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

PubMed

researchmap

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

PubMed

researchmap

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

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

PubMed

researchmap

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

PubMed

researchmap

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

PubMed

researchmap

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

PubMed

researchmap

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

PubMed

researchmap

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

PubMed

researchmap

記述言語：日本語   出版者・発行元：(公社)日本産業衛生学会  

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3892/or.2018.6481

PubMed

researchmap

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.

DOI： 10.1093/jb/mvy029

PubMed

researchmap

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

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3390/ijms19020504

PubMed

researchmap

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

Scopus

PubMed

researchmap

DOI： 10.11482/KMJ-E44(1)33

researchmap

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

Web of Science

PubMed

researchmap

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

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3892/ijo.2017.3991

PubMed

researchmap

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.

DOI： 10.1093/jb/mvw089

Scopus

PubMed

researchmap

記述言語：日本語   出版者・発行元：(公社)日本産業衛生学会  

researchmap

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

Scopus

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3892/ijo.2016.3776

PubMed

researchmap

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.

DOI： 10.1080/09168451.2017.1320519

Web of Science

PubMed

researchmap

記述言語：日本語  

researchmap

記述言語：日本語  

researchmap

記述言語：日本語   出版者・発行元：日本繊維状物質研究協会  

CiNii Article

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3109/1547691X.2016.1143539

PubMed

researchmap

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

Scopus

PubMed

researchmap

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.

DOI： 10.3892/ol.2016.4412

PubMed

researchmap

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).

DOI： 10.1080/09168451.2015.1079479

Web of Science

PubMed

researchmap

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

Scopus

PubMed

researchmap

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

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.imbio.2015.07.006

PubMed

researchmap

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

Scopus

researchmap

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

Scopus

researchmap

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.

DOI： 10.1080/09168451.2015.1034653

Web of Science

PubMed

researchmap

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。

DOI： 10.1007/s10719-015-9596-4

researchmap

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.

DOI： 10.1111/gtc.12246

Scopus

PubMed

researchmap

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

Scopus

PubMed

researchmap

記述言語：日本語  

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1371/journal.pone.0132373

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1155/2015/238431

PubMed

researchmap

DOI： 10.1007/s10719-015-9596-4

researchmap

DOI： 10.1007/s10719-015-9596-4

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3892/ijo.2014.2682

PubMed

researchmap

記述言語：日本語  

researchmap

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

PubMed

researchmap

記述言語：日本語   出版者・発行元：科学評論社  

CiNii Article

CiNii Books

researchmap

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

記述言語：日本語  

researchmap

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

Web of Science

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1155/2014/670140

PubMed

researchmap

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

CiNii Article

researchmap

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

PubMed

researchmap

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.

DOI： 10.1271/bbb.130303

Web of Science

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1165/rcmb.2012-0134OC

PubMed

researchmap

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.

DOI： 10.1007/s10719-012-9441-y

Web of Science

PubMed

researchmap

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

PubMed

researchmap

掲載種別：研究論文（学術雑誌）  

researchmap

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。

DOI： 10.14869/toxpt.40.1.0.1114.0

CiNii Article

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1271/bbb.120463

PubMed

researchmap

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

PubMed

researchmap

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

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1155/2012/492608

PubMed

researchmap

researchmap

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.

DOI： 10.1165/rcmb.2010-0435OC

Scopus

PubMed

researchmap

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

CiNii Article

CiNii Books

researchmap

その他リンク： http://igakkai.kms-igakkai.com/wp/wp-content/uploads/2011ky/ippannkyouyou37.47-59.2011.pdf

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

Scopus

PubMed

researchmap

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.

Scopus

researchmap

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

PubMed

researchmap

記述言語：日本語  

researchmap

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.

DOI： 10.1271/bbb.110148

Web of Science

PubMed

researchmap

記述言語：日本語  

researchmap

記述言語：英語  

researchmap

記述言語：英語  

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1155/2011/481439

PubMed

researchmap

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。

DOI： 10.1007/s10719-011-9334-5

researchmap

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.

Scopus

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

researchmap

DOI： 10.3109/1547691X.2010.512579

PubMed

researchmap

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

PubMed

researchmap

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。

DOI： 10.1093/jb/mvq102

PubMed

researchmap

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)

CiNii Article

CiNii Books

researchmap

その他リンク： http://search.jamas.or.jp/link/ui/2010218223

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

Scopus

PubMed

researchmap

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

CiNii Article

CiNii Books

researchmap

その他リンク： http://id.nii.ac.jp/1162/00001517/

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

CiNii Article

CiNii Books

researchmap

その他リンク： http://id.nii.ac.jp/1162/00001524/

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.

DOI： 10.1007/s12199-008-0063-8

Scopus

PubMed

researchmap

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)

researchmap

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

Scopus

researchmap

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

Scopus

PubMed

researchmap

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

Scopus

researchmap

記述言語：日本語   出版者・発行元：山口大学医学会  

CiNii Article

CiNii Books

researchmap

記述言語：日本語   出版者・発行元：(一社)日本衛生学会  

CiNii Article

CiNii Books

researchmap

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

CiNii Article

researchmap

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

CiNii Article

researchmap

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

CiNii Article

researchmap

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

CiNii Article

researchmap

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

CiNii Article

researchmap

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。

DOI： 10.5458/bag.3.1_77

CiNii Article

CiNii Books

researchmap

その他リンク： http://agriknowledge.affrc.go.jp/RN/2010851804

記述言語：英語  

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.

DOI： 10.1271/bbb.130076

PubMed

researchmap

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.

PubMed

researchmap

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

CiNii Article

CiNii Books

researchmap

その他リンク： https://jlc.jst.go.jp/DN/JALC/00371501304?from=CiNii

記述言語：日本語   出版者・発行元：(一社)日本職業・災害医学会  

researchmap

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.

DOI： 10.1271/bbb.80889

Web of Science

PubMed

CiNii Article

researchmap

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.

DOI： 10.1271/bbb.80889

Web of Science

CiNii Article

researchmap

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.

DOI： 10.1177/039463200902200304

Scopus

PubMed

researchmap

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.

DