyltransferase II, and β1,3-glucuronyltransferase. This raises the possibility that functional impairment of GR proteins involved in synthesis of the same glycan might result in the same phenotypic abnormality. To examine this possibility, comprehensive silencing of genes encoding GR and proteoglycan core proteins was conducted in Drosophila. Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. Spatiotemporally regulated silencing caused a range of malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation. The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). Based on the phenotypic clustering, CG33145 was predicted to be involved in formation of terminal domains. Our further analysis showed that CG33145 exhibited galactosyltransferase activity in synthesis of terminal N-linked glycans. Phenotypic clustering, therefore, has potential for the functional prediction of novel GR genes.

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  • Identification of β1,3-galactosyltransferases responsible for biosynthesis of insect complex-type N-glycans containing a T-antigen unit in the honeybee 査読 国際誌

    Tomomi Ichimiya, Megumi Maeda, Shou Sakamura, Masato Kanazawa, Shoko Nishihara, Yoshinobu Kimura

    Glycoconjugate Journal   32 ( 3-4 )   141 - 151   2015年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Kluwer Academic Publishers  

    Honeybees (Apis mellifera) produce unique complex-type N-glycans bearing a Galβ1-3GalNAc (T-antigen) unit, and honeybee-specific N-glycans are linked to royal jelly glycoproteins. In this study, we identified two novel honeybee β1,3-galactosyltransferase (β1,3-GalT) genes responsible for biosynthesis of the T-antigen in insect N-glycans. The products of the two putative β1,3-GalT genes (β1,3-GalT1 and β1,3-GalT2), which were expressed in Sf21 insect cells, transferred galactose (Gal) residues to GalNAc2GlcNAc2Man3GlcNAc2-PA to form the Galβ1-3GalNAc unit, indicating that the identified genes were involved in biosynthesis of the β1-3 Gal-containing N-glycan. Therefore, using biochemistry and molecular biology techniques, we revealed a unique N-glycan biosynthesis mechanism in the cephalic region of honeybees, which has not previously been found in other animal or plant cells.

    DOI: 10.1007/s10719-015-9585-7

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  • 抗腫瘍免疫への石綿曝露影響 査読

    武井直子, 西村泰光, 前田恵, 林宏明, 松崎秀紀, 李順姫, 吉留敬, 岸本卓巳, 福岡和也, 中野孝司, 大槻剛巳

    日本予防医学会雑誌   10 ( 1 )   1 - 8   2015年4月

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  • Enhancement of NK Cell Cytotoxicity Induced by Long-Term Living in Negatively Charged-Particle Dominant Indoor Air-Conditions. 査読 国際誌

    Nishimura Y, Takahashi K, Mase A, Kotani M, Ami K, Maeda M, Shirahama T, Lee S, Matsuzaki H, Kumagai-Takei N, Yoshitome K, Otsuki T

    PloS one   10 ( 7 )   e0132373   2015年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1371/journal.pone.0132373

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  • Functional Alteration of Natural Killer Cells and Cytotoxic T Lymphocytes upon Asbestos Exposure and in Malignant Mesothelioma Patients. 査読 国際誌

    Nishimura Y, Kumagai-Takei N, Matsuzaki H, Lee S, Maeda M, Kishimoto T, Fukuoka K, Nakano T, Otsuki T

    BioMed research international   2015   238431 - 238431   2015年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1155/2015/238431

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  • Cloning and expression analysis of acidic peptide:N-glycanase genes in Solanum lycoperisicum. 査読

    Fujikawa, M, Maeda, M, Fujiyama , K, Kimura, Y

    Glycoconjugate Journal   32 ( 5 )   193   2015年

  • Purification and characterization of Ginkgo biloba α-1,3/4-fucosidase active against Lewis a antigen 査読

    Itano, S, Maeda, M, Kimura, Y

    Glycoconjugate Journal   32 ( 5 )   195   2015年

  • Chronic exposure to asbestos enhances TGF-β1 production in the human adult T cell leukemia virus-immortalized T cell line MT-2. 査読 国際誌

    Maeda M, Chen Y, Hayashi H, Kumagai-Takei N, Matsuzaki H, Lee S, Nishimura Y, Otsuki T

    International journal of oncology   45 ( 6 )   2522 - 32   2014年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.3892/ijo.2014.2682

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  • 【レギュラトリーT細胞の機能発現】 制御性T細胞機能へのアスベスト曝露影響 査読

    西村 泰光, 三浦 由恵, 前田 恵, 松崎 秀 紀, 李 順姫, 武井 直子, 大槻 剛巳

    剛巳【レギュラトリーT細胞の機能発現】 制御性T細胞機能へのアスベスト曝露影響 (解説/特集) 臨床免疫・アレルギー科   62 ( 4 )   366 - 370   2014年10月

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    記述言語:日本語  

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  • Silica exposure and altered regulation of autoimmunity. 査読 国際誌

    Lee S, Matsuzaki H, Kumagai-Takei N, Yoshitome K, Maeda M, Chen Y, Kusaka M, Urakami K, Hayashi H, Fujimoto W, Nishimura Y, Otsuki T

    Environmental health and preventive medicine   19 ( 5 )   322 - 9   2014年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1007/s12199-014-0403-9

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  • アスベスト曝露によるTh1細胞機能の低下 査読

    西村 泰光, 前田 恵, 三浦 由恵, 松崎 秀 樹, 李 順姫, 武井 直子, 大槻 剛巳

    .臨床免疫・アレルギー科   62 ( 3 )   314 - 317   2014年9月

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    記述言語:日本語   出版者・発行元:科学評論社  

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    その他リンク: http://search.jamas.or.jp/link/ui/2015007761

  • 珪酸曝露による免疫影響 査読

    松﨑秀紀, 前田恵, 林宏明, 武井直子, 李順姫, 吉留敬, 草加勝康, 浦上更三, 西村泰光, 兵藤文則, 大槻剛巳

    日本予防医学会雑誌   9 ( 1 )   1 - 8   2014年4月

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  • Large-scale preparation of glycopeptides harboring the TF-antigen unit from royal jelly 査読 国際誌

    Megumi Maeda, Tatsuya Tanaka, Mariko Kimura, Yoshinobu Kimura

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   78 ( 2 )   276 - 278   2014年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    We have reported that new N-glycans carrying the TF-antigen occurred on a major royal jelly glycoprotein, and we have identified the glycosylation site to which the antigenic N-glycan is linked, but an appropriate procedure has not been established to prepare non-labeled immunoreactive glycopeptides in large amounts for functional analysis. In this study, we developed an effective method of preparing Asn-glycopeptide bearing TF-antigen.

    DOI: 10.1080/09168451.2014.878223

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  • Functional properties of CD8(+) lymphocytes in patients with pleural plaque and malignant mesothelioma. 査読 国際誌

    Kumagai-Takei N, Nishimura Y, au, Maeda M, Hayashi H, Matsuzaki H, Lee S, Kishimoto T, Fukuoka K, Nakano T, Otsuki T

    Journal of immunology research   2014   670140 - 670140   2014年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1155/2014/670140

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  • 珪肺症例における自己寛容破綻の異常

    大槻 剛巳, 前田 恵, 松崎 秀紀, 李 順姫, 武井 直子, 西村 泰光

    日本毒性学会学術年会   41.1   O-39   2014年

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    記述言語:日本語   出版者・発行元:日本毒性学会  

    珪酸とアスベストの慢性職業性曝露は塵肺症として知られる肺の繊維化を惹起する。珪酸曝露症例は珪肺症と呼ばれるが,合併症として強皮症,関節リウマチ(Caplan症候群として知られる),ANCA関連血管炎などの自己免疫疾患が知られている。従来,珪酸のアジュバントとしての作用と捉えられてきたが,我々は,珪酸曝露が直接免疫系に対して,なんらかの影響(毒性あるいは活性化)を惹起する可能性を考えて,実験系あるいは症例の末梢血単核球や,リンパ球などを用いて検討を加えてきた。アスベストについては,悪性中皮腫や肺癌などの悪性腫瘍の合併が知られており,この観点でもアスベストによる免疫系への毒性としての,腫瘍免疫の減衰について,いくつかの知見を得ているが,本演題では珪酸の免疫担当細胞への影響について,報告する。まずリンパ球の細胞死に関連が深いCD95/Fas分子について,可溶性Fasやその他のFas媒介アポトーシス阻害因子が珪肺症例で上昇していること。一方,機能性抗Fas自己抗体の検出や内因性生理的Fas誘導アポトーシスの阻害因子の珪肺症例末梢血単核球での発現減弱などの所見より,珪肺症例では慢性活性化を受け,長期生存にいたり,おそらくは自己抗原とも反応している一群と,細胞死に陥ってはリクルートされている一群がある可能性が想起された。前者は,反応性T細胞で,その他可溶性IL-2受容体の血清中での上昇や,Decoy Receptor 3 (DcR3)分子の血漿中濃度上昇,PD-1 mRNAの発現上昇などの活性化指標が検出された。一方,後者は制御性T細胞の可能性があり,珪肺症例制御性T細胞では活性化に伴うFas分子の過剰発現とともに,早期の細胞死が誘導されることが観察された。これらの免疫バランスの異常が,珪肺症例での自己寛容の破綻を惹起している可能性がある。

    DOI: 10.14869/toxpt.41.1.0_o-39

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  • Alteration of cytoskeletal molecules in a human T cell line caused by continuous exposure to chrysotile asbestos. 査読 国際誌

    Maeda M, Chen Y, Kumagai-Takei N, Hayashi H, Matsuzaki H, Lee S, Hiratsuka J, Nishimura Y, Kimura Y, Otsuki T

    Immunobiology   218 ( 9 )   1184 - 91   2013年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.imbio.2013.04.007

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  • Purification and Substrate Specificity of A Ginkgo biloba Glycosidase Active in beta-1,2-Xylosidic Linkage in Plant Complex Type N-Glycans 査読 国際誌

    Megumi Maeda, Tsuyoshi Akiyama, Daisuke Yokouchi, Kwan Kit Woo, Yoshinobu Kimura

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   77 ( 9 )   1973 - 1976   2013年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    The beta-xylosidase, which is active against plant complex type N-glycans, was purified to homogeneity from Ginkgo biloba seeds. The N-terminal amino acid sequence, G-S-A-A-G-N-R-, of the Ginkgo beta-xylosidase (beta-Xyl'ase Gb) was consistent with the deduced internal amino acid sequence of an Arabidopsis beta-xylosidase (AtBXL1). beta-Xyl'ase Gb hydrolyzed the beta 1-2 xylosyl residue from Xyl beta 1-2Man beta 1-4G1cNAc beta 1-4GlcNAc-PA and Xyl beta 1-2Man beta 1-4G1cNA beta 1-4(Fuc alpha 1-3)GlcNAc-PA, but not that from Manal-6(Man alpha 1-3)(Xy1 beta 1-2)Man beta 1-4GlcNAc beta 1-4(Fuc alpha 1-3)GlcNAc-PA.

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  • Effect of asbestos exposure on differentiation of cytotoxic T lymphocytes in mixed lymphocyte reaction of human peripheral blood mononuclear cells. 査読 国際誌

    Kumagai-Takei N, Nishimura Y, au, Maeda M, Hayashi H, Matsuzaki H, Lee S, Hiratsuka J, Otsuki T

    American journal of respiratory cell and molecular biology   49 ( 1 )   28 - 36   2013年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1165/rcmb.2012-0134OC

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  • Purification and characterization of beta-xylosidase that is active for plant complex type N-glycans from tomato (Solanum lycopersicum): removal of core alpha 1-3 mannosyl residue is prerequisite for hydrolysis of beta 1-2 xylosyl residue 査読 国際誌

    Daisuke Yokouchi, Natsuko Ono, Kosuke Nakamura, Megumi Maeda, Yoshinobu Kimura

    GLYCOCONJUGATE JOURNAL   30 ( 5 )   463 - 472   2013年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER  

    In this study, we purified and characterized the beta-xylosidase involved in the turnover of plant complex type N-glycans to homogeneity from mature red tomatoes. Purified beta-xylosidase (beta-Xyl'ase Le-1) gave a single band with molecular masses of 67 kDa on SDS-PAGE under a reducing condition and 60 kDa on gelfiltration, indicating that beta-Xyl'ase Le-1 has a monomeric structure in plant cells. The N-terminal amino acid could not be identified owing to a chemical modification. When pyridylaminated (PA-) N-glycans were used as substrates, beta-Xyl'ase Le-1 showed optimum activity at about pH 5 at 40 A degrees C, suggesting that the enzyme functions in a rather acidic circumstance such as in the vacuole or cell wall. beta-Xyl'ase Le-1 hydrolyzed the beta 1-2 xylosyl residue from Man(1)Xyl(1)GlcNAc(2)-PA, Man(1)Xyl(1)Fuc(1)GlcNAc(2)-PA, and Man(2)Xyl(1)Fuc(1)GlcNAc(2)-PA, but not that from Man(3)Xyl(1)GlcNAc(2)-PA or Man(3)Xyl(1)Fuc(1)GlcNAc(2)-PA, indicating that the alpha 1-3 arm mannosyl residue exerts significant steric hindrance for the access of beta-Xyl'ase Le-1 to the xylosyl residue, whereas the alpha 1-3 fucosyl residue exerts little effect. These results suggest that the release of the beta 1-2 xylosyl residue by beta-Xyl'ase Le-1 occurs at least after the removal the alpha-1,3-mannosyl residue in the core trimannosyl unit.

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  • Altered functions of alveolar macrophages and NK cells involved in asbestos-related diseases. 査読 国際誌

    Nishimura Y, Maeda M, Kumagai-Takei N, Lee S, Matsuzaki H, Wada Y, Nishiike-Wada T, Iguchi H, Otsuki T

    Environmental health and preventive medicine   18 ( 3 )   198 - 204   2013年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1007/s12199-013-0333-y

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  • Exploration of biomarkers for asbestos exposure and occurrence of malignant mesothelioma based on the immunological effects of asbestos.

    Matsuzaki H, Lee S, Kumagai-Takei N, Hayashi H, Miura Y, Chen Y, Maeda M, Yamamoto S, Hatayama T, Nishimura Y, Otsuki T

    Journal of Data Mining Genomics Proteomics   S2   001   2013年

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  • アスベストの免疫毒性学的側面と病態への関与

    大槻 剛巳, 前田 恵, 武井 直子, 松崎 秀紀, 李 順姫, 西村 泰光

    日本毒性学会学術年会   40.1   1114   2013年

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    記述言語:日本語   出版者・発行元:日本毒性学会  

    アスベストによる発癌性については,含有する鉄による酸化ストレス,繊維状物質としての物理的なDNA損傷,そしてその他の発癌物質の吸着などが考えられる。一方,アスベストは珪酸(SiO2)の金属塩であり,珪酸曝露症例(珪肺症)にて自己免疫疾患が多く合併することや,その実験的検証から,アスベストもまた免疫毒性あるいは免疫担当細胞への作用が想定される。さらにアスベスト曝露者の合併症として悪性中皮腫などの発癌,初期曝露からの長期の潜伏期を考えると,抗腫瘍免疫の減衰などを惹起している可能性がある。我々は,アスベストの免疫担当細胞への影響ならびに抗腫瘍免疫との関連について,検討を加えている。細胞株モデル,健常人末梢血から採取した新鮮免疫担当細胞,あるいは曝露者(胸膜プラークや悪性中皮腫症例)の末梢血のリンパ球の解析を行ってきている。T細胞では,細胞株や,健常人由来細胞にて長期曝露によってアスベストが誘導するアポトーシスに抵抗性を獲得することが判明し,その機序として,Src-kinase,IL-10,STAT3,BCL-2の関与が判明した。さらにこれらの細胞では,抗腫瘍免疫にも重要なCXCR3発現やIFNγ産生の低下も認められた。制御性T細胞では細胞株モデルにて可用性因子であるIL-10,TGFβの産生過剰とともに細胞間接着による反応性T細胞の活性化抑制能の増強も確認された。制御性T細胞ではマスタージーンであるFoxP3を制御するFOXO1の発現低下と,それに伴う細胞周期の異調節なども観察されている。また,CD8+細胞障害性T細胞においても,実験系での分化誘導・増殖時のアスベスト曝露での抑制が判明した。一方,NK細胞においても,細胞内シグナリングの減弱とともに,機能発現活性化受容体の発現減弱が認められ, 特にNKp46分子については,アスベスト曝露症例においても,その殺細胞能との相関が認められた。さらなる検討とともに,曝露や担癌の指標あるいは予防的観点からのこれらの成果の応用も期待される。

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  • Structural features of N-glycans of seaweed glycoproteins: predominant occurrence of high-mannose type N-glycans in marine plants. 査読 国際誌

    Yoshiie T, Maeda M, Kimura M, Hama Y, Uchida M, Kimura Y

    Bioscience, biotechnology, and biochemistry   76 ( 10 )   1996 - 1998   2012年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1271/bbb.120463

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  • Resistance to asbestos-induced apoptosis with continuous exposure to crocidolite on a human T cell. 査読 国際誌

    Maeda M, Yamamoto S, Chen Y, Kumagai-Takei N, Hayashi H, Matsuzaki H, Lee S, Hatayama T, Miyahara N, Katoh M, Hiratsuka J, Nishimura Y, Otsuki T

    The Science of the total environment   429   174 - 82   2012年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.scitotenv.2012.04.043

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  • Environmental factors producing autoimmune dysregulation--chronic activation of T cells caused by silica exposure. 査読 国際誌

    Lee S, Hayashi H, Maeda M, Chen Y, Matsuzaki H, Takei-Kumagai N, Nishimura Y, Fujimoto W, Otsuki T

    Immunobiology   217 ( 7 )   743 - 8   2012年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.imbio.2011.12.009

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  • Asbestos-induced cellular and molecular alteration of immunocompetent cells and their relationship with chronic inflammation and carcinogenesis. 査読 国際誌

    Matsuzaki H, Maeda M, Lee S, Nishimura Y, Kumagai-Takei N, Hayashi H, Yamamoto S, Hatayama T, Kojima Y, Tabata R, Kishimoto T, Hiratsuka J, Otsuki T

    Journal of biomedicine & biotechnology   2012   492608 - 492608   2012年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1155/2012/492608

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  • Review of reduced tumor immunity caused by asbestos exposure to immunocompetent cells such as T and NK cells. 査読

    Otsuki T, Lee S, Kumagai-Takei N, Miyahara N, Katoh M, Yamamoto S, Hatayama T, Matsuzaki H, Nishimura Y, Maeda M, Hayashi H, Hirastuka J, Chen Y

    Open Access Scientific reports   1 ( 8 )   open access   2012年

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  • Decreased CXCR3 expression in CD4+ T cells exposed to asbestos or derived from asbestos-exposed patients. 査読 国際誌

    Megumi Maeda, Yasumitsu Nishimura, Hiroaki Hayashi, Naoko Kumagai, Ying Chen, Shuko Murakami, Yoshie Miura, Jun-ichi Hiratsuka, Takumi Kishimoto, Takemi Otsuki

    American journal of respiratory cell and molecular biology   45 ( 4 )   795 - 803   2011年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Asbestos causes malignant tumors such as lung cancer and malignant mesothelioma (MM). To determine whether asbestos exposure causes reduction of antitumor immunity, we established an in vitro T-cell line model of low-dose and continuous exposure to asbestos using an human adult T-cell leukemia virus-1 immortalized human polyclonal T-cell line, MT-2, and revealed that MT-2 cells exposed continuously to asbestos showed resistance to asbestos-induced apoptosis. In addition, the cells presented reduction of surface CXCR3 chemokine receptor expression and IFN-γ production. In this study, to confirm that these findings are suitable for clinical translation, surface CXCR3 and IFN-γ expression were analyzed using freshly isolated human CD4(+) T cells derived from healthy donors and patients with pleural plaque (PP) or MM. The results revealed that CXCR3 and IFN-γ expression in the ex vivo model were reduced in some cases. Additionally, CXCR3 expression in CD4(+) T cells from PPs and MMs was significantly reduced compared with that from healthy donors, and CD4(+) T cells from patients with MMs exhibited a marked reduction in IFN-γ mRNA levels after stimulation in vitro. Moreover, CD4(+)CXCR3(+) T cells in lymphocytes from MMs showed a tendency for an inverse correlation with its ligand CXCL10/IP10 in plasma. These findings show reduction of antitumor immune function in asbestos-exposed patients and indicate that CXCR3, IFN-γ, and CXCL10/IP10 may be candidates to detect and monitor disease status.

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  • 川崎医科大学における大学連携、産学官連携等、対外活動について:その2

    大槻剛巳, 小笠原康夫, 柏原直樹, 佐藤稔, 大澤裕, 矢田豊隆, 毛利聡, 山内明, 武井直子, 前田恵, 西村泰光, 小野寺昇, 望月精一, 茅野功, 川西礼美, 福永仁夫

    川崎医学会誌   37   47 - 59   2011年9月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:川崎医科大学  

    DOI: 10.11482/KMJ-LAS(37)47-59.2011

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  • Reduction of CXC chemokine receptor 3 in an in vitro model of continuous exposure to asbestos in a human T-cell line, MT-2. 査読 国際誌

    Megumi Maeda, Yasumitsu Nishimura, Hiroaki Hayashi, Naoko Kumagai, Ying Chen, Shuko Murakami, Yoshie Miura, Jun-ichi Hiratsuka, Takumi Kishimoto, Takemi Otsuki

    American journal of respiratory cell and molecular biology   45 ( 3 )   470 - 9   2011年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Because patients with silicosis who are chronically exposed to silica particles develop not only pulmonary fibrosis, but also complications involving autoimmune diseases such as rheumatoid arthritis and systemic sclerosis, exposure to asbestos may affect the human immune system. This immunologic effect may impair antitumor immune function because cancer complications such as lung cancer and malignant mesothelioma are found in patients exposed to asbestos. To elucidate the antitumor immune status caused by CD4(+) T cells exposed to asbestos, an in vitro T-cell model of long-term and low-level exposure to chrysotile asbestos was established from a human adult T-cell leukemia virus-1-immortalized human polyclonal T cell line, MT-2, and the resulting six sublines showed resistance to asbestos-induced apoptosis after more than 8 months of continuous exposure. The results of DNA microarray analysis showed that the expression of 139 genes was altered by long-term and low-level exposure to asbestos, and the profile was almost similar among the six sublines when compared with the original MT-2 cells that had never been exposed to asbestos. Pathway and network analysis indicated a down-regulation of IFN-γ signaling and expression of CXC chemokine receptor 3 (CXCR3) in the sublines, whereas ELISA and flow cytometry analysis demonstrated a reduction in Th1-related IFN-γ production and cell-surface CXCR3 expression. These findings suggest that chronic exposure to asbestos may reduce antitumor immune status in CD4(+) T cells, and that an in vitro T-cell model may be useful in identifying molecules related to the impairment of antitumor immune function.

    DOI: 10.1165/rcmb.2010-0213OC

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  • Effects of environmental substance on the human immune system - Reduction of tumor immunity induced by asbestos exposure

    Naoko Kumagai, Megumi Maeda, Hidenori Matsuzaki, Suni Lee, Yasumitsu Nishimura, Takemi Otsuki

    Biotherapy   25 ( 4 )   739 - 746   2011年7月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)  

    Similar to the silica which causes dysregulation of autoimmunity as well as pulmonary fibrosis, asbestos, which is mineral silicate including iron, magnesium or calcium attached to the core silica molecule, SiO2 may affect the human immune system. Considering malignant complications of asbestos-exposed patients such as mesothelioma and lung cancer, asbestos is assumed to lower tumor immunity. The effects of asbestos on tumor immunity have been investigated with this in mind. It was found that natural killer (NK) cell activity was reduced in correspondence with the surface expression level of NKp46, one of the activating receptors of the NK cell. The proliferation and differentiation of cytotoxic T lymphocytes (CTL) were inhibited during induction of effector/memory CTL from naïve CD8+ T cells by culturing with allo-antigen supplemented with chrysotile-asbestos. In addition, tumor immunity-related molecules such as interferon (IFN) -γ and the CXCR3 chemokine receptor, were reduced by asbestos exposure using the T cell line, MT-2. Furthermore, activation-inhibitory functions such as the CD4 +CD25+FoxP3+ regulatory T cell possessing the MT-2 cell line, was enhanced by asbestos exposure. All of these findings suggested that asbestos exposure lowered tumor immunity. In addition, some of these findings were confirmed in asbestos-exposed patients. Reduction of tumor immunity caused by exposure to asbestos may make it easier to develop asbestos-inducing malignant tumors in the patients with asbestos exposure.

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  • Quick detection of overexpressed genes caused by myeloma-specific chromosomal translocations using multiplex RT-PCR. 査読 国際誌

    San San Htwe, Megumi Maeda, Rui Matsumoto, Natsumi Sakamoto, Shuko Murakami, Shoko Yamamoto, Minako Katoh, Naoko Kumagai, Hiroaki Hayashi, Yasumitsu Nishimura, Mitsugi Ohkura, Hideho Wada, Masafumi Taniwaki, Takashi Sugihara, Takemi Otsuki

    International journal of molecular medicine   27 ( 6 )   789 - 94   2011年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Multiple myeloma (MM) is a malignancy of the plasmocyte and is associated with various symptoms such as anemia, immunodeficiency, bone lesions and kidney insufficiency. Although prognosis was poor until some years ago, recent advances that introduced newer molecular targeting agents such as bortezomib and thalidomide have resulted in a better prognosis for MM. However, clinical manifestations and the relationship between cellular and molecular findings, including chromosomal translocation and the related overexpression of oncogenes such as CCND1 (cyclin D1) and FGFR3 (fibroblast growth factor receptor 3), remain unclear. It has been reported that a specific translocation may influence the prognosis of MM. Although translocations and overexpressed genes should be examined in ordinary clinical investigations, limited definitive assays for translocation involve the use of FISH (fluorescent in situ hybridization) or SKY (special karyotypic) methods. We therefore, attempted to establish a quick detection method for major translocated genes such as FGFR3, CCND1, CCND3 and MAF using multiplex RT-PCR (MP-RT-PCR). MP-RT-PCR can be performed within several to 24 h after bone marrow samples are taken. Two of 21 bone marrow blood samples from MM patients were analyzed using MP-RT-PCR and double-color FISH, and the results of both methods were compatible. Future utilization and elaboration of this method may help our understanding of the cell biology and clinical features of MM.

    DOI: 10.3892/ijmm.2011.648

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  • 珪酸の免疫影響と自己寛容の破綻 査読

    李順姫, 林宏明, 前田恵, 熊谷直子, 松﨑秀紀, 草加勝康, 浦上更三, 藤本亘, 大槻剛巳

    臨床環境医学   20   40 - 53   2011年6月

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  • Double-Knockout of Putative Endo-beta-N-acetylglucosaminidase (ENGase) Genes in Arabidopsis thaliana: Loss of ENGase Activity Induced Accumulation of High-Mannose Type Free N-Glycans Bearing N,N '-Acetylchitobiosyl Unit 査読 国際誌

    Yoshinobu Kimura, Yuki Takeoka, Masami Inoue, Megumi Maeda, Kazuhito Fujiyama

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   75 ( 5 )   1019 - 1021   2011年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    Endo-beta-N-acetylglucosaminidase (ENGase) is involved in the production of high-mannose type free N-glycans during plant development and fruit maturation. In a previous study (K. Nakamura et al. Biosci. Biotechnol. Biochem., 73, 461-464 (2009)), we identified the tomato ENGase gene and found that gene expression remained relatively constant. In the present study, we constructed an Arabidopsis thaliana mutant in which the expression of two putative ENGase genes was suppressed. The mutant showed no ENGase activity, but produced high-mannose type free N-glycans carrying the N,N'-acetylchitobiosyl unit that is produced by peptide:N-glycanase, indicating that both these genes encode Arabidopsis ENGase.

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  • ヒト免疫担当細胞への環境中物質の影響―アスベスト曝露による腫瘍免疫減衰― 査読

    熊谷直子, 前田恵, 松﨑秀紀, 李順姫, 西村泰光, 大槻剛巳

    Biotherapy   25 ( 4 )   739 - 746   2011年4月

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  • Dysregulation of autoimmunity caused by silica exposure and alteration of Fas-mediated apoptosis in T lymphocytes derived from silicosis patients 査読

    Otsuki T, Hayashi H, Nishimura Y, Hyodo F, Maeda M, Kumagai N, Miura Y, Kusaka M, Urakami K

    Int J Immunopath Pharmacol   24 ( 1S )   11S - 16S   2011年1月

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    記述言語:英語  

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  • Suppressive effect of asbestos on cytotoxicity of human NK cells 査読

    Nishimura Y, Kumagai N, Maeda M, Hayashi H, Fukuoka K, Nakano T, Miura Y, Hiratsuka J, Otsuki T

    Int J Immunopath Pharmacol   24 ( 1S )   5S - 10S   2011年1月

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  • Asbestos induces reduction of tumor immunity. 査読 国際誌

    Kumagai-Takei N, Maeda M, Chen Y, Matsuzaki H, Lee S, Nishimura Y, Hiratsuka J, Otsuki T

    Clinical & developmental immunology   2011   481439 - 481439   2011年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1155/2011/481439

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  • Structural features and Immunological activity of N-glycans of seaweed glyco- proteins. 査読

    Yoshiie T, Maeda M, Kimura M, Kimura Y

    Glycoconjugate Journal   28 ( 5 )   2011年

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    ホンダワラなど数種の海藻に含まれるN-グリカンの構造を明らかにするとともに、それらのヒト免疫細胞に対する免疫賦活化作用について解析した。

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  • Establishment of negatively-charged indoor air conditions and their biological effects

    Takemi Otsuki, Kazuaki Takahashi, Akinori Mase, Takashi Kawado, Muneo Kotani, Yasumitsu Nishimura, Megumi Maeda, Shuko Murakami, Naoko Kumagai, Hiroaki Hayashi, Ying Chen, Yoshie Miura, Takashi Shirahama, Michiharu Yoshimatsu, Kanehisa Morimoto

    Buildings and the Environment   201 - 214   2010年12月

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    Indoor air condition may be involved to the health status. Among various factors inindoor air conditions, few reports were published concerning negatively chargedparticles.In this review, we detaile the biological effects of negatively-charged indoorair conditions, particularly on the psycho-neuro-endocrino-immune network. The short-term (2.5 hours) and mmedium-term (2 weeks) abidance of negatively-charged airparticle dominant room resulted activation of natural killer cell activity induced byrecurrent transient activation of interleukin 2. These results indicate that negativelychargedindoor air condition is better for immune status and may improve daily lives. © 2009 Nova Science Publishers, Inc. All rights reserved.

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  • 珪酸・アスベストの免疫影響 (ミニ特集:繊維・粒子状物質研究会(仙台)) 査読

    熊谷直子, 西村泰光, 前田 恵, 林 宏明, 大槻剛巳

    日本衛生学雑誌   64 ( 4 )   493 - 499   2010年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • Dysregulation of the immune system caused by silica and asbestos. 査読

    Maeda M, Nishimura Y, Kumagai N, Hayashi H, Hatayama T, Katoh M, Miyahara N, Yamamoto S, Hirastuka J, Otsuki T

    Journal of immunotoxicology   7 ( 4 )   268 - 278   2010年10月

  • [Immunological effects of silica/asbestos]. 査読

    Kumagai N, Nishimura Y, Maeda M, Hayashi H, Otsuki T

    Nihon eiseigaku zasshi. Japanese journal of hygiene   65 ( 4 )   493 - 9   2010年9月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1265/jjh.65.493

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  • Intracellular and extracellular free N-glycans produced by plant cells: Occurrence of unusual plant complex-type free N-glycans in extracellular speces. 査読 国際誌

    Maeda M, Kimura M, Kimura Y

    J. Biocem.   148 ( 6 )   681 - 692   2010年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    植物細胞の新生たんぱく質の品質管理系における脱グリコシル化機構を明らかにする目的で、イネ培養細胞が産生する遊離型N-グリカンの構造特性解析を行った。得られた結果は、これまでに知られていない新たな糖鎖遊離機構が存在することを示唆するものであった。

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  • 珪肺症例における自己免疫異常の検出 血清可溶性インターロイキン2受容体と可溶性CD40リガンドの濃度の意義

    林 宏明, 西村 泰光, 前田 恵, 熊谷 直子, 村上 周子, 三浦 由恵, 草加 勝康, 浦上 更三, 大槻 剛巳

    日本職業・災害医学会会誌   58 ( 2 )   45 - 51   2010年3月

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    記述言語:日本語   出版者・発行元:(一社)日本職業・災害医学会  

    珪肺症例では肺病変のみならず自己寛容の破綻が生じ、慢性関節リウマチや強皮症の合併が知られている。これらの自己免疫異常の血清学的な指標として可溶性インターロイキン2受容体(sIL-2R)や可溶性CD40リガンド(sCD40L)が有用かどうかを健常人、珪肺症例、強皮症症例で検討した結果を総括して、珪酸の制御性T細胞や活性化T細胞など、自己寛容の破綻に関連する細胞群への影響について概説する。(著者抄録)

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  • Reductive alteration of the regulatory function of the CD4(+)CD25(+) T cell fraction in silicosis patients. 査読

    H. Hayashi, Y. Miura, M. Maeda, S. Murakami, N. Kumagai, Y. Nishimura, M. Kusaka, K. Urakami, W. Fujimoto, T. Otsuki

    International journal of immunopathology and pharmacology   23   1099 - 1109   2010年1月

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    Causal links have been documented between silica and rheumatoid arthritis, lupus erythematosus, systemic sclerosis and glomerulonephritis. Two different effects of silica have been suggested, an enhanced inflammatory response in the pulmonary region (e.g. activation of alveolar macrophages) and dysregulation of autoimmunity. Based on our previous reports showing in vitro activation of peripheral T cells by silica and reduced regulatory function of the peripheral CD4(+)CD25(+) fraction in which FoxP(3)+ regulatory T cells (Treg) are located, reconstitution of the CD4(+)CD25(+) fraction in silicosis patients (SILs) was investigated. Since T cells in peripheral CD4(+)CD25(+) and CD4(+)CD25(-) (effector T cells; Teff) fractions from SILs showed higher expression of pd-1 (a marker gene for T cell activation) in comparison to that of healthy donors (HDs), chronic T cell activation was considered to have occurred in SILs. In this study, a higher expression of the CD95/Fas molecule in Treg was recorded from silicosis patients (SILs) compared to healthy donors (HDs), and excess loss of FoxP3(+) Treg in freshly isolated peripheral blood mononuclear cells (PBMCs) from SILs relative to HDs was demonstrated when these cells were cultured with silica ex vivo, whereas CD25(+) cells were not reduced due to contamination of activated Teff in the CD4(+)CD25(+) fraction. The activation of both Teff and Treg results in reconstitution of the peripheral CD4(+)CD25(+) fraction, loss of Treg and contamination of activated Teff, resulting in reduction of the number and function of Treg. These results contribute to our understanding of the development of autoimmune diseases found in SILs.

    DOI: 10.1177/039463201002300414

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  • Is soluble CD40 ligand an indicator of immunopathological disturbance in silicosis patients? 査読

    HAYASHI Hiroaki, NISHIMURA Yasumitsu, MAEDA Megumi, MURAKAMI Shuko, KUMAGAI Naoko, CHEN Ying, KUSAKA Masayasu, URAKAMI Kozo, FUJIMOTO Wataru, OTSUKI Takemi

    川崎医学会誌   35 ( 2 )   129 - 138   2009年12月

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    記述言語:英語   出版者・発行元:川崎医科大学  

    DOI: 10.11482/2009/35.129.2009_KMJ_Hayashi_etal.pdf

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  • The biological effects of negatively-charged indoor air conditions 査読

    OTSUKI Takemi, TAKAHASHI Kazuaki, MASE Akinori, KAWADO Takashi, KOTANI Muneo, MAEDA Megumi, MURAKAMI Shuko, KUMAGAI Naoko, HAYASHI Hiroaki, CHEN Ying, NISHIMURA Yasumitsu, MIURA Yoshie, SHIRAHAMA Takashi, YOSHIMATSU Michiharu, MORIMOTO Kanehisa

    川崎医学会誌   35 ( 3 )   205 - 214   2009年10月

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    記述言語:英語   出版者・発行元:川崎医科大学  

    DOI: 10.11482/2009/35.205.2009_KMJ_Otsuki_etal.pdf

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  • Cytokine alteration and speculated immunological pathophysiology in silicosis and asbestos-related diseases. 査読 国際誌

    Shuko Murakami, Yasumitsu Nishimura, Megumi Maeda, Naoko Kumagai, Hiroaki Hayashi, Ying Chen, Masayasu Kusaka, Takumi Kishimoto, Takemi Otsuki

    Environmental health and preventive medicine   14 ( 4 )   216 - 22   2009年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    This review is partly composed of the presentation "Cytokine alteration and speculated immunological pathophysiology in silicosis and asbestos-related diseases" delivered during the symposium "Biological effects of fibrous and particulate substances and related areas" organized by the Study Group of Fibrous and Particulate Studies of the Japanese Society of Hygiene and held at the 78th Annual Meeting in Kumamoto, Japan. In this review, we briefly introduce the results of recent immunological analysis using the plasma of silica and asbestos-exposed patients diagnosed with silicosis, pleural plaque, or malignant mesothelioma. Thereafter, experimental background and speculation concerning the immunological pathophysiology of silica and asbestos-exposed patients are discussed.

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  • マイナス荷電粒子優位な室内空気環境の生体影響の観察 精神・神経・内分泌・免疫ネットワークへの影響について

    西村 泰光, 高橋 一聡, 間瀬 昭則, 河戸 隆, 小谷 宗男, 阿見 和久, 松島 弘樹, 白濱 毅, 吉松 道晴, 前田 恵, 村上 周子, 林 宏明, 熊谷 直子, 陳 瑩, 三浦 由恵, 森本 謙曩, 大槻 剛巳

    日本職業・環境アレルギー学会雑誌   16 ( 2 )   15 - 24   2009年5月

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    記述言語:日本語   出版者・発行元:日本職業・環境アレルギー学会  

    室内環境によって相互に関連する精神、神経、内分泌ならびに免疫の相互作用(PNEI(psycho-neuro-endocrino-immune)-ネットワーク)に影響するシックハウス症候群や化学物質過敏症が知られているが、室内環境の改善は健康増進という観点からも重要である。マイナス荷電粒子優位な室内環境を構築し、そのPNEIネットワークに及ぼす影響を検討したところ、時間単位の短期滞在試験では血清IL-2の上昇、週単位の中期滞在試験ではNK細胞の活性化が認められた。ストレス指標などには顕著な影響は認められなかったが、悪影響もなく今後月〜年単位の長期居住の影響あるいは細胞や遺伝子レベルへの影響の検討も興味深いと考えられた。また、森林浴などの健康効果の一部にもマイナス荷電粒子の影響が及んでいることも想定された。(著者抄録)

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  • Two weeks of permanence in negatively-charged air conditions causes alteration of natural killer cell function 査読

    K. Takahashi, Takemi Otsuki, Takemi Otsuki, A. Mase, T. Kawado, M. Kotani, Y. Nishimura, M. Maeda, S. Murakami, N. Kumagai, H. Hayashi, Y. Chen, T. Shirahama, Y. Miura, K. Morimoto

    International Journal of Immunopathology and Pharmacology   22   333 - 342   2009年1月

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    掲載種別:研究論文(学術雑誌)  

    The effects of negatively-charged air conditions were analyzed as one of the approaches to improve health and quality of life. We previously reported that the use of a charcoal coating and application of an electric voltage yielded predominantly negatively-charged particles in an experimental room, and that 2.5 hours of living in these conditions caused a slight activation of the immune system (slight elevation of serum interleukin (IL)-2), regulated blood flow, and stabilized the autonomic nervous system when compared with control conditions (no dominance of negatively-charged particles). In this study, we expanded the previous study and placed 15 subjects in negatively-charged air conditions for two weeks during the night and analyzed various biological parameters. Although individual biological reactions differed from subject to subject, natural killer (NK) cell activity increased significantly following living in negatively-charged air conditions. Taken together, the results of the previous investigation and those of this study show that repeated elevation of IL-2 (although it immediately returned to the baseline level) causes chronic and recurrent stimulation to NK cells and results in the steady activation of NK cells. Negatively-charged air particles may be a good tool to improve health and quality of life. Copyright © by Biolife, s.a.s.

    DOI: 10.1177/039463200902200210

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  • Effects of vitamin D receptor gene polymorphisms on low-resistance training using exercise machines: the 'Power Rehabilitation' program. 査読 国際誌

    Shin-Ichiro Murakami, Takemi Otsuki, Megumi Maeda, Yoshie Miura, Seiko Morii, Kenji Kiyokane, Shin-Ichi Hayakawa, Atsushi Maeda, Takayo Imakawa, Shunpei Harada, Torataro Handa, Yasumitsu Nishimura, Shuko Murakami, Naoko Kumagai, Hiroaki Hayashi, Ying Chen, Shin-Ichiro Suemori, Yumiko Fukushima, Seikoh Nishida, Keisuke Fukushima

    International journal of molecular medicine   23 ( 1 )   81 - 8   2009年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The enhancement and promotion of health is necessary to maintain the quality of life (QOL) of the aged population in developed nations such as Japan where the number of elderly has been increasing rapidly. For this purpose, low-resistance training using exercise machines ('Power Rehabilitation') has been established as a rehabilitation program. To investigate the individual factors which influence the effects of 'Power Rehabilitation', single nucleotide polymorphisms (SNPs) in the vitamin D receptor (VDR) gene and the ciliary neurotrophic factor (CNTF) gene were analyzed, and the relationship between SNP patterns and the effects of 'Power Rehabilitation' was evaluated. 'Power Rehabilitation' had an effect on the physiological functions involved in the activities of daily life (ADL) rather than muscle strength and size. In addition, certain SNP patterns showed better improvement of parameters associated with the effects of 'Power Rehabilitation' as analyzed by comparison between SNP patterns and factor analysis. Large scale analyses are required to ensure this tendency and to discover individual factors which may help to promote the health and QOL of the aged population.

    DOI: 10.3892/ijmm_00000104

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  • Decrease in phosphorylation of ERK following decreased expression of NK cell-activating receptors in human NK cell line exposed to asbestos 査読

    Yasumitsu Nishimura, M. Maeda, N. Kumagai, H. Hayashi, Y. Miura, T. Otsuki

    International Journal of Immunopathology and Pharmacology   22 ( 4 )   879 - 888   2009年

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    掲載種別:研究論文(学術雑誌)  

    YT-CB5, which had been continuously cultured with chrysotile B (CB) asbestos, showed impaired cytotoxicity with decreased expression of NKG2D and 2B4 NK cell-activating receptors. In the present study, the phosphorylation of extracellular signal-regulated kinase (ERK), which is known to induce degranulation downstream of many NK cell-activating receptors, was examined in YT-CB5 by flow cytometry and compared with the control line YT-Org. YT-CB5 exhibited impaired phosphorylation of ERK1/2 induced by the recognition of K562 cells, downstream of a process mediated by Src family kinase and phosphoinositide 3-kinase. YT-CBS also exhibited impaired phosphorylation of ERK1/2 following incubation with K562 cells in the presence of anti-2B4 antibodies, where co-stimulation by 2B4 augmented the phosphorylation of ERK1/2 in YT-CB5 to a similar degree as in YT-Org. The phosphorylation of ERK1/2 induced by an inhibitor against phosphatase (PP) 1 and PP2A was also lower in YT-CB5 compared with YT-Org. Moreover, bead-bound antibodies to NKG2D, which contribute to cytotoxicity against K562 cells, induced negligible phosphorylation of ERK1/2 in YT-CB5, although antibodies to 2B4. induced a comparatively greater level of phosphorylation. Additionally, peripheral blood (PB-) NK cells with low expression of NKG2D showed lower phosphorylation of ERK1/2 mediated by anti-NKG2D antibodies compared with PB-NK cells with high expression of NKG2D. These results indicate that signal transduction events leading to the phosphorylation of ERK is impaired in YT-CB5 due to decreased expression of NKG2D. Further studies are required to clarify whether this suppressive effect of asbestos exposure on NK cells might promote lung cancer and mesothelioma in people who have inhaled asbestos. Copyright © by Biolife, s.a.s.

    DOI: 10.1177/039463200902200403

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  • 珪肺症例末梢血CD4+25+分画における慢性活性化T細胞の混入とFas分子発現

    林 宏明, 前田 恵, 村上 周子, 西村 泰光, 大槻 剛巳, 藤本 亘

    山口医学   57 ( 4 )   111 - 112   2008年8月

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    記述言語:日本語   出版者・発行元:山口大学医学会  

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  • 繊維・粒子状物質の生体影響とその周辺 アスベストの免疫系への影響と腫瘍免疫減衰

    大槻 剛巳, 熊谷 直子, 村上 周子, 前田 恵, 林 宏明, 西村 泰光

    日本衛生学雑誌   63 ( 2 )   236 - 237   2008年3月

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    記述言語:日本語   出版者・発行元:(一社)日本衛生学会  

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  • Y2-4 アスベストの免疫影響の検討を踏まえたアスベスト曝露症例における免疫担当細胞の変化(胸膜疾患に対する診断の開発,要望演題2,第31回日本呼吸器内視鏡学会学術集会)

    大槻 剛巳, 前田 恵, 三浦 由恵, 村上 周子, 林 弘明, 熊谷 直子, 西村 泰光, 岸本 卓巳, 中野 孝司

    気管支学   30   S114   2008年

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    記述言語:日本語   出版者・発行元:特定非営利活動法人 日本呼吸器内視鏡学会  

    DOI: 10.18907/jjsre.30.Special_S114

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  • 珪肺症例におけるCD4+25+分画における慢性活性化T細胞、Fas分子発現の検討

    林 宏明, 前田 恵, 村上 周子, 西村 泰光, 草加 勝康, 大槻 剛巳

    日本臨床免疫学会総会抄録集   35   103 - 103   2007年

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    記述言語:日本語   出版者・発行元:日本臨床免疫学会  

    珪肺症の合併症として自己免疫疾患の頻度が高いことは成書にも記されている。CD4+25+FoxP3+制御性T細胞(Treg)が,自己寛容維持に重要な役割を有することが知られてきた。CD4+25+ Treg の特異的発現機能遺伝子であるFoxP3 の発現が,健常人では,CD4+25+分画で高発現,CD4+25-分画で極めて低発現なのに比し,珪肺症例ではCD4+25+分画でも,CD4+25-分画と同程度に低発現なことを以前に報告した。活性化T細胞では発現しているが,CD4+25+Treg細胞では発現していないPD-1遺伝子の発現を,珪肺症例と健常人の末梢血単核球をCD4+25+分画とCD4+25-分画において検討した。結果,健常人ではどちらの分画でも非常に低発現であったが,珪肺症例では,CD4+25-分画でも健常人より高度に,CD4+25+分画では更に高度な発現が認められた。また、CD4+FoxP3+分画のFasの発現は健常人においてもCD4+FoxP3-分画に比し高発現の度合いが高かった。これらの結果より,珪肺症例のCD4+25+分画には珪酸曝露に伴う慢性活性化T細胞が侵入し,および珪肺症例の制御性T細胞では,Fas発現が高く容易にアポトーシスに至る可能性が示唆された。その結果FoxP3+ Tregを置換,CD4+25+分画の機能の減衰をもたらして自己寛容の破綻を招いている可能性が高いと考えられた。

    DOI: 10.14906/jscisho.35.0.103.0

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  • 206 マンノース結合レクチンのスギ花粉症における役割(自然免疫, 第55回日本アレルギー学会秋季学術大会)

    岡野 光博, 前田 恵, 村井 綾, 藤原 田鶴子, 木村 吉伸, 西崎 和則

    アレルギー   54 ( 8 )   1058 - 1058   2005年

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    記述言語:日本語   出版者・発行元:一般社団法人 日本アレルギー学会  

    DOI: 10.15036/arerugi.54.1058_2

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  • 291 遊離型糖鎖によるCryj1特異的T細胞応答の制御

    岡野 光博, 木村 吉伸, 紀 光助, 木村 万里子, 菅田 裕士, 道上 佳洋, 前田 恵, 松田 史子, 西崎 和則

    アレルギー   51 ( 9 )   979 - 979   2002年

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    記述言語:日本語   出版者・発行元:一般社団法人 日本アレルギー学会  

    DOI: 10.15036/arerugi.51.979_3

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  • 293 遊離型糖鎖を用いたCryj1構成糖鎖のIgEエピトープ活性に関する検討

    菅田 裕士, 岡野 光博, 木村 吉伸, 紀 光助, 木村 万里子, 道上 佳洋, 酒本 真次, 前田 恵, 西崎 和則

    アレルギー   51 ( 9 )   980 - 980   2002年

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    記述言語:日本語   出版者・発行元:一般社団法人 日本アレルギー学会  

    DOI: 10.15036/arerugi.51.980_1

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書籍等出版物

  • Sugar Chain Analysis by Enzymatic Digenstion and 2D Mapping by HPLC.. Experimental Glycoscience, Glycochemistry

    Springer 

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  • Sugar Chain Analysis by Enzymatic Digenstion and 2D Mapping by HPLC.「Experimental Glycoscience, Glycochemistry」

    Springer 

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MISC

  • Ginkgo biloba α-fucosidase with activity towards plant complex type N-glycans containing the Lewis a epitope: Purification and characterization

    Satsuki Itano, Megumi Maeda, Md. Ziaur Rahman, Yoshinobu Kimura

    岡山大学農学部学術報告   106   5 - 12   2017年

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    記述言語:英語   出版者・発行元:岡山大学農学部  

    We have identified, and purified to homogeneity, a high molecular weight Ginkgo biloba α-fucosidase (α-fucosidase Gb, 120 kDa estimated by SDS‒PAGE) with activity against α-fucosylated oligosaccharides. When a Lewis a epitope-containing N-glycan was used as a substrate, α-fucosidase Gb showed optimum activity at approximately pH 5.5, suggesting that it functions in acidic environments such as the vacuole. It remains uncertain, however, whether this Ginkgo α-fucosidase belongs to the GH29 family, since its N-terminal sequence could not be determined, probably due to a chemical modification. α-Fucosidase Gb showed substantial activity towards the α1,3-fucosyl linkage in Lacto-N-fucopentaose III and an α1,4-fucosyl linkage in the Lewis a epitope found in plant complex type N-glycans, indicating an involvement in the degradation process of α-fucosylated oligosaccharides or N-glycoproteins. 銀杏種子から高分子量 (SDS-PAGE で120 kDa) を有し,α-フコース含有オリゴ糖に活性を示すα-フコシダーゼ(α-fucosidase Gb)を均一に精製した.ルイス a エピトープ含有 N-グリカンを基質とした場合,α-fucosidase Gb の至適 pH は 5.5 付近であることから,本酵素は液胞のような酸性環境で機能していることが示唆された.N‒末端アミノ酸配列が化学修飾のため同定できなかったため,本酵素が GH29 ファミリーに属するかどうかは不明である.α-Fucosidase Gb は,Lacto-N-fucopentaose IIIの α1,3-フコース残基やルイス a エピトープ含有の植物複合型N-グリカンのα1,4-フコース残基を加水分解することから,α-フコース含有オリゴ糖やN 型糖タンパク質の分解プロセスに関与することが示唆された.

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  • Structural features of free N-glycans occurring in plants and functional features of de-N-glycosylation enzymes, ENGase, and PNGase: the presence of unusual plant complex type N-glycans 国際誌

    Megumi Maeda, Yoshinobu Kimura

    FRONTIERS IN PLANT SCIENCE   5   429 - 429   2014年9月

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    記述言語:英語   出版者・発行元:FRONTIERS RESEARCH FOUNDATION  

    Free N-glycans (FNGs) are present at micromolar concentrations in plant cells during their differentiation, growth, and maturation stages. It has been postulated that these FNGs are signaling molecules involved in plant development or fruit ripening. However, the hypothetical biochemical and molecular function of FNGs has not been yet established.The structure of FNGs found ubiquitously in plant tissues such as hypocotyls, leaves, roots, developing seeds, or fruits can be classified into two types: high-mannose type and plant complex type; the former, in most cases, has only one GIcNAc residue at the reducing end (GNI type), while the latter has the chitobiosyl unit at the reducing end (GN2 type). These findings suggest that endo-13-N-acetylglucosaminidase (ENGase) must be involved in the production of GN1 type FNGs, whereas only peptide:N-glycanase (PNGase) is involved in the production of GN2 type FNGs. It has been hypothesized that cytosolic PNGase (cPNGase) and ENGase in animal cells are involved in the production of high-mannose type FNGs in order to release N-glycans from the misfolded glycoproteins in the protein quality control systems. In the case of plants, it is well known that another type of PNGase, the acidic PNGase (aPNGase) is involved in the production of plant complex type FNGs in an acidic organelle, suggesting the de-N-glycosylation mechanism in plants is different from that in animal cells. To better understand the role of these FNGs in plants, the genes encoding these N-glycan releasing enzymes (ENGase and PNGase) were first identified, and then structure of FNGs in ENGase knocked-out plants were analyzed.These transgenic plants provide new insight into the plant-specific de-N-glycosylation mechanism and putative physiological functions of FNGs. In this review, we focus on the structural features of plant FNGs, as well as functional features of cPNGase/ENGase and plant specific PNGase, and putative functions of FNGs are also discussed.

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  • Changes in Glycinin‒Digesting Protease Activity During Soybean Germination.

    Md. Akhtaruzzaman, Maeda Megumi, Kitagawa Keiko, Takagi Shigeaki, Kimura Yoshinobu

    岡山大学農学部学術報告   103   1 - 4   2014年2月

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    記述言語:英語   出版者・発行元:岡山大学農学部  

    Changes in glycinin-digesting protease activity during soybean germination have been investigated.The glycinin-digesting protease activities of imbibed or germinated soybean seed were assayed byRP‒HPLC using a tryptic peptide from CM‒glycinin or by SDS‒PAGE using CM‒glycinin as the endogenoussubstrate. Proteolytic activities of the germinated soybean seeds were found through the wholeperiod of germination, the activities were maintained significantly unchanged during germination for 4days, and then those specific activities declined slowly. AE‒HPLC analysis of the glycinin-digestingprotease in the imbibed or germinated soybean seeds showed unchanged peaks corresponding to glycinin-digesting activity, suggesting that the glycinin-digesting protease was not induced during germinationbut had already been synthesized during seed maturation. 大豆発芽期におけるグリシニン分解酵素 (98 kDa SBP) の活性変動を解析した.大豆種子を4時間水で膨潤後, 25℃ 暗黒下で発芽させた.経時的にサンプリングを行い,2M NaCl を含むトリス緩衝液 (㏗ 7.0) により粗酵素を抽出後,グリシニン由来のトリプシン分解ペプチドを基質としてグリシニン分解酵素の活性変動を逆相 HPLC により追跡した.その結果,種子膨潤後4日間比活性はほぼ一定の値を保ち,以後徐々に低下することが分かった.次いで,粗酵素溶液からイオン交換 HPLC により98 kDa SBP を部分精製するとともに,発芽期における 98 kDa SBP の消長を解析したところ,98 kDa SBP は乾燥種子及び各発芽段階の種子中全てに認められ,かつグリシン分解活性もグリシニン由来のトリプシン分解ペプチド基質に対する活性と同様に認められた.以上の結果から,98 kDa SBP は種子発芽に伴い誘導されるプロテアーゼではなく,種子貯蔵型のプロテアーゼであることが明らかになった.

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  • Suppression of cytosolic and acidic peptide: N-glycanase (PNGase) in Arabidopsis thaliana

    Tsuyoshi Akiyama, Megumi Maeda, Yoshinobu Kimura

    GLYCOBIOLOGY   23 ( 11 )   1378 - 1378   2013年11月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:OXFORD UNIV PRESS INC  

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  • Effect of EUL-related nucleocytoplasmic lectin from Arabidopsis thaliana, ArathEULS3, on the root growth under salt stress

    Megumi Maeda, Jonas Van Hove, Elke Fouquaert, Yoshinobu Kimura, Els Van Damme

    GLYCOBIOLOGY   23 ( 11 )   1364 - 1364   2013年11月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:OXFORD UNIV PRESS INC  

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  • 植物糖タンパク質の代謝に関わる糖鎖関連酵素の機能特性と遊離型糖鎖の存在意義 -植物endo-β-N-acetylglucosaminidase (ENGase) とpeptide:N-glycanase (PNGase)-

    前田恵, 木村吉伸

    応用糖質科学   3 ( 1 )   77 - 86   2013年

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    記述言語:日本語   出版者・発行元:日本応用糖質科学会  

    真核生物が産生する60%程度のタンパク質がグリコシル化を受けた糖タンパク質であることはすでに周知の事実である。しかしながら,分泌型タンパク質に限ってみるならば,それらのほとんどはこの糖タンパク質の範疇に入る。哺乳動物と高等植物では,それら糖タンパク質に結合するオリゴ糖鎖の構造特性にさまざまな違いがみられるが,それらの差異はゴルジ装置中で機能する糖鎖代謝酵素群(糖転移酵素と加水分解酵素)の基質特異性の違い,あるいはゲノム中にコードされる糖鎖代謝酵素群の遺伝子そのものの違いが反映されたものである。糖タンパク質糖鎖のうちアスパラギン結合型糖鎖(N-グリカン)に注目すると,それらの生合成およびプロセシング機構には真核生物に共通するコンテクストが存在し,構造多様性の中にも共通のトリマンノシルコア構造(Man3GlcNAc2-Asn,Man,マンノース;GlcNAc,N-アセチルグルコサミン,Asn,アスパラギン)がみられるなど,高等植物と哺乳動物とで構造特性の比較が可能である。哺乳動物と高等植物が産生するN-グリカンの代表的な構造を図1に示している。日南乳動物の複合型構造には極めて高い多様性があるので,わずかの例を挙げるにとどめるが,基本的には,(1)非還元末端にシアル酸残基が存在する場合が多い,(2)ガラクトース(Gal)残基は戸1-4結合でGlcNAc残基に結合する,(3)還元末端GlcNAc残基へのフコース(Fuc)残基の結合がα1-6結合である,(4)バイセクトGlcNAc残基がs1-4Man残基に結合する場合がある,(5)2本鎖構造以外に3~5本鎖構造が存在する等々が哺乳動物N-グリカンの構造的特徴である。一方,植物N-グリカンには(1)コア構造中のβ-Man残基ヘキシロース(Xyl)残基がβ1-2結合し,還元末端GlcNAc残基へFuc残基がα1-3結合する,(2)非還元末端側にルイスa(Lewis a,Le a)抗原(Galβ1-3(Fucα1-4)GlcNAc)が存在する場合があり,Gal残基はβ1-3結合でGlcNAc残基に結合する,(3)3~5本鎖構造やシアル酸を有する構造は見出されていない等の特徴がある。

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  • Large-Scale Preparation of Asn-Glycopeptide Carrying Structurally Homologous Antigenic N-Glycan 国際誌

    Megumi MAEDA, Naoto TAKEDA, Aya MANO, Maomi YAMANISHI, Mariko KIMURA, Yoshinobu KIMURA

    Bioscience, Biotechnology, and Biochemistry   77 ( 6 )   1269 - 1274   2013年

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    In our previous report (M. Okano, et al., Clin. Exp. Allergy, 34, 770-778 (2004)), we found that free plant complex type N-glycans suppressed the production of IL4 from Th2 cells of Japanese cedar pollinosis patients, suggesting that plant complex type N-glycan can be used as a leading compound for developing immuno-pharmaceuticals. Although immunoreactive plant complex type N-glycans occur ubiquitously on glycoproteins expressed in plants, an appropriate procedure has not been established to prepare non-labeled immunoreactive glycans or glycopeptides bearing structurally homologous immunoreactive glycans in large amounts. In this study, therefore, we developed a new preparative procedure for the large-scale preparation of Asn-glycopeptide bearing plant complex type N-glycan using a combination of gel-filtration and the hydrophilic partitioning method. By this new method, about 103 mg of Asn-glycopeptide bearing the antigenic N-glycans was obtained from 1.9 kg of shelled Ginkgo biloba seeds.

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  • [Archives of "comprehensive approach on asbestos-related diseases" supported by the "special coordination funds for promoting science and technology (H18-1-3-3-1)"-- overview of group research project, care and specimen registration, cellular characteristics of mesothelioma and immunological effects of asbestos].

    Takemi Otsuki, Takashi Nakano, Seiki Hasegawa, Morihito Okada, Tohru Tsujimura, Yoshitaka Sekido, Shinya Toyokuni, Hiroshi Nishimoto, Kazuya Fukuoka, Fumihiro Tanaka, Naoko Kumagai, Megumi Maeda, Yasumitsu Nishimura

    Nihon eiseigaku zasshi. Japanese journal of hygiene   66 ( 3 )   543 - 52   2011年5月

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    記述言語:日本語  

    The research project entitled "Comprehensive approach on asbestos-related diseases" supported by the "Special Coordination Funds for Promoting Science and Technology (H18-1-3-3-1)" began in 2006 and was completed at the end of the Japanese fiscal year of 2010. This project included four parts; (1) malignant mesothelioma (MM) cases and specimen registration, (2) development of procedures for the early diagnosis of MM, (3) commencement of clinical investigations including multimodal approaches, and (4) basic research comprising three components; (i) cellular and molecular characterization of mesothelioma cells, (ii) immunological effects of asbestos, and (iii) elucidation of asbestos-induced carcinogenesis using animal models. In this special issue of the Japanese Journal of Hygiene, we briefly introduce the achievements of our project. The second and third parts and the third component of the fourth part are described in other manuscripts written by Professors Fukuoka, Hasegawa, and Toyokuni. In this manuscript, we introduce a brief summary of the first part "MM cases and specimen registration", the first component of the fourth part "Cellular and molecular characterization of mesothelioma cells" and the second component of the fourth part "Immunological effects of asbestos". In addition, a previous special issue presented by the Study Group of Fibrous and Particulate Substances (SGFPS) (chaired by Professor Otsuki, Kawasaki Medical School, Japan) for the Japanese Society of Hygiene and published in Environmental Health and Preventive Medicine Volume 13, 2008, included reviews of the aforementioned first component of the fourth part of the project. Taken together, our project led medical investigations regarding asbestos and MM progress and contributed towards the care and examination of patients with asbestos-related diseases during these five years. Further investigations are required to facilitate the development of preventive measures and the cure of asbestos-related diseases, particularly in Japan, where asbestos-related diseases are predicted to increase in the next 10 to 20 years.

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  • 科学技術振興調整費「アスベスト関連疾患への総括的取り組み」班研究 成果報告 班研究の概要と登録事業、中皮腫細胞特性ならびにアスベストの免疫影響について

    大槻 剛巳, 中野 孝司, 長谷川 誠紀, 岡田 守人, 辻村 亨, 関戸 好孝, 豊國 伸哉, 西本 寛, 福岡 和也, 田中 文啓, 熊谷 直子, 前田 恵, 松崎 秀紀, 李 順姫, 西村 泰光, 科学技術振興調, 年度「アスベスト関連疾患への総括的取り組み, 研究班

    日本衛生学雑誌   66 ( 3 )   543 - 552   2011年5月

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    記述言語:日本語   出版者・発行元:(一社)日本衛生学会  

    The research project entitled "Comprehensive approach on asbestos-related diseases" supported by the "Special Coordination Funds for Promoting Science and Technology (H18-1-3-3-1)" began in 2006 and was completed at the end of the Japanese fiscal year of 2010. This project included four parts; (1) malignant mesothelioma (MM) cases and specimen registration, (2) development of procedures for the early diagnosis of MM, (3) commencement of clinical investigations including multimodal approaches, and (4) basic research comprising three components; (i) cellular and molecular characterization of mesothelioma cells, (ii) immunological effects of asbestos, and (iii) elucidation of asbestos-induced carcinogenesis using animal models. In this special issue of the Japanese Journal of Hygiene, we briefly introduce the achievements of our project. The second and third parts and the third component of the fourth part are described in other manuscripts written by Professors Fukuoka, Hasegawa, and Toyokuni. In this manuscript, we introduce a brief summary of the first part "MM cases and specimen registration", the first component of the fourth part "Cellular and molecular characterization of mesothelioma cells" and the second component of the fourth part "Immunological effects of asbestos". In addition, a previous special issue presented by the Study Group of Fibrous and Particulate Substances (SGFPS) (chaired by Professor Otsuki, Kawasaki Medical School, Japan) for the Japanese Society of Hygiene and published in Environmental Health and Preventive Medicine Volume 13, 2008, included reviews of the aforementioned first component of the fourth part of the project. Taken together, our project led medical investigations regarding asbestos and MM progress and contributed towards the care and examination of patients with asbestos-related diseases during these five years. Further investigations are required to facilitate the development of preventive measures and the cure of asbestos-related diseases, particularly in Japan, where asbestos-related diseases are predicted to increase in the next 10 to 20 years.<br>

    DOI: 10.1265/jjh.66.543

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  • 科学技術振興調整費「アスベスト関連疾患への総括的取り組み」班 登録事業の現状と、基礎研究よりアスベストの免疫系への影響について

    大槻 剛巳, 西本 寛, 前田 恵, 熊谷 直子, 林 宏明, 西村 泰光, 長谷川 誠紀, 田中 文啓, 岡田 守人, 福岡 和也, 辻村 亨, 関戸 好孝, 豊國 伸哉, 岸本 卓巳, 中野 孝司, 科学技術振興調整費, アスベスト関連疾患への総括的取り組み

    日本職業・災害医学会会誌   57 ( 臨増 )   別149 - 別149   2009年10月

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  • Molecular Cloning and Gene Expression Analysis of Tomato Endo-beta-N-acetylglucosaminidase, an Endoglycosidase Involved in the Production of High-Mannose Type Free N-Glycans during Tomato Fruit Ripening 国際誌

    Kosuke Nakamura, Masami Inoue, Megumi Maeda, Ryohei Nakano, Katsutoshi Hosoi, Kazuhito Fujiyama, Yoshinobu Kimura

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   73 ( 2 )   461 - 464   2009年2月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    In this study, we identified a gene encoding tomato ENGase (Endo-LE) using the gene information of rice ENGase, and expressed the Endo-LE protein in Escherichia coli. The substrate specificity of the recombinant Endo-LE was the same as that of the native enzyme, showing strong activity towards the high-mannose type N-glycans with the Man alpha 1-2Man alpha 1-3Man beta 1-4GlcNAc beta 1-4GlcNAc unit. Real-time PCR analysis revealed that the gene expression of Endo-LE did not vary significantly with the tomato ripening process, indicating that Endo-LE activity is ubiquitously expressed.

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  • Molecular Cloning and Gene Expression Analysis of Tomato Endo-beta-N-acetylglucosaminidase, an Endoglycosidase Involved in the Production of High-Mannose Type Free N-Glycans during Tomato Fruit Ripening

    Kosuke Nakamura, Masami Inoue, Megumi Maeda, Ryohei Nakano, Katsutoshi Hosoi, Kazuhito Fujiyama, Yoshinobu Kimura

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   73 ( 2 )   461 - 464   2009年2月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    In this study, we identified a gene encoding tomato ENGase (Endo-LE) using the gene information of rice ENGase, and expressed the Endo-LE protein in Escherichia coli. The substrate specificity of the recombinant Endo-LE was the same as that of the native enzyme, showing strong activity towards the high-mannose type N-glycans with the Man alpha 1-2Man alpha 1-3Man beta 1-4GlcNAc beta 1-4GlcNAc unit. Real-time PCR analysis revealed that the gene expression of Endo-LE did not vary significantly with the tomato ripening process, indicating that Endo-LE activity is ubiquitously expressed.

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  • Soluble interleukin-2 receptor as an indicator of immunological disturbance found in silicosis patients

    H. Hayashi, M. Maeda, S. Murakami, N. Kumagai, Y. Chen, T. Hatayama, M. Katoh, N. Miyahara, S. Yamamoto, Y. Yoshida, Y. Nishimura, M. Kusaka, W. Fujimoto, Takemi Otsuki

    International Journal of Immunopathology and Pharmacology   22 ( 1 )   53 - 62   2009年

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    記述言語:英語   出版者・発行元:Biomedical Research Press s.a.s.  

    Silicosis patients (SILs) possess not only respiratory disorders but also alterations in autoimmunity. To determine an early indicator of immunological disturbance in SILs, the role of serum-soluble interleukin (IL)-2 receptor (sIL-2R) was analyzed. Of ten SlLs, immunological clinical parameters such as immunoglobulin (Ig) G, complements, the titer of autoantibodies including anti-nuclear antibodies (ANA), anti-Scl-70 antibody (Ab) and anti-centromere (CM) Ab, and experimental indicators such as serum-soluble Fas, serum IL-2, CD25+ cells in CD4+ or CD8+ fractions, and sIL-2R were divided from respiratory parameters such as % vital capacity (%VC), percentage of forced expiratory volume in 1 second (FEV1.0%) and v25/Ht (liter/second/m(body height) by a correlation assay. Additionally, a stepwise regression test showed that sIL-2R was correlated with Ig G, ANA and anti-CM Ab. Furthermore, factor analysis revealed that sIL-2R contributed to the subpopulation of SILs with poorer immunological status in the absence of alterations in respiratory status. By defining healthy donors as 1, SILs as 2 and patients with systemic sclerosis as 3 for immunopathological progression status as metric variables, sIL2R and ANA showed a strong positive correlation. This suggests that sIL-2R is a good clinical indicator of immunological disturbance found in SILs without clinical manifestations of any disturbance in autoimmunity. Further analysis using a large-scale number of patients should be performed to confirm these findings. Copyright © by BIOLIFE.

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  • Soluble interleukin-2 receptor as an indicator of immunological disturbance found in silicosis patients

    H. Hayashi, M. Maeda, S. Murakami, N. Kumagai, Y. Chen, T. Hatayama, M. Katoh, N. Miyahara, S. Yamamoto, Y. Yoshida, Y. Nishimura, M. Kusaka, W. Fujimoto, Takemi Otsuki

    International Journal of Immunopathology and Pharmacology   22 ( 1 )   53 - 62   2009年

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    記述言語:英語   出版者・発行元:Biomedical Research Press s.a.s.  

    Silicosis patients (SILs) possess not only respiratory disorders but also alterations in autoimmunity. To determine an early indicator of immunological disturbance in SILs, the role of serum-soluble interleukin (IL)-2 receptor (sIL-2R) was analyzed. Of ten SlLs, immunological clinical parameters such as immunoglobulin (Ig) G, complements, the titer of autoantibodies including anti-nuclear antibodies (ANA), anti-Scl-70 antibody (Ab) and anti-centromere (CM) Ab, and experimental indicators such as serum-soluble Fas, serum IL-2, CD25+ cells in CD4+ or CD8+ fractions, and sIL-2R were divided from respiratory parameters such as % vital capacity (%VC), percentage of forced expiratory volume in 1 second (FEV1.0%) and v25/Ht (liter/second/m(body height) by a correlation assay. Additionally, a stepwise regression test showed that sIL-2R was correlated with Ig G, ANA and anti-CM Ab. Furthermore, factor analysis revealed that sIL-2R contributed to the subpopulation of SILs with poorer immunological status in the absence of alterations in respiratory status. By defining healthy donors as 1, SILs as 2 and patients with systemic sclerosis as 3 for immunopathological progression status as metric variables, sIL2R and ANA showed a strong positive correlation. This suggests that sIL-2R is a good clinical indicator of immunological disturbance found in SILs without clinical manifestations of any disturbance in autoimmunity. Further analysis using a large-scale number of patients should be performed to confirm these findings. Copyright © by BIOLIFE.

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  • Impairment in cytotoxicity and expression of NK cell-activating receptors on human NK cells following exposure to asbestos fibers

    Yasumitsu Nishimura, Y. Miura, M. Maeda, N. Kumagai, S. Murakami, H. Hayashi, K. Fukuoka, T. Nakano, T. Otsuki

    International Journal of Immunopathology and Pharmacology   22 ( 3 )   579 - 590   2009年

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    Asbestos is well-known for its tumorigenic activity, but its effect on anti-tumor immunity remains unclear. Therefore, we prepared a sub-line of YT-A1 human NK cells exposed to chrysotile B (CB) asbestos (YT-CB5) as an in vitro model to analyze the effect of asbestos exposure on NK cells, and examined cytotoxicity and expressions of its related molecules. The cytotoxicity of YT-CB5 against K562 cells decreased compared with the original line of YT-A1 (YT-Org). YT-CB5 exhibited significant decreases in expressions of cell surface NKG2D, 2B4 and intracellular granzyme A. YT-CB5 also exhibited a decrease in the 2B4-dependent cytotoxicity. In addition, the degranulations stimulated via cell surface NKG2D and 2B4 also decreased in YT-CB5. Therefore, peripheral blood NK cells in patients with malignant mesothelioma (MM) were examined and compared with healthy volunteers. NK cells in patients with MM also showed decreases in cytotoxicity against K562. Although the expressions of NKG2D and 2B4 did not decrease in NK cells of MM patients, the expression of cell surface NKp46 decreased. To confirm the effect of asbestos exposure on peripheral blood NK cells, PBMCs were cultured under exposure to CB. NK cells in PBMCs exposed to CB in vitro showed a significant decrease in the expression of NKp46, whereas NK cells in PBMCs exposed to glass wool did not show such a decrease. These results indicate that exposure to asbestos has the potential to impair the cytotoxicity of NK cells and alter the expression of NK cell-activating receptors including NKG2D, 2B4 and NKp46 and intracellular perforin/granzymes. Copyright © by BIOLIFE, s.a.s.

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  • P-39 悪性中皮腫患者NK細胞におけるNKp46発現量低下と細胞傷害性低下の相関性(基礎研究,第49回日本肺癌学会総会号)

    西村 泰光, 前田 恵, 村上 周子, 熊谷 直子, 林 宏明, 三浦 由恵, 岸本 卓巳, 大槻 剛巳

    肺癌   48 ( 5 )   499 - 499   2008年10月

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    記述言語:日本語   出版者・発行元:日本肺癌学会  

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  • P-665 アスベスト曝露症例における実験的な影響の観察を踏まえた免疫担当細胞の変化(悪性中皮腫3,第49回日本肺癌学会総会号)

    大槻 剛巳, 前田 恵, 村上 周子, 熊谷 直子, 林 宏明, 栗林 康造, 福岡 和也, 岸本 卓巳, 中野 孝司, 西村 泰光

    肺癌   48 ( 5 )   656 - 656   2008年10月

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  • Negatively-charged air conditions and responses of the human psycho-neuro-endocrino-immune network. 国際誌

    Kazuaki Takahashi, Takemi Otsuki, Akinori Mase, Takashi Kawado, Muneo Kotani, Kazuhisa Ami, Hiroki Matsushima, Yasumitsu Nishimura, Yoshie Miura, Shuko Murakami, Megumi Maeda, Hiroaki Hayashi, Naoko Kumagai, Takashi Shirahama, Michiharu Yoshimatsu, Kanehisa Morimoto

    Environment international   34 ( 6 )   765 - 72   2008年8月

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    BACKGROUND: Against increasing environmental adverse effects on human health such as those associated with water and ground pollution, as well as out- and indoor air conditions, trials were conducted to support and promote human health by improving the indoor air atmosphere. This study was performed to estimate the effect of negatively-charged air conditions on human biological markers related to the psycho-neuro-endocrino-immune (PNEI) network. OBJECTIVES: After construction of negatively-charged experimental rooms (NCRs), healthy volunteers were admitted to these rooms and control rooms (CTRs) and various biological responses were analyzed. METHODS: NCRs were constructed using a fine charcoal coating and applying an electric voltage (72 V) between the backside of walls and the ground. Various biological markers were monitored that related to general conditions, autonomic nervous systems, stress markers, immunological parameters and blood flow. RESULTS: Regarding the indoor environment, only negatively-charged air resulted in the difference between the CTR and NCR groups. The well-controlled experimental model-room to examine the biological effects of negatively-charged air was therefore established. Among the various parameters, IL-2, IL-4, the mean RR interval of the heart rate, and blood viscosity differed significantly between the CTR and NCR groups. In addition, the following formula was used to detect NCR-biological responses: Biological Response Value (BRV)=0.498+0.0005 [salivary cortisol]+0.072 [IL-2]+0.003 [HRM-SD]-0.013 [blood viscosity]-0.009 [blood sugar]+0.017 [pulse rate]. CONCLUSIONS: Negatively-charged air conditions activated the immune system slightly, smoothened blood flow and stabilized the autonomic nervous system. Although this is the first report to analyze negatively-charged air conditions on human biological responses, the long-term effects should be analyzed for the general use of these artificial atmospheres.

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  • Negatively-charged air conditions and responses of the human psycho-neuro-endocrino-immune network. 国際誌

    Kazuaki Takahashi, Takemi Otsuki, Akinori Mase, Takashi Kawado, Muneo Kotani, Kazuhisa Ami, Hiroki Matsushima, Yasumitsu Nishimura, Yoshie Miura, Shuko Murakami, Megumi Maeda, Hiroaki Hayashi, Naoko Kumagai, Takashi Shirahama, Michiharu Yoshimatsu, Kanehisa Morimoto

    Environment international   34 ( 6 )   765 - 72   2008年8月

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    BACKGROUND: Against increasing environmental adverse effects on human health such as those associated with water and ground pollution, as well as out- and indoor air conditions, trials were conducted to support and promote human health by improving the indoor air atmosphere. This study was performed to estimate the effect of negatively-charged air conditions on human biological markers related to the psycho-neuro-endocrino-immune (PNEI) network. OBJECTIVES: After construction of negatively-charged experimental rooms (NCRs), healthy volunteers were admitted to these rooms and control rooms (CTRs) and various biological responses were analyzed. METHODS: NCRs were constructed using a fine charcoal coating and applying an electric voltage (72 V) between the backside of walls and the ground. Various biological markers were monitored that related to general conditions, autonomic nervous systems, stress markers, immunological parameters and blood flow. RESULTS: Regarding the indoor environment, only negatively-charged air resulted in the difference between the CTR and NCR groups. The well-controlled experimental model-room to examine the biological effects of negatively-charged air was therefore established. Among the various parameters, IL-2, IL-4, the mean RR interval of the heart rate, and blood viscosity differed significantly between the CTR and NCR groups. In addition, the following formula was used to detect NCR-biological responses: Biological Response Value (BRV)=0.498+0.0005 [salivary cortisol]+0.072 [IL-2]+0.003 [HRM-SD]-0.013 [blood viscosity]-0.009 [blood sugar]+0.017 [pulse rate]. CONCLUSIONS: Negatively-charged air conditions activated the immune system slightly, smoothened blood flow and stabilized the autonomic nervous system. Although this is the first report to analyze negatively-charged air conditions on human biological responses, the long-term effects should be analyzed for the general use of these artificial atmospheres.

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  • 【知っておきたい悪性中皮腫】 アスベスト関連疾患の発症機構 悪性中皮腫を中心として

    西村 泰光, 前田 恵, 村上 周子, 熊谷 直子, 林 宏明, 大槻 剛巳

    Surgery Frontier   15 ( 2 )   122 - 127   2008年6月

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    記述言語:日本語   出版者・発行元:(株)メディカルレビュー社  

    アスベスト関連疾患への発症機構としては、線維化と癌化の二方向を考慮しなければならない。線維化の代表疾患が石綿肺であり、癌化ではアスベスト関連肺癌や悪性中皮腫が該当する。線維化については、アスベスト繊維の長さと処理役としての肺胞マクロファージ、産生するサイトカインとの相互作用が重要である。癌化について悪性中皮腫をモデルに考えると、腫瘍抑制遺伝子として細胞周期にかかわるp16INK4a,p14ARF、細胞の接着や増殖にかかわる神経線維腫症2型遺伝子(NF-2)の欠失あるいは発現低下が知られている。また増殖因子としては、PDGF,HGF,IGFなどが関与するとともに、血管新生ではVEGFが重要性である。加えて、BCl-XLやテロメラーゼの変化による抗アポトーシスや不死化能の獲得も癌化には関与している。なお、アスベストによる腫瘍免疫の減弱も癌化に影響している可能性がある。(著者抄録)

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  • CRTH2 plays an essential role in the pathophysiology of Cry j 1-induced pollinosis in mice 国際誌

    Rie Nomiya, Mitsuhiro Okano, Tazuko Fujiwara, Megumi Maeda, Yoshinobu Kimura, Kosuke Kino, Minehiko Yokoyama, Hiroyuki Hirai, Kinya Nagata, Toshifumi Hara, Kazunori Nishizaki, Masataka Nakamura

    JOURNAL OF IMMUNOLOGY   180 ( 8 )   5680 - 5688   2008年4月

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    記述言語:英語   出版者・発行元:AMER ASSOC IMMUNOLOGISTS  

    PGD(2) is the major prostanoid produced during the acute phase of allergic reactions. Two PGD(2) receptors have been isolated, DP and CRTH2 (chemoattractant receptor-homologous molecule expressed on Th2 cells), but whether they participate in the pathophysiology of allergic diseases remains unclear. We investigated the role of CRTH2 in the initiation of allergic rhinitis in mice. First, we developed a novel murine model of pollinosis, a type of seasonal allergic rhinitis. Additionally, pathophysiological differences in the pollinosis were compared between wild-type and CRTH2 gene-deficient mice. An effect of treatment with ramatroban, a CRTH2/T-prostanoid receptor dual antagonist, was also determined. Repeated intranasal sensitization with Cry j 1, the major allergen of Cryptomeria japonica pollen, in the absence of adjuvants significantly exacerbated nasal hyperresponsive symptoms, Cry j 1-specific IgE and 1gG1 production, nasal eosinophilia, and Cry j 1-induced in vitro production of IL-4 and IL-5 by submandibular lymph node cells. Additionally, CRTH2 mRNA in nasal mucosa was significantly elevated in Cry j 1-sensitized mice. Following repeated intranasal sensitization with Cry j 1, CRTH2 gene-deficient mice had significantly weaker Cry j 1-specific IgE/IgG1 production, nasal eosinophilia, and IL-4 production by submandibular lymph node cells than did wild-type mice. Similar results were found in mice treated with ramatroban. These results suggest that the PGD(2)- CRTH2 interaction is elevated following sensitization and plays a proinflammatory role in the pathophysiology of allergic rhinitis, especially pollinosis in mice.

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  • CRTH2 plays an essential role in the pathophysiology of Cry j 1-induced pollinosis in mice

    Rie Nomiya, Mitsuhiro Okano, Tazuko Fujiwara, Megumi Maeda, Yoshinobu Kimura, Kosuke Kino, Minehiko Yokoyama, Hiroyuki Hirai, Kinya Nagata, Toshifumi Hara, Kazunori Nishizaki, Masataka Nakamura

    JOURNAL OF IMMUNOLOGY   180 ( 8 )   5680 - 5688   2008年4月

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    記述言語:英語   出版者・発行元:AMER ASSOC IMMUNOLOGISTS  

    PGD(2) is the major prostanoid produced during the acute phase of allergic reactions. Two PGD(2) receptors have been isolated, DP and CRTH2 (chemoattractant receptor-homologous molecule expressed on Th2 cells), but whether they participate in the pathophysiology of allergic diseases remains unclear. We investigated the role of CRTH2 in the initiation of allergic rhinitis in mice. First, we developed a novel murine model of pollinosis, a type of seasonal allergic rhinitis. Additionally, pathophysiological differences in the pollinosis were compared between wild-type and CRTH2 gene-deficient mice. An effect of treatment with ramatroban, a CRTH2/T-prostanoid receptor dual antagonist, was also determined. Repeated intranasal sensitization with Cry j 1, the major allergen of Cryptomeria japonica pollen, in the absence of adjuvants significantly exacerbated nasal hyperresponsive symptoms, Cry j 1-specific IgE and 1gG1 production, nasal eosinophilia, and Cry j 1-induced in vitro production of IL-4 and IL-5 by submandibular lymph node cells. Additionally, CRTH2 mRNA in nasal mucosa was significantly elevated in Cry j 1-sensitized mice. Following repeated intranasal sensitization with Cry j 1, CRTH2 gene-deficient mice had significantly weaker Cry j 1-specific IgE/IgG1 production, nasal eosinophilia, and IL-4 production by submandibular lymph node cells than did wild-type mice. Similar results were found in mice treated with ramatroban. These results suggest that the PGD(2)- CRTH2 interaction is elevated following sensitization and plays a proinflammatory role in the pathophysiology of allergic rhinitis, especially pollinosis in mice.

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  • Reduced function of the peripheral CD4+25+fraction in silicosis patients expos ronmental factors for SSc

    H. Hayashi, Y. Miura, M. Maeda, S. Murakami, Y. Nishimura, M. Kusaka, W. Fujimoto, T. Otsuki

    JOURNAL OF INVESTIGATIVE DERMATOLOGY   128   S176 - S176   2008年4月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:NATURE PUBLISHING GROUP  

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  • Immunological changes in mesothelioma patients and their experimental detection. 国際誌

    Maeda Megumi, Miura Yoshie, Nishimura Yasumitsu, Murakami Shuko, Hayashi Hiroaki, Kumagai Naoko, Hatayama Tamayo, Katoh Minako, Miyahara Naomi, Yamamoto Shoko, Fukuoka Kazuya, Kishimoto Takumi, Nakano Takashi, Otsuki Takemi

    Clinical Medicine : Circulafory, Respiratory and Pulmonary   2   11 - 17   2008年3月

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    記述言語:英語  

    200800

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  • 珪肺症例におけるCD4^+25^+分画におけるFas分子発現、アポトーシスの検討

    林 宏明, 前田 恵, 村上 周子, 西村 泰光, 草加 勝康, 大槻 剛巳

    日本衞生學雜誌   63 ( 2 )   448 - 448   2008年3月

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  • アスベスト曝露ヒトポリクローナルリンパ球MT-2株のDNAマイクロアレイによる遺伝子発現解析

    前田 恵, 西村 泰光, 村上 周子, 熊谷 直子, 林 宏明, 三浦 由恵, 大槻 剛巳

    日本衞生學雜誌   63 ( 2 )   446 - 446   2008年3月

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  • 石綿曝露培養後の末梢血NK細胞におけるNKp46発現低下および石綿曝露NK細胞亜株YT-CB5におけるERKリン酸化低下

    西村 泰光, 前田 恵, 村上 周子, 熊谷 直子, 林 宏明, 三浦 由恵, 大槻 剛巳

    日本衞生學雜誌   63 ( 2 )   447 - 447   2008年3月

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  • ヒトポリクローナルT細胞株のアスベスト誘導アポトーシス抵抗性亜株における転写因子に関する検討

    村上 周子, 前田 恵, 三浦 由恵, 西村 泰光, 熊谷 直子, 林 宏明, 大槻 剛巳

    日本衞生學雜誌   63 ( 2 )   547 - 547   2008年3月

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  • Immunological alterations found in mesothelioma patients and supporting experimental evidence. 国際誌

    Yoshie Miura, Yasumitsu Nishimura, Megumi Maeda, Shuko Murakami, Hiroaki Hayashi, Kazuya Fukuoka, Takumi Kishimoto, Takashi Nakano, Takemi Otsuki

    Environmental health and preventive medicine   13 ( 2 )   55 - 9   2008年3月

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    記述言語:英語  

    It is common knowledge that exposure to asbestos causes asbestos-related diseases, such as asbestosis, lung cancer and malignant mesothelioma, not only in people who have had long-term contact with asbestos in their work environment but also in residents living near factories that handle asbestos. Since the summer of 2005, these revelations turned into a large medical problem and caused and social unrest. We have focused on the immunological effects of both asbestos and silica on the human immune system. In this brief review, we introduce immunological alterations found in patients with malignant mesothelioma and describe the experimental background in which these were found. Analyzing the immunological effects of asbestos may improve our understanding of the biological effects of asbestos.

    DOI: 10.1007/s12199-007-0012-y

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  • Immunological alterations found in mesothelioma patients and supporting experimental evidence. 国際誌

    Yoshie Miura, Yasumitsu Nishimura, Megumi Maeda, Shuko Murakami, Hiroaki Hayashi, Kazuya Fukuoka, Takumi Kishimoto, Takashi Nakano, Takemi Otsuki

    Environmental health and preventive medicine   13 ( 2 )   55 - 9   2008年3月

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    記述言語:英語  

    It is common knowledge that exposure to asbestos causes asbestos-related diseases, such as asbestosis, lung cancer and malignant mesothelioma, not only in people who have had long-term contact with asbestos in their work environment but also in residents living near factories that handle asbestos. Since the summer of 2005, these revelations turned into a large medical problem and caused and social unrest. We have focused on the immunological effects of both asbestos and silica on the human immune system. In this brief review, we introduce immunological alterations found in patients with malignant mesothelioma and describe the experimental background in which these were found. Analyzing the immunological effects of asbestos may improve our understanding of the biological effects of asbestos.

    DOI: 10.1007/s12199-007-0012-y

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  • Salt-adaptation of tobacco BY2 cells induces change in glycoform of N-glycans: Enhancement of exo- and endo-glycosidase activities by salt-adaptation

    Yoshinobu Kimura, Takao Watanabe, Mariko Kimura, Megumi Maeda, Yoshiyuki Murata, Kazuhito Fujjyama

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   72 ( 2 )   514 - 522   2008年2月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    In this report, we describe that a salt adaptation of plant cells induces glycoform changes in N-glycoproteins. Intracellular and cell-wall glycopeptides were prepared from glycoproteins expressed in wild-type BY2 cells and salt-adapted cells. N-Glycans were liberated from those glycopeptides by hydrazinolysis, and the released oligosaccharides were N-acetylated and pyridylaminated. The structures of pyridylaminated (PA-) N-glycans were analyzed by a combination of two-dimensional sugar-chain mapping, MS analysis, and exoglycosidase digestion. In both wild-type cells and salt-adapted cells, the plant complex type structure was predominant among N-glycans expressed on glycoproteins, but we found that the Man2Xyl1Fuc1GlcNAc2 structure was significantly expressed on intracellular and cell-wall glycoproteins of the salt-adapted cells. Furthermore, enhancement of the specific activities of alpha-mannosidase and beta-N-acetylglucosaminidase was observed in the salt-adapted BY2 cells, suggesting that the glycoform changes are due to changes in glycosidase activities.

    DOI: 10.1271/bbb.70592

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  • Glycoform analysis of Japanese cypress pollen allergeng cha o 1: A comparison of the glycoforms of cedar and cypress pollen allergens

    Yoshinobu Kimura, Misao Kuroki, Megumi Maeda, Mitsuhiro Okano, Minehiko Yokoyama, Kosuke Kino

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   72 ( 2 )   485 - 491   2008年2月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    A Japanese cypress (Chamaecyparis obtusa) pollen allergen, Cha o 1, is one of the major allergens that cause allergic pollinosis in Japan. Although it has been found that Cha o 1 is glycosylated and that the amino acid sequence is highly homologous with that of Japanese cedar pollen allergen (Cry j 1), the structure of N-glycans linked to Cha o1 remains to be determined. In this study, therefore, we analyzed the structures of the N-glycans of Cha o1. The N-glycans were liberated by hydrazinolysis from purified Cha o 1, and the resulting sugar chains were N-acetylated and pyridylaminated. The structures of pyridylaminated N-glycans were analyzed by a combination of exoglycosidase digestion, two dimensional (2D-) sugar chain mapping, and electrospray ionization mass spectrometry analysis. Structural analysis indicated that the major N-glycan structure of Cha o1 is GIcNAc2Man3Xyl1-Fuc1GlcNAc2 (89%), and that high-mannose type structures (Man9GlcNAc2, Man7GIcNAc2) occur as minor components (11%).

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  • Salt-adaptation of tobacco BY2 cells induces change in glycoform of N-glycans: Enhancement of exo- and endo-glycosidase activities by salt-adaptation 国際誌

    Yoshinobu Kimura, Takao Watanabe, Mariko Kimura, Megumi Maeda, Yoshiyuki Murata, Kazuhito Fujjyama

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   72 ( 2 )   514 - 522   2008年2月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    In this report, we describe that a salt adaptation of plant cells induces glycoform changes in N-glycoproteins. Intracellular and cell-wall glycopeptides were prepared from glycoproteins expressed in wild-type BY2 cells and salt-adapted cells. N-Glycans were liberated from those glycopeptides by hydrazinolysis, and the released oligosaccharides were N-acetylated and pyridylaminated. The structures of pyridylaminated (PA-) N-glycans were analyzed by a combination of two-dimensional sugar-chain mapping, MS analysis, and exoglycosidase digestion. In both wild-type cells and salt-adapted cells, the plant complex type structure was predominant among N-glycans expressed on glycoproteins, but we found that the Man2Xyl1Fuc1GlcNAc2 structure was significantly expressed on intracellular and cell-wall glycoproteins of the salt-adapted cells. Furthermore, enhancement of the specific activities of alpha-mannosidase and beta-N-acetylglucosaminidase was observed in the salt-adapted BY2 cells, suggesting that the glycoform changes are due to changes in glycosidase activities.

    DOI: 10.1271/bbb.70592

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  • Glycoform analysis of Japanese cypress pollen allergeng cha o 1: A comparison of the glycoforms of cedar and cypress pollen allergens 国際誌

    Yoshinobu Kimura, Misao Kuroki, Megumi Maeda, Mitsuhiro Okano, Minehiko Yokoyama, Kosuke Kino

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   72 ( 2 )   485 - 491   2008年2月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    A Japanese cypress (Chamaecyparis obtusa) pollen allergen, Cha o 1, is one of the major allergens that cause allergic pollinosis in Japan. Although it has been found that Cha o 1 is glycosylated and that the amino acid sequence is highly homologous with that of Japanese cedar pollen allergen (Cry j 1), the structure of N-glycans linked to Cha o1 remains to be determined. In this study, therefore, we analyzed the structures of the N-glycans of Cha o1. The N-glycans were liberated by hydrazinolysis from purified Cha o 1, and the resulting sugar chains were N-acetylated and pyridylaminated. The structures of pyridylaminated N-glycans were analyzed by a combination of exoglycosidase digestion, two dimensional (2D-) sugar chain mapping, and electrospray ionization mass spectrometry analysis. Structural analysis indicated that the major N-glycan structure of Cha o1 is GIcNAc2Man3Xyl1-Fuc1GlcNAc2 (89%), and that high-mannose type structures (Man9GlcNAc2, Man7GIcNAc2) occur as minor components (11%).

    DOI: 10.1271/bbb.70572

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  • Lifestyle and T-helper 1 and 2 related cytokines in healthy volunteers

    Miura Yoshie, Ishibashi T, Tatsukawa TT, Maeda Megumi, Murakami Shuko, Nishimura Yasumitsu, Kumagai Naoko, Hayashi Hiroaki, Chen Ying, Hyodo Fuminori, Kojima S, Fujii M, Morimoto K

    Kawasaki Medical Journal   34 ( 3 )   171 - 177   2008年

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    記述言語:英語   出版者・発行元:川崎医科大学  

    200800

    DOI: 10.11482/2008/KMJ34(3)171-177.2008.pdf

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    その他リンク: http://id.nii.ac.jp/1162/00001488/

  • P-334 アスベスト曝露症例末梢血CD3+T細胞におけるT細胞受容体Vβ発現の検討と実験系における検証(一般演題(ポスター)35 免疫・サイトカイン,第48回日本肺癌学会総会号)

    大槻 剛巳, 前田 恵, 村上 周子, 林 宏明, 福岡 和也, 栗林 康造, 岸本 卓巳, 草加 勝康, 中野 孝司, 西村 泰光

    肺癌   47 ( 5 )   608 - 608   2007年10月

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    記述言語:日本語   出版者・発行元:日本肺癌学会  

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  • P-333 悪性中皮腫患者NK細胞の細胞傷害性及びNK活性化レセプターNKp46発現量の低下(一般演題(ポスター)35 免疫・サイトカイン,第48回日本肺癌学会総会号)

    西村 泰光, 前田 恵, 村上 周子, 林 宏明, 三浦 由恵, 福岡 和也, 中野 孝司, 大槻 剛巳

    肺癌   47 ( 5 )   608 - 608   2007年10月

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    記述言語:日本語   出版者・発行元:日本肺癌学会  

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  • WS11-1 悪性中皮腫症例末梢血CD4+T細胞におけるBcl-2高発現と実験的検証(ワークショップ 悪性中皮腫,第48回日本肺癌学会総会号)

    大槻 剛巳, 三浦 由恵, 前田 恵, 林 宏明, 村上 周子, 福岡 和也, 栗林 康造, 岸本 卓巳, 中野 孝司, 西村 泰光

    肺癌   47 ( 5 )   467 - 467   2007年10月

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    記述言語:日本語   出版者・発行元:日本肺癌学会  

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  • Immunological effects of silica and asbestos. 国際誌

    Takemi Otsuki, Megumi Maeda, Shuko Murakami, Hiroaki Hayashi, Yoshie Miura, Masayasu Kusaka, Takashi Nakano, Kazuya Fukuoka, Takumi Kishimoto, Fuminori Hyodoh, Ayako Ueki, Yasumitsu Nishimura

    Cellular & molecular immunology   4 ( 4 )   261 - 8   2007年8月

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    記述言語:英語  

    Silicosis patients (SILs) and patients who have been exposed to asbestos develop not only respiratory diseases but also certain immunological disorders. In particular, SIL sometimes complicates autoimmune diseases such as systemic scleroderma, rheumatoid arthritis (known as Caplan syndrome), and systemic lupus erythematoses. In addition, malignant complications such as lung cancer and malignant mesothelioma often occur in patients exposed to asbestos, and may be involved in the reduction of tumor immunity. Although silica-induced disorders of autoimmunity have been explained as adjuvant-type effects of silica, more precise analyses are needed and should reflect the recent progress in immunomolecular findings. A brief summary of our investigations related to the immunological effects of silica/asbestos is presented. Recent advances in immunomolecular studies led to detailed analyses of the immunological effects of asbestos and silica. Both affect immuno-competent cells and these effects may be associated with the pathophysiological development of complications in silicosis and asbestos-exposed patients such as the occurrence of autoimmune disorders and malignant tumors, respectively. In addition, immunological analyses may lead to the development of new clinical tools for the modification of the pathophysiological aspects of diseases such as the regulation of autoimmunity or tumor immunity using cell-mediated therapies, various cytokines, and molecule-targeting therapies. In particular, as the incidence of asbestos-related malignancies is increasing and such malignancies have been a medical and social problem since the summer of 2005 in Japan, efforts should be focused on developing a cure for these diseases to eliminate nationwide anxiety.

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  • Immunological effects of silica and asbestos. 国際誌

    Takemi Otsuki, Megumi Maeda, Shuko Murakami, Hiroaki Hayashi, Yoshie Miura, Masayasu Kusaka, Takashi Nakano, Kazuya Fukuoka, Takumi Kishimoto, Fuminori Hyodoh, Ayako Ueki, Yasumitsu Nishimura

    Cellular & molecular immunology   4 ( 4 )   261 - 8   2007年8月

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    記述言語:英語  

    Silicosis patients (SILs) and patients who have been exposed to asbestos develop not only respiratory diseases but also certain immunological disorders. In particular, SIL sometimes complicates autoimmune diseases such as systemic scleroderma, rheumatoid arthritis (known as Caplan syndrome), and systemic lupus erythematoses. In addition, malignant complications such as lung cancer and malignant mesothelioma often occur in patients exposed to asbestos, and may be involved in the reduction of tumor immunity. Although silica-induced disorders of autoimmunity have been explained as adjuvant-type effects of silica, more precise analyses are needed and should reflect the recent progress in immunomolecular findings. A brief summary of our investigations related to the immunological effects of silica/asbestos is presented. Recent advances in immunomolecular studies led to detailed analyses of the immunological effects of asbestos and silica. Both affect immuno-competent cells and these effects may be associated with the pathophysiological development of complications in silicosis and asbestos-exposed patients such as the occurrence of autoimmune disorders and malignant tumors, respectively. In addition, immunological analyses may lead to the development of new clinical tools for the modification of the pathophysiological aspects of diseases such as the regulation of autoimmunity or tumor immunity using cell-mediated therapies, various cytokines, and molecule-targeting therapies. In particular, as the incidence of asbestos-related malignancies is increasing and such malignancies have been a medical and social problem since the summer of 2005 in Japan, efforts should be focused on developing a cure for these diseases to eliminate nationwide anxiety.

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  • Free oligosaccharides in the cytosol of Caenorhabditis elegans are generated through endoplasmic reticulum-Golgi trafficking 国際誌

    Toshihiko Kato, Kumiko Kitamura, Megumi Maeda, Yoshinobu Kimura, Takane Katayama, Hisashi Ashida, Kenji Yamamoto

    JOURNAL OF BIOLOGICAL CHEMISTRY   282 ( 30 )   22080 - 22088   2007年7月

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    記述言語:英語   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Free oligosaccharides (FOSs) in the cytosol of eukaryotic cells are mainly generated during endoplasmic reticulum (ER)-associated degradation (ERAD) of misfolded glycoproteins. We analyzed FOS of the nematode Caenorhabditis elegans to elucidate its detailed degradation pathway. The major FOSs were high mannose-type ones bearing 3-9 Man residues. About 94% of the total FOSs had one GlcNAc at their reducing end (FOS-GN1), and the remaining 6% had two GlcNAc (FOS-GN2). A cytosolic endo-beta-N-acetylglucosaminidase mutant (tm1208) accumulated FOS-GN2, indicating involvement of the enzyme in conversion of FOS-GN2 into FOS-GN1. The most abundant FOS in the wild type was Man(5)GlcNAc(1), the M5A' isomer (Man alpha 1- 3(Man alpha 1-6)Man alpha 1-6(Man alpha 1-3)Man beta 1-4GlcNAc), which is different from the corresponding M5B' (Man alpha 1-2Man alpha 1- 2Man alpha 1-3(Man alpha 1-6)Man beta 1-4GlcNAc) in mammals. Analyses of FOS in worms treated with Golgi alpha-mannosidase I inhibitors revealed decreases in Man(5)GlcNAc(1) and increases in Man(7)GlcNAc(1). These results suggested that Golgi alpha-mannosidase I-like enzyme is involved in the production of Man(5-6)- GlcNAc(1), which is unlike in mammals, in which cytosolic alpha-mannosidase is involved. Thus, we assumed that major FOSs in C. elegans were generated through Golgi trafficking. Analysis of FOSs from a Golgi alpha-mannosidase II mutant (tm1078) supported this idea, because GlcNAc(1)Man(5)GlcNAc(1), which is formed by the Golgi-resident GlcNAc-transferase I, was found as a FOS in the mutant. We concluded that significant amounts of misfolded glycoproteins in C. elegans are trafficked to the Golgi and are directly or indirectly retro-translocated into the cytosol to be degraded.

    DOI: 10.1074/jbc.M700805200

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  • Keynote lecture in the 13th Japanese Society of Immunotoxicology (JSIT 2006) : -Pathophysiological Development and Immunotoxicology: what we have found from research related to silica and silicate such as asbestos-. 国際誌

    Takemi Otsuki, Yoshie Miura, Megumi Maeda, Hiroaki Hayashi, Shuko Murakami, Maolong Dong, Yasumitsu Nishimura

    Environmental health and preventive medicine   12 ( 4 )   153 - 60   2007年7月

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    記述言語:英語  

    Silica and silicates may disturb immune functions such as autoimmunity and tumor immunity, because people who are exposed to the materials sometimes develop autoimmune and malignant diseases, respectively. Although silica-induced disorders of autoimmunity have been explained as adjuvant-type effects of silica, more precise analyses are needed and should reflect the recent progress in immunomolecular findings. A brief summary of our investigations related to the immunological effects of silica/asbestos is presented. Recent advances in immunomolecular studies led to detailed analyses of the immunological effects of asbestos and silica. Both affect immuno-competent cells and these effects may be associated with the pathophysiological development of complications in silicosis and asbestos-exposed patients such as the occurrence of autoimmune disorders and malignant tumors, respectively. In addition, immunological analyses may lead to the development of new clinical tools for the modification of the pathophysiological aspects of diseases such as the regulation of autoimmunity or tumor immunity using cell-mediated therapies, various cytokines, and molecule-targeting therapies. In particular, as the incidence of asbestos-related malignancies is increasing and such malignancies have been a medical and social problem since the summer in 2005 in Japan, efforts should be focused on developing a cure for these diseases to eliminate the nation wide anxiety about these malignancies.

    DOI: 10.1007/BF02897984

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  • Keynote lecture in the 13th Japanese Society of Immunotoxicology (JSIT 2006) : -Pathophysiological Development and Immunotoxicology: what we have found from research related to silica and silicate such as asbestos-. 国際誌

    Takemi Otsuki, Yoshie Miura, Megumi Maeda, Hiroaki Hayashi, Shuko Murakami, Maolong Dong, Yasumitsu Nishimura

    Environmental health and preventive medicine   12 ( 4 )   153 - 60   2007年7月

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    記述言語:英語  

    Silica and silicates may disturb immune functions such as autoimmunity and tumor immunity, because people who are exposed to the materials sometimes develop autoimmune and malignant diseases, respectively. Although silica-induced disorders of autoimmunity have been explained as adjuvant-type effects of silica, more precise analyses are needed and should reflect the recent progress in immunomolecular findings. A brief summary of our investigations related to the immunological effects of silica/asbestos is presented. Recent advances in immunomolecular studies led to detailed analyses of the immunological effects of asbestos and silica. Both affect immuno-competent cells and these effects may be associated with the pathophysiological development of complications in silicosis and asbestos-exposed patients such as the occurrence of autoimmune disorders and malignant tumors, respectively. In addition, immunological analyses may lead to the development of new clinical tools for the modification of the pathophysiological aspects of diseases such as the regulation of autoimmunity or tumor immunity using cell-mediated therapies, various cytokines, and molecule-targeting therapies. In particular, as the incidence of asbestos-related malignancies is increasing and such malignancies have been a medical and social problem since the summer in 2005 in Japan, efforts should be focused on developing a cure for these diseases to eliminate the nation wide anxiety about these malignancies.

    DOI: 10.1007/BF02897984

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  • Free oligosaccharides in the cytosol of Caenorhabditis elegans are generated through endoplasmic reticulum-Golgi trafficking

    Toshihiko Kato, Kumiko Kitamura, Megumi Maeda, Yoshinobu Kimura, Takane Katayama, Hisashi Ashida, Kenji Yamamoto

    JOURNAL OF BIOLOGICAL CHEMISTRY   282 ( 30 )   22080 - 22088   2007年7月

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    記述言語:英語   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Free oligosaccharides (FOSs) in the cytosol of eukaryotic cells are mainly generated during endoplasmic reticulum (ER)-associated degradation (ERAD) of misfolded glycoproteins. We analyzed FOS of the nematode Caenorhabditis elegans to elucidate its detailed degradation pathway. The major FOSs were high mannose-type ones bearing 3-9 Man residues. About 94% of the total FOSs had one GlcNAc at their reducing end (FOS-GN1), and the remaining 6% had two GlcNAc (FOS-GN2). A cytosolic endo-beta-N-acetylglucosaminidase mutant (tm1208) accumulated FOS-GN2, indicating involvement of the enzyme in conversion of FOS-GN2 into FOS-GN1. The most abundant FOS in the wild type was Man(5)GlcNAc(1), the M5A' isomer (Man alpha 1- 3(Man alpha 1-6)Man alpha 1-6(Man alpha 1-3)Man beta 1-4GlcNAc), which is different from the corresponding M5B' (Man alpha 1-2Man alpha 1- 2Man alpha 1-3(Man alpha 1-6)Man beta 1-4GlcNAc) in mammals. Analyses of FOS in worms treated with Golgi alpha-mannosidase I inhibitors revealed decreases in Man(5)GlcNAc(1) and increases in Man(7)GlcNAc(1). These results suggested that Golgi alpha-mannosidase I-like enzyme is involved in the production of Man(5-6)- GlcNAc(1), which is unlike in mammals, in which cytosolic alpha-mannosidase is involved. Thus, we assumed that major FOSs in C. elegans were generated through Golgi trafficking. Analysis of FOSs from a Golgi alpha-mannosidase II mutant (tm1078) supported this idea, because GlcNAc(1)Man(5)GlcNAc(1), which is formed by the Golgi-resident GlcNAc-transferase I, was found as a FOS in the mutant. We concluded that significant amounts of misfolded glycoproteins in C. elegans are trafficked to the Golgi and are directly or indirectly retro-translocated into the cytosol to be degraded.

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  • 【アスベスト問題から得る教訓】 アスベストの生体影響に関する研究について

    大槻剛巳, 前田恵, 林宏明, 董茂龍, 三浦由恵, 西村泰光

    安全医学 (Journal of Medical Safety)   3 ( 2 )   92 - 103   2007年5月

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    記述言語:日本語   出版者・発行元:日本予防医学リスクマネージメント学会  

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    その他リンク: http://search.jamas.or.jp/link/ui/2007331578

  • 珪酸・アスベストの免疫系への影響

    大槻剛巳, 前田恵, 三浦由恵, 兵藤文則, 植木絢子, 村上周子, 林宏明, 勝山博信, 富田正文, 西村泰光

    日本職業災害医学会会誌   55 ( 3 )   113 - 120   2007年5月

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  • 科学技術振興研究発表AS-1 珪酸・アスベストの免疫系への影響—Research on the immunological effects of silica and asbestos 査読

    大槻 剛巳, 前田 恵, 三浦 由恵

    日本職業・災害医学会会誌 = Japanese journal of occupational medicine and traumatology   55 ( 3 )   113 - 120   2007年5月

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    記述言語:日本語   出版者・発行元:日本職業・災害医学会  

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    その他リンク: https://search.jamas.or.jp/link/ui/2008052691

  • アスベスト低濃度・長期曝露によるヒトポリクローナルリンパ球MT-2株の抑制性サイトカイン産生誘導

    前田 恵, 三浦 由恵, 西村 泰光, 林 宏明, 董 茂龍, 上坂 亜由子, 栗林 康造, 中野 孝司, 岸本 卓巳, 大槻 剛巳

    日本衞生學雜誌   62 ( 2 )   607 - 607   2007年3月

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  • 珪肺症例におけるCD4+25+分画における慢性活性化T細胞検討

    林 宏明, 三浦 由恵, 前田 恵, 兵藤 文則, 西村 泰光, 勝山 博信, 草加 勝康, 大槻 剛巳

    日本衞生學雜誌   62 ( 2 )   609 - 609   2007年3月

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  • アスベスト長期曝露によるNK細胞の機能変化

    西村 泰光, 三浦 由恵, 前田 恵, 林 宏明, 大槻 剛巳

    日本衞生學雜誌   62 ( 2 )   404 - 407   2007年3月

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    記述言語:日本語   出版者・発行元:日本衛生学会  

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    その他リンク: http://search.jamas.or.jp/link/ui/2007151909

  • 石綿長期曝露後のヒトNK細胞株YT-A1におけるNK活性化レセプター2B4の機能低下

    西村 泰光, 三浦 由恵, 前田 恵, 林 宏明, 大槻 剛巳

    日本衞生學雜誌   62 ( 2 )   610 - 610   2007年3月

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  • アスベストに関連する発がん機構と免疫学的影響

    大槻剛巳, 三浦由恵, 林宏明, 前田恵, 董茂龍, 西村泰光, 富田正文, 勝山博信

    Biomedical Research on Trace Elements   17 ( 4 )   385 - 390   2006年12月

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    記述言語:日本語   出版者・発行元:日本微量元素学会  

    With the increasing detection of patients with malignant mesothelioma among residents living near factories and facilities handling asbestos in the past, asbestos-related diseases have become a big medical, social and political issue in Japan. Therefore, we should seriously consider what we, as researchers into the biological effects of asbestos in the field of preventive medicine, can do to clarify the problem and which type of investigations may eliminate the anxiety of Japanese people. One answer is to establish biomarkers for asbestos exposure, since most people do not know for sure whether or not they were exposed to asbestos 30 to 40 years ago. In addition, we should develop markers to detect asbestos-induced malignancies such as lung cancer and malignant mesothelioma. Moreover, the establishment of the markers for molecular prevention of asbestos-induced carcinogenesis would be much appreciated news to persons feeling anxiety about the uncertainty of past exposure to asbestos.

    DOI: 10.11299/brte.17.385

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  • TRAIL expression up-regulated by interferon-γ via phosphrylation of STAT1 induces myeloma cell death.

    Miura Yoshie, Tsujioka Takayuki, Nishimura Yasumitsu, Sakaguchi Haruko, Maeda Megumi, Hayashi Hiroaki, Dong Maolong, Hyodoh Fuminori, Yata Ken-ichiro, Wada Hideho, Sugihara Takashi, Otsuki Takemi

    Anticancer research   26 ( 6B )   4115 - 4124   2006年11月

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  • TRAIL expression up-regulated by interferon-gamma via phosphorylation of STAT1 induces myeloma cell death

    Yoshie Miura, Takayuki Tsujioka, Yasumitsu Nishimura, Haruko Sakaguchi, Megumi Maeda, Hiroaki Hayashi, Maolong Dong, Fuminori Hyodoh, Ken-Ichiro Yata, Hideho Wada, Takashi Sugihara, Takemi Otsuki

    ANTICANCER RESEARCH   26 ( 6B )   4115 - 4124   2006年11月

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    記述言語:英語   出版者・発行元:INT INST ANTICANCER RESEARCH  

    Background: Although the cellular and molecular biological effects of interferon (IFN)-alpha have been well-investigated, the effects of IFN-gamma are less understood. Materials and Methods: Eleven human myeloma cell lines with various myeloma-specific chromosomal translocations and overexpression of oncogenes were cultured with 1,000 U/ml of IFN-gamma. In the KMS-20 cells, which showed growth inhibition due to IFN-gamma, trail expression, status of the Janus kinase (JAK)/STAT pathway were analyzed. Results: KHS-20 cells showed marked up-regulation of trail, activation of STAT1 and TRAIL hyperproduction induced by IFN-gamma. Conclusion: The effects of IFN-gamma on growth inhibition of KMS-20 cells were characterized by activation of the JAK/STAT signalling pathway, particularly STAT1 phosphorylation, enhanced secretion of TRAIL, and auto/paracrine usage of secreted TRAIL to induce apoptotic cell death. From these results, IFN-gamma may be considered one of the drugs to be used in future multidrug chemotherapeutic regimens for myeloma patients.

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  • Tumor antigen occurs in N-glycan of royal jelly glycoproteins: Honeybee cells synthesize T-antigen unit in N-glycan moiety 国際誌

    Yoshinobu Kimura, Takayuki Ushijima, Megumi Maeda, Yoichiro Hama, Maniko Kimura, Kiyoshi Okihara, Hiroyuki Sugimoto, Hideo Yamada

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   70 ( 10 )   2583 - 2587   2006年10月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    In our previous paper (Kimura, Y., et al, Biosci. Biotechnol. Biochem., 67, 1852-1856, 2003), we found that a complex type N-glycans containing beta 1-3 galactose residue occurs on royal jelly glycoproteins. During structural analysis of minor components of royal jelly N-glycans, we found complex type N-glycans bearing both galactose and N-acetylgalactosamine residues. Detailed structural analysis of pyridylaminated oligosaccharide revealed that the newly found N-glycan had a complex type structure harboring a tumor marker (T-antigen) unit: Gal beta 1-3GaINAc beta 1-4GlcNAc beta 1-2Man alpha 1-6 (Gal beta 1-3GalNAc beta 1-4GlcNAc beta 1-2Man alpha 1-3) Man beta 1-4GlcNAc beta 1-4GIcNAc. To our knowledge, this may be the first report of the presence of the T-antigen unit in the N-glycan moiety of eucaryotic glycoproteins.

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  • Involvement of IL-10 and Bcl-2 in resistance against an asbestos-induced apoptosis of T cells. 国際誌

    Yoshie Miura, Yasumitsu Nishimura, Hironobu Katsuyama, Megumi Maeda, Hiroaki Hayashi, Maolong Dong, Fuminori Hyodoh, Masafumi Tomita, Yoshinobu Matsuo, Ayuko Uesaka, Kozo Kuribayashi, Takashi Nakano, Takumi Kishimoto, Takemi Otsuki

    Apoptosis : an international journal on programmed cell death   11 ( 10 )   1825 - 35   2006年10月

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    To analyze the possibility that immunological alteration in asbestos-related diseases (ARDs) such as asbestosis (ASB) and malignant mesothelioma (MM) may affect the progression of cancers, a human adult T cell leukemia virus-immortalized T cell line (MT-2Org) was continuously exposed to 10 mug/ml of chrysotile-B (CB), an asbestos. After at least 8 months of exposure, the rate of apoptosis in the cells became very low and the resultant subline was designated MT-2Rst. The MT-2Rst cells were characterized by (i) enhanced expression of bcl-2, with regain of apoptosis-sensitivity by reduction of bcl-2 by siRNA, (ii) excess IL-10 secretion and expression, and (iii) activation of STAT3 that was inhibited by PP2, a specific inhibitor of Src family kinases. These results suggested that the contact between cells and asbestos may affect the human immune system and trigger a cascade of biological events such as activation of Src family kinases, enhancement of IL-10 expression, STAT3 activation and Bcl-2 overexpression. This speculation was partially confirmed by the detection of elevated bcl-2 expression levels in CD4 + peripheral blood T cells from patients with MM compared with those from patients with ASB or healthy donors. Further studies will be required to verify the role of T cells with enhanced bcl-2 expression in tumor progression induced by asbestos exposure.

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  • Tumor antigen occurs in N-glycan of royal jelly glycoproteins: Honeybee cells synthesize T-antigen unit in N-glycan moiety

    Yoshinobu Kimura, Takayuki Ushijima, Megumi Maeda, Yoichiro Hama, Maniko Kimura, Kiyoshi Okihara, Hiroyuki Sugimoto, Hideo Yamada

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   70 ( 10 )   2583 - 2587   2006年10月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    In our previous paper (Kimura, Y., et al, Biosci. Biotechnol. Biochem., 67, 1852-1856, 2003), we found that a complex type N-glycans containing beta 1-3 galactose residue occurs on royal jelly glycoproteins. During structural analysis of minor components of royal jelly N-glycans, we found complex type N-glycans bearing both galactose and N-acetylgalactosamine residues. Detailed structural analysis of pyridylaminated oligosaccharide revealed that the newly found N-glycan had a complex type structure harboring a tumor marker (T-antigen) unit: Gal beta 1-3GaINAc beta 1-4GlcNAc beta 1-2Man alpha 1-6 (Gal beta 1-3GalNAc beta 1-4GlcNAc beta 1-2Man alpha 1-3) Man beta 1-4GlcNAc beta 1-4GIcNAc. To our knowledge, this may be the first report of the presence of the T-antigen unit in the N-glycan moiety of eucaryotic glycoproteins.

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  • Involvement of IL-10 and Bcl-2 in resistance against an asbestos-induced apoptosis of T cells. 国際誌

    Yoshie Miura, Yasumitsu Nishimura, Hironobu Katsuyama, Megumi Maeda, Hiroaki Hayashi, Maolong Dong, Fuminori Hyodoh, Masafumi Tomita, Yoshinobu Matsuo, Ayuko Uesaka, Kozo Kuribayashi, Takashi Nakano, Takumi Kishimoto, Takemi Otsuki

    Apoptosis : an international journal on programmed cell death   11 ( 10 )   1825 - 35   2006年10月

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    記述言語:英語  

    To analyze the possibility that immunological alteration in asbestos-related diseases (ARDs) such as asbestosis (ASB) and malignant mesothelioma (MM) may affect the progression of cancers, a human adult T cell leukemia virus-immortalized T cell line (MT-2Org) was continuously exposed to 10 mug/ml of chrysotile-B (CB), an asbestos. After at least 8 months of exposure, the rate of apoptosis in the cells became very low and the resultant subline was designated MT-2Rst. The MT-2Rst cells were characterized by (i) enhanced expression of bcl-2, with regain of apoptosis-sensitivity by reduction of bcl-2 by siRNA, (ii) excess IL-10 secretion and expression, and (iii) activation of STAT3 that was inhibited by PP2, a specific inhibitor of Src family kinases. These results suggested that the contact between cells and asbestos may affect the human immune system and trigger a cascade of biological events such as activation of Src family kinases, enhancement of IL-10 expression, STAT3 activation and Bcl-2 overexpression. This speculation was partially confirmed by the detection of elevated bcl-2 expression levels in CD4 + peripheral blood T cells from patients with MM compared with those from patients with ASB or healthy donors. Further studies will be required to verify the role of T cells with enhanced bcl-2 expression in tumor progression induced by asbestos exposure.

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  • E prostanoid 2 (EP2)/EP4-mediated suppression of antigen-specific human T-cell responses by prostaglandin E-2 国際誌

    M Okano, Y Sugata, T Fujiwara, R Matsumoto, M Nishibori, K Shimizu, M Maeda, Y Kimura, S Kariya, H Hattori, M Yokoyama, K Kino, K Nishizaki

    IMMUNOLOGY   118 ( 3 )   343 - 352   2006年7月

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    記述言語:英語   出版者・発行元:BLACKWELL PUBLISHING  

    Prostaglandin E-2 (PGE(2)) is a lipid mediator that displays important immunomodulatory properties, such as polarization of cytokine production by T cells. Recent investigations have revealed that the effect of PGE(2) on cytokine production is greatly influenced by external stimuli; however, it is unclear whether PGE(2) plays a significant role in major histocompatibility complex-mediated antigen-specific T-cell responses via binding to one of four subtypes of E prostanoid (EP) receptor alone or in combination. In the present study, we sought to determine the effect of PGE(2) on antigen-specific CD4(+) T-cell responses in humans, especially in terms of receptor specificity. We used purified protein derivative (PPD) and Cry j 1 as T helper type 1 (Th1) and Th2-inducing antigens, respectively. We generated several different Cry j 1- and PPD-specific T-cell lines (TCLs). PGE(2) significantly and dose-dependently inhibited the proliferation and subsequent production of interleukin-4 by Cry j 1-specific TCLs and of interferon-gamma by PPD-specific TCLs upon antigen stimulation. Administration of EP2 receptor agonist and EP4 receptor agonist suppressed these responses in an adenylate cyclase-dependent manner, while EP1 and EP3 receptor agonists did not. Messenger RNA for EP2, EP3 and EP4, but not EP1, receptors were detected in Cry j 1- and PPD-specific TCLs, and no differences in EP receptor expression were observed between them. Furthermore, PGE(2) and EP2 receptor agonist significantly inhibited interleukin-5 and interferon-gamma production by peripheral blood mononuclear cells in response to Cry j 1 and PPD stimulation, respectively. These results suggest that PGE(2) suppresses both Th1- and Th2-polarized antigen-specific human T-cell responses via a cAMP-dependent EP2/EP4-mediated pathway.

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  • E prostanoid 2 (EP2)/EP4-mediated suppression of antigen-specific human T-cell responses by prostaglandin E-2

    M Okano, Y Sugata, T Fujiwara, R Matsumoto, M Nishibori, K Shimizu, M Maeda, Y Kimura, S Kariya, H Hattori, M Yokoyama, K Kino, K Nishizaki

    IMMUNOLOGY   118 ( 3 )   343 - 352   2006年7月

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    記述言語:英語   出版者・発行元:BLACKWELL PUBLISHING  

    Prostaglandin E-2 (PGE(2)) is a lipid mediator that displays important immunomodulatory properties, such as polarization of cytokine production by T cells. Recent investigations have revealed that the effect of PGE(2) on cytokine production is greatly influenced by external stimuli; however, it is unclear whether PGE(2) plays a significant role in major histocompatibility complex-mediated antigen-specific T-cell responses via binding to one of four subtypes of E prostanoid (EP) receptor alone or in combination. In the present study, we sought to determine the effect of PGE(2) on antigen-specific CD4(+) T-cell responses in humans, especially in terms of receptor specificity. We used purified protein derivative (PPD) and Cry j 1 as T helper type 1 (Th1) and Th2-inducing antigens, respectively. We generated several different Cry j 1- and PPD-specific T-cell lines (TCLs). PGE(2) significantly and dose-dependently inhibited the proliferation and subsequent production of interleukin-4 by Cry j 1-specific TCLs and of interferon-gamma by PPD-specific TCLs upon antigen stimulation. Administration of EP2 receptor agonist and EP4 receptor agonist suppressed these responses in an adenylate cyclase-dependent manner, while EP1 and EP3 receptor agonists did not. Messenger RNA for EP2, EP3 and EP4, but not EP1, receptors were detected in Cry j 1- and PPD-specific TCLs, and no differences in EP receptor expression were observed between them. Furthermore, PGE(2) and EP2 receptor agonist significantly inhibited interleukin-5 and interferon-gamma production by peripheral blood mononuclear cells in response to Cry j 1 and PPD stimulation, respectively. These results suggest that PGE(2) suppresses both Th1- and Th2-polarized antigen-specific human T-cell responses via a cAMP-dependent EP2/EP4-mediated pathway.

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  • アスベストの物理化学的諸性質と生体影響

    大槻剛巳, 三浦由恵, 林宏明, 前田恵, 董茂龍, 西村泰光

    The Lung perspectives   14 ( 3 )   311 - 315   2006年7月

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  • Glycoform analysis of N-glycans linked to glycoproteins expressed in rice culture cells: Predominant occurrence of complex type N-glycans

    Megumi Maeda, Yoshinobu Kimura

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   70 ( 6 )   1356 - 1363   2006年6月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    Although it has been found that plant endo-beta-N-acetylglucosaminidase shows strong activity towards denatured glycoproteins and glycopeptides with high-mannose type N-glycans and free high-mannose type N-glycans bearing the chitobiosyl unit, the endogenous substrates for plant endoglycosidase have not yet been identified. Recently we purified and characterized an endo-beta-N-acetylglucosaminidase from rice culture cells and identified the gene encoded (Maeda, M., and Kimura, Y., Trends Glycosci. Glycotech., 17, 205-214 (2005)). Furthermore, we found structural features of free N-glycans in the cells, indicating that high-mannose type species (Man(9.5)GlcNAc(1)) occur at concentration of several micromolar (mu m). Hence, in this study we analyzed glycoform of N-glycans linked to glycoproteins expressed in rice culture cells to see whether endogenous glycoproteinous substrate occurs in reasonable amounts. Structural analysis revealed that more than 95% of total N-glycans linked to glycoproteins in the rice cells had the plant complex type structure, including Lewis a epitope-harboring type, although high-mannose type structures account for less than 5% of total N-glycans.

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  • Glycoform analysis of N-glycans linked to glycoproteins expressed in rice culture cells: Predominant occurrence of complex type N-glycans 国際誌

    Megumi Maeda, Yoshinobu Kimura

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   70 ( 6 )   1356 - 1363   2006年6月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    Although it has been found that plant endo-beta-N-acetylglucosaminidase shows strong activity towards denatured glycoproteins and glycopeptides with high-mannose type N-glycans and free high-mannose type N-glycans bearing the chitobiosyl unit, the endogenous substrates for plant endoglycosidase have not yet been identified. Recently we purified and characterized an endo-beta-N-acetylglucosaminidase from rice culture cells and identified the gene encoded (Maeda, M., and Kimura, Y., Trends Glycosci. Glycotech., 17, 205-214 (2005)). Furthermore, we found structural features of free N-glycans in the cells, indicating that high-mannose type species (Man(9.5)GlcNAc(1)) occur at concentration of several micromolar (mu m). Hence, in this study we analyzed glycoform of N-glycans linked to glycoproteins expressed in rice culture cells to see whether endogenous glycoproteinous substrate occurs in reasonable amounts. Structural analysis revealed that more than 95% of total N-glycans linked to glycoproteins in the rice cells had the plant complex type structure, including Lewis a epitope-harboring type, although high-mannose type structures account for less than 5% of total N-glycans.

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  • Expression of the T cell receptor Vβ repertoire in a human T cell resistant to asbestos-induced apoptosis and peripheral blood T cells from patients with silica and asbestos-related diseases.

    Nishimura Yasumitsu, Miura Yoshie, Maeda Megumi, Hayashi Hiroaki, Dong Maolong, Katsuyama Hironobu, Tomita Masafumi, Hyodoh Fuminori, Uesaka Ayuko, Kuribayashi Kozo, Fukuoka K, Nakano Takashi, Kishimoto Takemi, Otsuki Takemi

    International Journal of Immunopathology and Pharmacology   19 ( 4 )   795 - 805   2006年

  • Expression of the T cell receptor Vβ repertoire in a human T cell resistant to asbestos-induced apoptosis and peripheral blood T cells from patients with silica and asbestos-related diseases

    Y. Nishimura, Y. Miura, M. Maeda, H. Hayashi, M. Dong, H. Katsuyama, M. Tomita, F. Hyodoh, M. Kusaka, A. Uesaka, K. Kuribayashi, K. Fukuoka, T. Nakano, T. Kishimoto, Takemi Otsuki

    International Journal of Immunopathology and Pharmacology   19 ( 4 )   795 - 805   2006年

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    To explore the effects of asbestos and silica on the human immune system, an experimental model of low-dose and long-term exposure was established using a human HTLV-1-immortalized polyclonal T cell line, MT-2 (MT-2Org). MT-2 cells were continuously exposed to asbestos at a concentration (10 μg/ml) which does not induce complete cell death during short-term exposure. After acquiring resistance to CB-induced apoptosis (designated MT-2Rst), an immunological comparison was made between the MT-2Org and MT-2Rst lines in terms of T cell receptor-Vβ (TcR-Vβ) expression. MT-2Rst cells showed excess expression of various TcR-Vβ, although TcR-Vβ-overpresenting cells were characterized as undergoing apoptosis due to first contact with CB. Patients with asbestos-related diseases (ARD), such as asbestosis and malignant mesothelioma, were compared with silicosis (SIL) patients as a disease control and with healthy donors (HD). SIL and ARD not only differed in their causative materials, silica and asbestos as mineral silicates, but also in terms of complications; autoimmune disorders in SIL and tumors in ARD. ARD patients showed a restricted overpresentation of TcR-Vβ without clonal expansion, whereas SIL patients revealed significant overpresentation of TcR-Vβ 7.2. These experimental and clinical analyses indicate the superantigenic and dysregulation of autoimmunity-inducing effects of asbestos and silica, respectively. Copyright © by Biolife, s.a.s.

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  • N-glycan metabolism and plant cell differentiation and growth

    M Megumi, K Yoshinobu

    TRENDS IN GLYCOSCIENCE AND GLYCOTECHNOLOGY   17 ( 97 )   205 - 214   2005年9月

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    記述言語:英語   掲載種別:書評論文,書評,文献紹介等   出版者・発行元:GAKUSHIN PUBL CO  

    Free N-glycans are present at micromolar concentrations in plant cells during their differentiation, growth and maturation stages, and might play a role in processes such as seed germination and fruit ripening. The structure of free N-glycans, which are found in hypocotyls and developing seeds and fruit, can be classified into two types: a high-mannose type (HMT) and a plant complex type (CT); the former, in most cases, has only one GIcNAc residue, while the latter has a chitobiose unit. It is thought that the enzyme endo-o-N-acetylglucosaminidase (endo-pGlcNAc-ase) is involved in the production of HMT sugar chains, whereas the enzyme peptide:N-glycanase (PNGase) is involved in the production of plant CT sugar chains. However, the mechanism and significance of free N-glycan production in plant cells remain obscure. To characterize N-glycan metabolism and the physiological function of free sugar chains, we have investigated the substrate specificities, intracellular distributions, and gene structures of endoP-GlcNAc-ase, PNGase, and alpha-mannosidase in various plants. Here, we report our discovery that endo-beta-GlcNAcase activity begins to increase at a specific stage of tomato ripening, and that the amount of free N-glycans dramatically increases in conjunction with this event. In addition, the structural properties of free N-glycans also change notably as the fruit ripens. This review describes N-glycan metabolism in plant cells, and proposes a role for free sugar chains in the differentiation and growth of plants. The recent finding that plant CT sugar chains are immunoactive is also discussed.

    DOI: 10.4052/tigg.17.205

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  • Glycoform analysis of Japanese cedar pollen allergen, Cry j 1

    M Maeda, M Kamamoto, K Hino, S Yamamoto, M Kimura, M Okano, Y Kimura

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   69 ( 9 )   1700 - 1705   2005年9月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    In our previous study (Y. Kimura et al., Biosci. Biotechnol, Biochem., 69, 137-144 (2005)), we found that plant complex type N-glycans harboring Lewis a epitope are linked to the mountain cedar pollen allergen Jun a 1. Jun a 1 is a glycoprotein highly homologous with Japanese cedar pollen glycoallergen, Cry j 1. Although it has been found that some plant complex type N-glycans are linked to Cry j 1, the occurrence of Lewis a epitope in the N-glycan moiety has not been proved yet. Hence, we reinvestigated the glycoform of the pollen allergen to find whether the Lewis a epitope(s) occur in the N-glycan moiety of Cry j 1. From the cedar pollen glycoallergen, the N-glycans were liberated by hydrazinolysis and the resulting sugar chains were N-acetylated and then coupled with 2-aminopyridine. Three pyridylaminated sugar chains were purified by reversed-phase HPLC and size-fractionation HPLC. The structures were analyzed by a combination of exo- and endo-glycosidase digestions, sugar chain mapping, and electrospray ionization mass spectrometry (ESI-MS). Structural analysis clearly indicated that Lewis a epitope (Gal beta 1-3(Fuc alpha 1-4)GlcNAc beta 1-), instead of the Gat beta 1-4(Fuc alpha 1-6)GlcNAc, occurs in the N-glycans of Cry j 1.

    DOI: 10.1271/bbb.69.1700

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  • Glycoform analysis of Japanese cedar pollen allergen, Cry j 1 国際誌

    M Maeda, M Kamamoto, K Hino, S Yamamoto, M Kimura, M Okano, Y Kimura

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   69 ( 9 )   1700 - 1705   2005年9月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    In our previous study (Y. Kimura et al., Biosci. Biotechnol, Biochem., 69, 137-144 (2005)), we found that plant complex type N-glycans harboring Lewis a epitope are linked to the mountain cedar pollen allergen Jun a 1. Jun a 1 is a glycoprotein highly homologous with Japanese cedar pollen glycoallergen, Cry j 1. Although it has been found that some plant complex type N-glycans are linked to Cry j 1, the occurrence of Lewis a epitope in the N-glycan moiety has not been proved yet. Hence, we reinvestigated the glycoform of the pollen allergen to find whether the Lewis a epitope(s) occur in the N-glycan moiety of Cry j 1. From the cedar pollen glycoallergen, the N-glycans were liberated by hydrazinolysis and the resulting sugar chains were N-acetylated and then coupled with 2-aminopyridine. Three pyridylaminated sugar chains were purified by reversed-phase HPLC and size-fractionation HPLC. The structures were analyzed by a combination of exo- and endo-glycosidase digestions, sugar chain mapping, and electrospray ionization mass spectrometry (ESI-MS). Structural analysis clearly indicated that Lewis a epitope (Gal beta 1-3(Fuc alpha 1-4)GlcNAc beta 1-), instead of the Gat beta 1-4(Fuc alpha 1-6)GlcNAc, occurs in the N-glycans of Cry j 1.

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  • Occurrence of lewis a epitope in N-glycans of a glycoallergen, Jun a 1, from mountain cedar (Juniperus ashei) pollen 国際誌

    Y Kimura, M Kamamoto, M Maeda, M Okano, M Yokoyama, K Kino

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   69 ( 1 )   137 - 144   2005年1月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    We have determined the structures of N-glycans linked to major allergens in the mountain cedar (Juniperus ashei) pollen, Jun a 1. First, two kinds of the pollen glycoallergen (Jun a I-A and Jun a 1-B) were purified from partially purified Jun a I by cation exchange chromatography. The N-glycans were liberated by hydrazinolysis from the two glycoallergens and the resulting sugar chains were N-acetylated and then coupled with 2-aminopyridine. Three pyridylaminated sugar chains were purified by reversed-phase HPLC and size-fractionation HPLC from Jun a 1-A and Jun a 1-B respectively. The structures were determined by a combination of exo- and endo-glycosidase digestions, two dimensional sugar chain mapping, and electrospray ionization mass spectrometry (ESI-MS) analysis. Structural analysis indicated that Lewis a epitope (Galbeta1-3(Fucalpha1-4)GlcNAcbeta1-) occurs in the N-glycans of the pollen allergens.

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  • Occurrence of lewis a epitope in N-glycans of a glycoallergen, Jun a 1, from mountain cedar (Juniperus ashei) pollen

    Y Kimura, M Kamamoto, M Maeda, M Okano, M Yokoyama, K Kino

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   69 ( 1 )   137 - 144   2005年1月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    We have determined the structures of N-glycans linked to major allergens in the mountain cedar (Juniperus ashei) pollen, Jun a 1. First, two kinds of the pollen glycoallergen (Jun a I-A and Jun a 1-B) were purified from partially purified Jun a I by cation exchange chromatography. The N-glycans were liberated by hydrazinolysis from the two glycoallergens and the resulting sugar chains were N-acetylated and then coupled with 2-aminopyridine. Three pyridylaminated sugar chains were purified by reversed-phase HPLC and size-fractionation HPLC from Jun a 1-A and Jun a 1-B respectively. The structures were determined by a combination of exo- and endo-glycosidase digestions, two dimensional sugar chain mapping, and electrospray ionization mass spectrometry (ESI-MS) analysis. Structural analysis indicated that Lewis a epitope (Galbeta1-3(Fucalpha1-4)GlcNAcbeta1-) occurs in the N-glycans of the pollen allergens.

    DOI: 10.1271/bbb.69.137

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  • N-glycan metabolism and plant cell differentiation and growth.

    Maeda Megumi, Kimura Y

    Trends in Glycoscience and Glycotechnology   17 ( 97 )   205 - 214   2005年

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  • Immunological activity of plant N-glycans and synthesis of neoglycopeptides and neoglycolipids carrying antigenic glycans

    M Sakaguchi, M Maeda, F Matsuda, M Kimura, M Okano, Y Kimura

    GLYCOBIOLOGY   14 ( 11 )   1120 - 1120   2004年11月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:OXFORD UNIV PRESS INC  

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  • N-glycans and endo-beta-N-acetylglucosaminidase in rice cultured cells

    M Maeda, Y Kimura

    GLYCOBIOLOGY   14 ( 11 )   1120 - 1120   2004年11月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:OXFORD UNIV PRESS INC  

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  • Roles of major oligosaccharides on Cry j 1 in human immunoglobulin E and T cell responses

    M. Okano, Y. Kimura, K. Kino, Y. Michigami, S. Sakamoto, Y. Sugata, M. Maeda, F. Matsuda, M. Kimura, T. Ogawa, K. Nishizaki

    Clinical and Experimental Allergy   34 ( 5 )   770 - 778   2004年5月

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    記述言語:英語  

    Background: We have demonstrated that carbohydrates in Cry j 1, the major allergen of Cryptomeria japonica pollen, play a major role in promoting Cry j 1-specific Th2 response. However, little is known as to whether the carbohydrates directly participate in allergic responses. Objective: We sought to determine whether Cry j 1-related oligosaccharides function as IgE and/or T cell epitopes. In addition, the regulatory effect of Cry j 1-related oligosaccharide on Cry j 1-specific T cell responses was investigated. Methods: Two monovalent oligosaccharides largely found on Cry j 1, Manα1- 6(Manα1-3) (Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc (M3FX), and GlcNAcβ1-2Manα1-6(GlcNAcβ1-2Manα1-3) (Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc (GN2M3FX) were prepared. Manα1-2Manα1-6(Manα1-2Manα1-3)Manα1- 6(Manα1-2Manα1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc (M9A) was used as control. Competitive inhibition ELISA for Cry j 1-specific IgE was performed using these oligosaccharides as inhibitors. In addition, T cell lines specific for Cry j 1 or purified protein derivative of Mycobacterium tubecurosis (PPD) were established, and cellular responses against these oligosaccharides were investigated in the presence or absence of the respective antigens. Results: Overall, neither M3FX nor GN2M3FX displayed inhibitory effect on the binding between IgE and Cry j 1. In addition, M3FX did not by itself stimulate Cry j 1 or PPD-specific T cells. However, M3FX significantly inhibited Cry j 1-induced proliferation and IL-4 production in Cry j 1-specific T cells. Such an inhibitory effect was not seen in PPD-specific T cell responses. Conclusion: These results suggest that Cry j 1-related oligosaccharides are not major epitopes for IgE or T cells. However, these oligosaccharides have a novel potential to inhibit Cry j 1-specific T cell responses selectively.

    DOI: 10.1111/j.1365-2222.2004.1948.x

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  • Roles of major oligosaccharides on Cry j 1 in human immunoglobulin E and T cell responses

    M. Okano, Y. Kimura, K. Kino, Y. Michigami, S. Sakamoto, Y. Sugata, M. Maeda, F. Matsuda, M. Kimura, T. Ogawa, K. Nishizaki

    Clinical and Experimental Allergy   34 ( 5 )   770 - 778   2004年5月

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    記述言語:英語  

    Background: We have demonstrated that carbohydrates in Cry j 1, the major allergen of Cryptomeria japonica pollen, play a major role in promoting Cry j 1-specific Th2 response. However, little is known as to whether the carbohydrates directly participate in allergic responses. Objective: We sought to determine whether Cry j 1-related oligosaccharides function as IgE and/or T cell epitopes. In addition, the regulatory effect of Cry j 1-related oligosaccharide on Cry j 1-specific T cell responses was investigated. Methods: Two monovalent oligosaccharides largely found on Cry j 1, Manα1- 6(Manα1-3) (Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc (M3FX), and GlcNAcβ1-2Manα1-6(GlcNAcβ1-2Manα1-3) (Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc (GN2M3FX) were prepared. Manα1-2Manα1-6(Manα1-2Manα1-3)Manα1- 6(Manα1-2Manα1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc (M9A) was used as control. Competitive inhibition ELISA for Cry j 1-specific IgE was performed using these oligosaccharides as inhibitors. In addition, T cell lines specific for Cry j 1 or purified protein derivative of Mycobacterium tubecurosis (PPD) were established, and cellular responses against these oligosaccharides were investigated in the presence or absence of the respective antigens. Results: Overall, neither M3FX nor GN2M3FX displayed inhibitory effect on the binding between IgE and Cry j 1. In addition, M3FX did not by itself stimulate Cry j 1 or PPD-specific T cells. However, M3FX significantly inhibited Cry j 1-induced proliferation and IL-4 production in Cry j 1-specific T cells. Such an inhibitory effect was not seen in PPD-specific T cell responses. Conclusion: These results suggest that Cry j 1-related oligosaccharides are not major epitopes for IgE or T cells. However, these oligosaccharides have a novel potential to inhibit Cry j 1-specific T cell responses selectively.

    DOI: 10.1111/j.1365-2222.2004.1948.x

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  • Role of pollen-derived oligosaccharides in human IgE and T cell responses in Japanese cedar pollinosis.

    Okano, M, Kimura, Y, Maeda, M, Michigamai, Kimura, M, Nishizaki. K

    Clin. Exp. Allergy   2004年

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  • Structural features of N-glycans linked to glycoproteins from oil palm pollen, an allergenic pollen

    Yoshinobu Kimura, Takeo Yoshiie, Woo Kwan Kit, Megumi Maeda, Mariko Kimura, Siang Hee Tan

    Bioscience, Biotechnology and Biochemistry   67 ( 10 )   2232 - 2239   2003年10月

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    記述言語:英語  

    The pollen of oil palm (Elaeis guineensis Jacq.) is a strong allergen and causes severe pollinosis in Malaysia and Singapore. In the previous study (Biosci. Biotechnol. Biochem., 64, 820-827 (2002)), from the oil palm pollens, we purified an antigenic glycoprotein (Ela g Bd 31 K), which is recognized by IgE from palm pollinosis patients. In this report, we describe the structural analysis of sugar chains linked to palm pollen glycoproteins to confirm the ubiquitous occurrence of antigenic N-glycans in the allergenic pollen. N-Glycans liberated from the pollen glycoprotein mixture by hydrazinolysis were labeled with 2-aminopyridine followed by purification with a combination of size-fractionation HPLC and reversed-phase HPLC. The structures of the PA-sugar chains were analyzed by a combination of two-dimensional sugar chain mapping, electrospray ionization mass spectrometry (ESI-MS), and tandem MS analysis, as well as exoglycosidase digestions. The antigenic N-glycan bearing α1-3 fucose and/or β1-2 xylose residues accounts for 36.9% of total N-glycans: GlcNAc2Man3Xyl1Fuc1GlcNAc 2 (24.6%), GlcNAc2Man3Xyl 1GlcNAc2 (4.4%), Man3Xyl1Fuc 1GlcNAc2 (1.1%), GlcNAc1Man3Xyl 1Fuc1GlcNAc2 (5.6%), and GlcNAc 1Man3Xyl1GlcNAc2 (1.2%). The remaining 63.1% of the total N-glycans belong to the high-mannose type structure: Man9GlcNAc2 (5.8%), Man8GlcNAc2 (32.1%), Man7GlcNAc2 (19.9%), Man6GlcNAc2 (5.3%).

    DOI: 10.1271/bbb.67.2232

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  • Structural features of N-glycans linked to glycoproteins from oil palm pollen, an allergenic pollen 国際誌

    Yoshinobu Kimura, Takeo Yoshiie, Woo Kwan Kit, Megumi Maeda, Mariko Kimura, Siang Hee Tan

    Bioscience, Biotechnology and Biochemistry   67 ( 10 )   2232 - 2239   2003年10月

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    記述言語:英語  

    The pollen of oil palm (Elaeis guineensis Jacq.) is a strong allergen and causes severe pollinosis in Malaysia and Singapore. In the previous study (Biosci. Biotechnol. Biochem., 64, 820-827 (2002)), from the oil palm pollens, we purified an antigenic glycoprotein (Ela g Bd 31 K), which is recognized by IgE from palm pollinosis patients. In this report, we describe the structural analysis of sugar chains linked to palm pollen glycoproteins to confirm the ubiquitous occurrence of antigenic N-glycans in the allergenic pollen. N-Glycans liberated from the pollen glycoprotein mixture by hydrazinolysis were labeled with 2-aminopyridine followed by purification with a combination of size-fractionation HPLC and reversed-phase HPLC. The structures of the PA-sugar chains were analyzed by a combination of two-dimensional sugar chain mapping, electrospray ionization mass spectrometry (ESI-MS), and tandem MS analysis, as well as exoglycosidase digestions. The antigenic N-glycan bearing α1-3 fucose and/or β1-2 xylose residues accounts for 36.9% of total N-glycans: GlcNAc2Man3Xyl1Fuc1GlcNAc 2 (24.6%), GlcNAc2Man3Xyl 1GlcNAc2 (4.4%), Man3Xyl1Fuc 1GlcNAc2 (1.1%), GlcNAc1Man3Xyl 1Fuc1GlcNAc2 (5.6%), and GlcNAc 1Man3Xyl1GlcNAc2 (1.2%). The remaining 63.1% of the total N-glycans belong to the high-mannose type structure: Man9GlcNAc2 (5.8%), Man8GlcNAc2 (32.1%), Man7GlcNAc2 (19.9%), Man6GlcNAc2 (5.3%).

    DOI: 10.1271/bbb.67.2232

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  • Purification and characterization of 31-kDa palm pollen glycoprotein (Ela g Bd 31 K), which is recognized by IgE from palm pollinosis patients 国際誌

    Yoshinobu Kimura, Megumi Maeda, Mariko Kimura, Oi Ming Lai, Siang Hee Tan, Sook Mei Hon, Fook Tim Chew

    Bioscience, Biotechnology and Biochemistry   66 ( 4 )   820 - 827   2002年

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    記述言語:英語  

    A basic glycoprotein, which was recognized by IgE from oil palm pollinosis patients, has been purified from oil palm pollen (Elaeis guineensis Jacq.), which is a strong allergen and causes severe pollinosis in Malaysia and Singapore. Soluble proteins were extracted from defatted palm pollen with both Tris-HCl buffer (pH 7.8) and Na-acetate buffer (pH 4.0). The allergenic glycoprotein was purified from the total extract to homogeneity with 0.4% yield by a combination of DEAE- and CM-cellulose, SP-HPLC, and gel filtration. The purified oil palm pollen glycoprotein with molecular mass of 31 kDa was recognized by the β1-2 xylose specific antibody, suggesting this basic glycoprotein bears plant complex type N-glycan(s). The palm pollen basic glycoprotein, designated Ela g Bd 31 K, was recognized by IgE of palm pollinosis patients, suggesting Ela g Bd 31 K should be one of the palm pollen allergens. The preliminary structural analysis of N-glycans linked to glycoproteins of palm pollens showed that the antigenic N-glycans having α1-3 fucose and β1-2 xylose residues (GlcNAc2∼0Man3Xyl1Fuc1∼0GlcNAc2) actually occur on the palm pollen glycoproteins, in addition to the high-mannose type structures (Man9∼5GlcNAc2). © 2002 by Japan Society for Bioscience, Biotechnology, and Agrochemistry.

    DOI: 10.1271/bbb.66.820

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  • Purification and characterization of 31-kDa palm pollen glycoprotein (Ela g Bd 31 K), which is recognized by IgE from palm pollinosis patients

    Yoshinobu Kimura, Megumi Maeda, Mariko Kimura, Oi Ming Lai, Siang Hee Tan, Sook Mei Hon, Fook Tim Chew

    Bioscience, Biotechnology and Biochemistry   66 ( 4 )   820 - 827   2002年

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    記述言語:英語  

    A basic glycoprotein, which was recognized by IgE from oil palm pollinosis patients, has been purified from oil palm pollen (Elaeis guineensis Jacq.), which is a strong allergen and causes severe pollinosis in Malaysia and Singapore. Soluble proteins were extracted from defatted palm pollen with both Tris-HCl buffer (pH 7.8) and Na-acetate buffer (pH 4.0). The allergenic glycoprotein was purified from the total extract to homogeneity with 0.4% yield by a combination of DEAE- and CM-cellulose, SP-HPLC, and gel filtration. The purified oil palm pollen glycoprotein with molecular mass of 31 kDa was recognized by the β1-2 xylose specific antibody, suggesting this basic glycoprotein bears plant complex type N-glycan(s). The palm pollen basic glycoprotein, designated Ela g Bd 31 K, was recognized by IgE of palm pollinosis patients, suggesting Ela g Bd 31 K should be one of the palm pollen allergens. The preliminary structural analysis of N-glycans linked to glycoproteins of palm pollens showed that the antigenic N-glycans having α1-3 fucose and β1-2 xylose residues (GlcNAc2∼0Man3Xyl1Fuc1∼0GlcNAc2) actually occur on the palm pollen glycoproteins, in addition to the high-mannose type structures (Man9∼5GlcNAc2). © 2002 by Japan Society for Bioscience, Biotechnology, and Agrochemistry.

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▼全件表示

講演・口頭発表等

  • 酸性及び細胞質ペプチド: N-グリカナーゼ二重欠損 A. thaliana遊離N-グリカン構造解析

    奥村陸, 白井佐保子, 三﨑亮, 梶浦裕之, 藤山和仁, 木村吉伸, 前田恵

    学会創立100周年記念 中四国支部・西日本支部合同大会(第66回 講演会)(高知) 

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    開催年月日: 2023年9月21日 - 2023年9月22日

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    板野紗月, 沓野那緒, 藤原美智子, 加来田博貴, 木村吉伸, 前田恵

    第42回日本糖質学会年会(鳥取) 

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    開催年月日: 2023年9月7日 - 2023年9月9日

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    井口 夢香, 堀口 星, 前田 恵, 木村 吉伸

    第95回日本生化学会大会(名古屋) 

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    中野海藍, 小坂将太, 前田恵, 木村吉伸

    第95回日本生化学会大会(名古屋) 

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    第95回日本生化学会大会(名古屋) 

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    奥村 陸, 白井 佐保子, 前田 恵, 三﨑 亮, 梶浦裕之, 藤山 和仁, 木村 吉伸

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    児玉 怜央, 岡本 尚子, 山本 千晴, 前田 恵, 木村 吉伸

    第41回日本糖質学会年会(大阪) 

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    開催年月日: 2022年9月29日 - 2022年10月1日

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    Inetrnational Mesothelioma Interest Group. Congress 2008.  2008年 

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    Inetrnational Mesothelioma Interest Group. Congress 2008.  2008年 

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    Inetrnational Mesothelioma Interest Group. Congress 2008.  2008年 

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    International Investigative Dermatology 2008  2008年 

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    Inetrnational Mesothelioma Interest Group. Congress 2008.  2008年 

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    Inetrnational Mesothelioma Interest Group. Congress 2008.  2008年 

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    Inetrnational Mesothelioma Interest Group. Congress 2008.  2008年 

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    第15回日本免疫毒性学会学術大会  2008年 

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    第15回日本免疫毒性学会学術大会  2008年 

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    第77回日本衛生学会総会  2007年 

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    18th Japan-China-Korea Joint Conference on Occupational Health  2007年 

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    18th Japan-China-Korea Joint Conference on Occupational Health  2007年 

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    第80回日本生化学会大会  2007年 

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    Professor E. Carwile LeRoy Memorial International Workshop on Scleroderma  2007年 

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    66th Annual Meetingof the Japanese Cancer Association  2007年 

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    第48回日本肺癌学会総会  2007年 

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    第37回日本免疫学会総会  2007年 

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  • SScへの環境因子としての珪肺症例における末梢でのCD4+25+ 分画の機能減少 Reduced function of the peripheral CD4+25+ fraction in silicosis patients exposed to environmental factors for SSc.

    第37回日本免疫学会総会  2007年 

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    第3回中国皮膚科研究セミナー  2007年 

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    第37回日本免疫学会総会  2007年 

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    第7回分子予防環境医学研究会  2007年 

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    第55回日本職業・災害医学会学術大会  2007年 

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    第14回石綿・中皮腫研究会  2007年 

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  • 珪肺症例におけるCD4+25+分画における慢性活性化T細胞、Fas分子発現の検討

    第35回日本臨床免疫学会総会  2007年 

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  • アスベスト曝露症例末梢血CD3+T細胞におけるT細胞受容体Vβ発現の検討と実験系における検証

    第48回日本肺癌学会総会  2007年 

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  • 悪性中皮腫症例末梢血CD4+T細胞におけるBcl-2高発現と実験的検証

    第48回日本肺癌学会総会  2007年 

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  • Immunological effects of asbestos.

    66th Annual Meetingof the Japanese Cancer Association  2007年 

     詳細を見る

  • The functional impairment of NK cells due to the long-term exposure to asbestos.

    18th Japan-China-Korea Joint Conference on Occupational Health  2007年 

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  • Immunological Effects of Asbestos.

    18th Japan-China-Korea Joint Conference on Occupational Health  2007年 

     詳細を見る

  • Reduced function of the peripheral CD4+25+ fraction in silicosis patients exposed to environmental factors for SSc.

    Professor E. Carwile LeRoy Memorial International Workshop on Scleroderma  2007年 

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  • 珪肺症例におけるCD4+25+分画における慢性活性化T細胞、Fas分子発現の検討

    第14回日本免疫毒性学会学術大会  2007年 

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  • アスベスト曝露ヒトポリクローナルT細胞株における炎症性サイトカインIL-10およびTGF-β1の産生誘導

    第14回日本免疫毒性学会学術大会  2007年 

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  • 珪肺症例末梢血CD4+25+分画におけるFas分子発現

    第38回日本職業・環境アレルギー学会・学術大会  2007年 

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  • スギ花粉アレルゲンCry j1およびJun a1に結合する糖タンパク質糖鎖の構造特性と免疫活性

    第38回日本職業・環境アレルギー学会・学術大会  2007年 

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  • 植物糖タンパク質糖鎖の代謝機構とアレルゲン糖鎖の構造・機能解析

    第18回日本農芸化学会中四国支部講演会  2007年 

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  • マウスパラコート肺モデルでの肺障害初期に関与する遺伝子の検討.

    第29回日本フリーラジカル学会/日本過酸化脂質・フリーラジカル学会第31回大会(合同学会)  2007年 

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  • ヒトポリクローナルリンパ球MT-2株のアスベスト曝露による遺伝子発現変化

    第80回日本産業衛生学会  2007年 

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  • アスベスト長期曝露下ヒトNK細胞株における2B4(CD244)依存性細胞障害性および脱顆粒の低下

    第80回日本産業衛生学会  2007年 

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  • マイナス電荷空気環境の生体影響の観察

    第16回日本臨床環境医学会  2007年 

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  • マウスパラコート肺モデル早期での遺伝子発現とサイトカインレベルの変化

    第34回日本トキシコロジー学会学術年会  2007年 

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    会議種別:ポスター発表  

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  • 珪酸曝露:ヒト免疫系に及ぼす影響

    第18回日本微量元素学会学術集会  2007年 

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  • 石綿長期曝露後のヒトNK細胞株YT-A1におけるNK活性化レセプター2B4の機能低下

    第77回日本衛生学会総会  2007年 

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  • アスベスト長期曝露によるNK細胞の機能変化

    第77回日本衛生学会総会  2007年 

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  • 石綿起因疾患症例リンパ球における病態関連遺伝子発現の解析

    第13回日本免疫毒性学会学術大会  2006年 

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  • Alterations of Fas and Fas-related molecules in patients with silicosis.

    The 28th International Congress o Occupational Health  2006年 

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  • Studies on the deglycosylation mechanism in plant cells.

    20th IUBMB Internationla Cngress of Biochemistry and Molecular Biology and 11th PAOBMB Congress  2006年 

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  • 石綿曝露下長期培養後のヒトNK細胞株YT-A1における機能変化

    第6回分子予防環境医学研究会  2006年 

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  • アスベスト長期曝露によるNK細胞の細胞傷害性低下およびNKレセプターの発現異常

    第36回日本免疫学会総会学術集会  2006年 

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  • Alterations of Fas and Fas-related molecules in patients with silicosis.

    The 28th International Congress o Occupational Health  2006年 

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  • Studies on the deglycosylation mechanism in plant cells.

    20th IUBMB Internationla Cngress of Biochemistry and Molecular Biology and 11th PAOBMB Congress  2006年 

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  • 珪肺症例における末梢血CD4+25+分画の機能解析

    第54回日本職業・災害医学会  2006年 

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  • ヒトHTLV-1不死化多クローン性T細胞株MT-2のアスベスト誘導アポトーシス抵抗性亜株の性質の検討

    第6回分子予防環境医学研究会  2006年 

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  • HTLV-1不死化ヒト多クローン性T細胞株/MT-2における石綿誘導細胞死抵抗性と制御性T細胞機能について

    第68回日本血液学会/第48回日本臨床血液学会合同総会  2006年 

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  • ヒトHTLV-1不死化多クローン性T細胞株MT-2のアスベスト誘導アポトーシス抵抗性亜株の性状

    第13回石綿・中皮腫研究会  2006年 

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  • 珪肺症例における制御性T細胞分画の機能解析

    第15回日本臨床環境医学会  2006年 

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  • アスベスト関連発癌と免疫影響.フォーカス講演「アスベストと悪性中皮腫」―発症の基礎と臨床の実際―

    第17回日本微量元素学会  2006年 

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  • 環境免疫学~話題の病態の背景に迫る-アスベスト(珪酸塩)と珪酸の免疫影響の比較から-

    第37回日本職業・環境アレルギー学会総会・学術大会  2006年 

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  • アスベスト長期曝露によるNK細胞の機能変化

    第13回日本免疫毒性学会学術大会  2006年 

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  • 珪肺症例における制御性T細胞分画の機能解析

    第13回日本免疫毒性学会学術大会  2006年 

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▼全件表示

受賞

  • 研究集会助成事業

    2022年12月   公益財団法人八雲環境科学振興財団  

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  • 両備檉園記念財団 生物学研究奨励賞

    2021年10月  

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  • 49th Society of Toxicology (USA); Immunotoxicology Specialty Section, HESI Immunotoxicology, Young Investigator Travel Award

    2010年3月  

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  • 2006年度日本農芸化学会大会B.B.B.論文賞 Glycoform analysis of Japanese cedar pollen allergen, Cry j1 Biosci. Biotechnol. Biochem., 69, 1700-1705 (2005)

    2007年  

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    受賞国:日本国

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  • 第18回日本農芸化学会中四国支部講演会 2007年度農芸化学会中四国支部奨励賞 植物糖タンパク質糖鎖の代謝機構とアレルゲン糖鎖の構造・機能解析.

    2007年  

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    受賞国:日本国

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共同研究・競争的資金等の研究

  • 新規食品素材「アクアファバ」に含まれる遊離型オリゴ糖・ペプチドの構造と機能解析

    研究課題/領域番号:21K02101  2021年04月 - 2024年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    木村 万里子, 前田 恵, 山下 弘高, 吉田 和利, 藤田 裕之

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    配分額:4160000円 ( 直接経費:3200000円 、 間接経費:960000円 )

    ①豆煮汁からの遊離型オリゴ糖の精製および化学構造解析:6種の豆類(大豆,ひよこまめ, 手亡, 小豆, えんどう, レンズまめ)から,一般的な餡の製造工程に準じて2時間加熱して煮汁を調製し,それらの凍結乾燥物を得た。次に,各凍結乾燥物から,50%エタノール分画,陽イオン交換によりオリゴ糖画分を調製し,順相HPLC-RI及びLC-ESI-MSでラフィノース族オリゴ糖の定量分析を行った。その結果,ひよこまめ煮汁はラフィノースの,大豆,ひよこまめ,小豆およびえんどうの煮汁はスタキオースの良い供給源として利用できる可能性が認められた。小豆と手亡の煮汁中にN-グリカン(M8,M3FX)の存在も認められたものの,今回定量には至らなかった。
    ②豆煮汁由来オリゴ糖の機能性解析:豆煮汁オリゴ糖画分のプレバイオティック効果は,4種のビフィズス菌において有意に認められたものの(p<0.01),N-グリカンの増殖活性は認められなかった。
    ③ 豆煮汁からのACE阻害ペプチドの精製および化学構造・機能解析:6種の豆煮汁から,上述①と同じ手法でペプチド画分を調製し,さらにODSカラムと逆相HPLCに供してACE阻害活性ペプチドを単離した。活性ペプチドの構造解析は,MS/MS分析,プロテインシークエンサーにより行った。ACE阻害活性測定は,合成基質から遊離する馬尿酸をHPLCで定量する手法により行った。
    ④ 各種「アクアファバ」の泡の物性および嗜好性:3種(ひよこ豆、小豆、手亡)の豆煮汁を一定条件下で泡立て光学顕微鏡観察を行ったところ,豆の種類により泡沫の状態(安定性や泡直径など)が異なることが認められた。

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  • 機能性糖鎖ポリマーの合成と花粉アレルゲン特異的Th2免疫応答の抑制活性解析

    研究課題/領域番号:18K05559  2018年04月 - 2021年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    前田 恵

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    配分額:4420000円 ( 直接経費:3400000円 、 間接経費:1020000円 )

    植物抗原性糖鎖の骨格構造であるMan3Fuc1Xyl1GlcNAc2(M3FX)はスギ花粉症患者のTh2免疫応答を有意に抑制することが明らかになっているが,その抑制メカニズムは未だ解明されていない。また,スギやヒノキ花粉アレルゲンに特徴的に発現しているM3FXの非還元末端がルイスa抗原で修飾された糖鎖のTh2免疫応答における活性については明らかにされていない。そこで昨年度は,オオカナダモの糖タンパク質からルイスa抗原含有糖鎖を多量精製し,多価に結合させた糖鎖ポリマーを作成し,ヒト単球細胞株THP-1細胞から誘導した樹状細胞様細胞株について抗原提示やmaturationへの糖鎖ポリマーによる影響を解析してきた。しかしながら,オオカナダモはエンドトキシン混入の可能性があり,これまではルイスa抗原含有糖鎖の多量調製源としてオオカナダモを用いていたが,免疫活性解析に適さない恐れがあった。そこで本研究では貯蔵タンパク質を多く含み,ヒトが食用としているイモ類に着目し,4種類のイモを用いて貯蔵タンパク質の糖鎖構造解析を行った。まず,サトイモ(親芋および子芋),サツマイモ,ナガイモの貯蔵タンパク質から,ペプシン消化,陽イオン交換,ゲルろ過により糖ペプチドを調製した。ヒドラジン分解,ピリジルアミノ化(PA化),RP-/SF-HPLCによりPA糖鎖を精製した後,エンドおよびエキソグリコシダーゼを用いた逐次酵素消化法,MS, MS/MSにより糖鎖構造解析を行った。その結果,全てのイモ類貯蔵タンパク質糖鎖の主要構造は植物抗原性糖鎖であった。また,サトイモ貯蔵タンパク質にはルイスa抗原含有糖鎖がオオカナダモ由来糖タンパク質と同程度(約0.4 nmol/g)結合していた。

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  • 抗花粉症薬剤の開発を目指した機能性糖鎖ポリマーの作製と細胞性免疫活性解析

    研究課題/領域番号:15K07841  2015年04月 - 2018年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    前田 恵, 木村 吉伸, 岡野 光博

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    配分額:4940000円 ( 直接経費:3800000円 、 間接経費:1140000円 )

    植物複合型糖鎖によるスギ花粉アレルゲンCry j1特異的Th2免疫応答の抑制機構を解明するため,3種類の糖鎖ポリマー(植物複合型,ハイマンノース型,動物複合型)を合成し,樹状細胞の分化抑制を解析した。新規ヒノキ花粉アレルゲンCha o3の糖鎖構造解析を行い,その構造的特徴は日本スギ花粉アレルゲンCry j1及びマウンテンシダーアレルゲンJun a1と非常に高い相同性を有することを明らかにした。

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  • 抗花粉症薬剤の開発を目指した機能性糖鎖ポリマーの作製と細胞性免疫活性解析(国際共同研究強化)

    研究課題/領域番号:15KK0282  2015年 - 2017年

    日本学術振興会  科学研究費助成事業 国際共同研究加速基金(国際共同研究強化)  国際共同研究加速基金(国際共同研究強化)

    前田 恵

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    配分額:14040000円 ( 直接経費:10800000円 、 間接経費:3240000円 )

    植物細胞に遍在する遊離N-グリカン(FNGs)は,糖タンパク質や糖ペプチドからペプチド:N-グリカナーゼ(PNGase)やエンド-β-N-アセチルグルコサミニダーゼ(ENGase)により生成され植物の分化・生長や果実熟成を促進するホルモン様活性を有すると推定されている。また近年,植物細胞には様々な機能未知のレクチンが発現しており,それらのいくつかは環境ストレス(塩,乾燥,気温,微生物感染など)により発現誘導することが報告されている。そこで本研究では,(1)糖鎖遊離酵素欠損A.thalianaのストレス(塩や病原体)感受性の解析と(2)糖鎖ポリマーによるレクチンのスクリーニングを試みた。

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  • 植物抗原性糖鎖ポリマーによる自然免疫を介したTh2分化抑制の解析

    研究課題/領域番号:24580494  2012年04月 - 2015年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    前田 恵, 木村 吉伸, 大槻 剛巳

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    配分額:5460000円 ( 直接経費:4200000円 、 間接経費:1260000円 )

    スギ花粉アレルゲンによるTh2応答を抑制する植物糖タンパク質の植物抗原性糖鎖は,花粉症治療薬としての利用が期待されており,本研究では免疫活性を高めた糖鎖ポリマー作製を目的として,植物抗原性糖鎖を銀杏種子より多量精製する親水性クロマト法を確立した。次いでルイスa抗原を含有している植物抗原性糖鎖を水草から見出した。更には植物抗原性糖鎖を多価に結合した糖鎖ポリマーを作製し,樹状細胞の分化抑制を解析した。

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  • 雑豆由来オリゴ糖鎖の機能性開発と食品への応用

    研究課題/領域番号:23500944  2011年04月 - 2016年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    木村 万里子, 長澤 治子, 前田 恵

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    配分額:5200000円 ( 直接経費:4000000円 、 間接経費:1200000円 )

    本研究は、雑豆(大豆と落花生を除く豆類)の機能性食材としての価値を検討することを目的としている。まず、小豆と手亡(白いんげん豆)から均一性の高い多量の糖ペプチド・オリゴ糖を調製する方法を確立した。次に、調製したオリゴ糖をγ-ポリグルタミン酸にカップリングさせたネオ糖ペプチドを作り、それらの抗原提示細胞の活性化、T細胞分化・生育、サイトカイン産生に及ぼす影響を調べたところ、ヒト単球を活性化することが認められた。さらに、小豆や手亡に含まれる遊離型オリゴ糖の糖組成分析を行うとともに、プレバイオティクス活性を調べた。以上の結果から、小豆や手亡には高い機能性を有するオリゴ糖が存在することが示唆された。

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  • T細胞CXCR3発現の悪性中皮腫早期診断・予防マーカーとしての有用性

    研究課題/領域番号:22700933  2010年 - 2011年

    日本学術振興会  科学研究費助成事業 若手研究(B)  若手研究(B)

    前田 恵

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    配分額:4030000円 ( 直接経費:3100000円 、 間接経費:930000円 )

    アスベスト起因性胸膜プラークおよび悪性中皮腫症例の末梢CD4+ T細胞はケモカインレセプターCXCR3発現が健常人に比べて有意に低下しており, IFN-γ発現は悪性中皮腫症例のみで有意に低下していることを見出した.実験的にヒトCD4+ T細胞へのアスベスト慢性曝露はCXCR3とIFN-γ発現誘導を阻害する一方で, IL-17発現を高めることを示し,免疫担当細胞へのアスベスト曝露は抗腫瘍免疫機能の減弱に加えて,腫瘍の増殖促進に関与する可能性を示唆した.

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  • 室内気質の免疫影響に関する実験システムの構築

    研究課題/領域番号:21659161  2009年 - 2011年

    日本学術振興会  科学研究費助成事業 挑戦的萌芽研究  挑戦的萌芽研究

    大槻 剛巳, 西村 泰光, 武井 直子, 松崎 秀紀, 李 順姫, 前田 恵

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    配分額:3270000円 ( 直接経費:3000000円 、 間接経費:270000円 )

    マイナス荷電粒子優位の室内空気室を構築して,健常人ボランティアにより2.5時間滞在,ならびに夜間のみ2週間滞在型試験を行ってきた結果,短期間のインターロイキン2の軽微な,しかし有意な上昇と,中期間(2週間)でのナチュラルキラー細胞の活性増強を観察していたので,今回,実験的にそのシステムの構築と,検証を行った結果,マイナス荷電粒子の優位の程度は,オーダーは異なっていたが,実験的にこの環境がヘルパーT細胞の活性化とナチュラルキラー細胞の活性化を誘導することが判明した。

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  • アスベスト発癌に影響する免疫動態の解明

    研究課題/領域番号:20390178  2008年 - 2010年

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    大槻 剛巳, 西村 泰光, 前田 恵, 熊谷 直子, 村上 周子

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    配分額:18070000円 ( 直接経費:13900000円 、 間接経費:4170000円 )

    2005年のクボタショック以来アスベスト曝露による悪性腫瘍の発生が問題となっているが,アスベスト曝露の免疫影響が腫瘍発生の素地を形成する可能性を考え検討してきた。その結果,実験系での免疫担当細胞(細胞株や健常人からの新鮮採取細胞)へのアスベスト曝露によって腫瘍免疫の減衰を示唆する所見が得られてきた。今後はその動態を改善することによって予防的分子標的を同定するなどに応用したい。

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  • 環境中物質の免疫影響スクリーニングシステム構築

    研究課題/領域番号:19659153  2007年 - 2008年

    日本学術振興会  科学研究費助成事業 萌芽研究  萌芽研究

    大槻 剛巳, 西村 泰光, 前田 恵, 村上 周子, 熊谷 直子

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    配分額:3300000円 ( 直接経費:3300000円 )

    本研究「環境中物質の免疫影響スクリーニングシステム構築」では,以下を考案した。環境中物質の免疫影響のスクリーニングシステムとしては,最終的には,インフォームドコンセントを得,倫理委員会の承認を受けた上で,曝露を受けた可能性のある症例等の末梢血を一定量採取し,可能性のある物質のin vitro曝露系の構築が必要である。この場合,アレルギーや感作という側面では,Th1/2バランスのシフトが重要になるであろう。また,自己寛容の破綻という側面では,我々が検討してきたFas分子発現機構や機能に関する解析システムの簡易化も必要であろうし,CD4+25+FoxP3+制御性T細胞の機能解析も重要であろう。加えて,腫瘍発生という観点からも,その免疫毒性〜免疫系への影響を検証するのは,分子予防標的な探索にも必要であろうと考えられる。特に,腫瘍発生についても,腫瘍を認識するNK細胞,cytotoxic T細胞,そしてそれを制御するCD4+25+FoxP3+制御性T細胞の機能解析は重要であろうと考えられる。これらの中で,現在は,珪酸曝露における制御性T細胞とFas分子の関連,そしてそこから生じる自己寛容の異常について,そしてアスベスト曝露による制御性T,NK,NKT,CD8+T細胞などの変化を捉え,変化に関与する分子の同定を行ってきた。これらの検討にて,シリカによる自己寛容の破綻にはFAS分子を介した制御性T細胞の活性化とその早期細胞死に加えて反応性T細胞の活性化による制御性T細胞分画への混入が重要と考えられ,またこれらの病態を反映する因子として可溶性IL-2受容体は重要であること,反対に可溶性CD40ligandはそれほど重要ではないことなどが判明してきた。またアスベストではIL-10やTGFβが重要であろうと判明してきた。

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  • アスベスト曝露が引き起こす腫瘍免疫の減衰と悪性中皮腫の発生

    研究課題/領域番号:19790411  2007年 - 2008年

    日本学術振興会  科学研究費補助金  若手研究(B)

    前田 恵

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    資金種別:競争的資金

    HTLV-1不死化T細胞株MT-2にアスベスト繊維の一種であるクリソタイルを低濃度長期曝露しクリソタイル耐性を獲得した細胞株を樹立することにより,アスベスト繊維曝露はCD4陽性T細胞のIFN-γ産生を抑制する一方で,IL-10,TGF-β1産生を促進することを示唆した。耐性株ではTh1型のT細胞に発現しており細胞遊走に関わるケモカインレセプターCXCR3の発現低下も生じており,アスベスト曝露による抗腫瘍免疫の減衰を推察できた。

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  • 石綿曝露者における発癌予防に関わる免疫学的分子標的の解析

    研究課題/領域番号:18390186  2006年 - 2007年

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    大槻 剛巳, 西村 泰光, 前田 恵, 村上 周子

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    配分額:16830000円 ( 直接経費:14700000円 、 間接経費:2130000円 )

    本研究「石綿曝露者における発癌予防に関わる免疫学的分子標的の解析」では,アスベストの免疫担当細胞への影響を観察することから始まり,そこで得られた所見を,アスベスト曝露者における曝露指標あるいは,悪性中皮腫発生などに伴う,担癌指標として活用できないかどうか,加えて,それら免疫担当細胞の変化が,腫瘍発生に関連するとして,その免疫学的変化を生じなくすることが,発癌予防につながらないかどうかを検討することを目的とした。その結果,1)アスベスト曝露が制御性T細胞機能を充進させる結果,腫瘍免疫の減衰を齎す可能性があること。2)ケモカイン受容体のひとつであるCXCR3の膜発現低下が,細胞株モデルのアスベスト低濃度長期曝露モデル,あるいは,freshly isolated peripheral T cell derived from health donorsを用いたin vitro実験系でも確認され,これはアスベスト曝露による胸膜プラークを示す症例,さらに強く悪性中皮腫症例で,同様の減少が認められること。3)TGFβやIL-10は,アスベスト長期曝露T細胞株でも,また,悪性中皮腫症例でも高値を示し,担癌指標となる可能性と,上記の制御性T細胞機能にも関連することが示唆されること。4)TGFβはまた,それのみでも,細胞増殖系のシグナルを制御抑制し,そのことが制御性T細胞などの機能充進につながる可能性があること。5)アスベスト長期曝露T細胞株モデルでは,NFkBの発現が低下しており,これもまた増殖能よりも分化能ひいては機能充進へと関連する可能性があること,等が判明した。今後は,アスベスト曝露者の早期診断指標かつ分子予防標的として制御性T細胞関連の分子の役割について,研究を進めていく予定である。

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  • 植物糖タンパク質糖鎖の構造・機能解析 および植物エンドグリコシダーゼの機能特性に関する研究

    2003年 - 2006年

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    資金種別:競争的資金

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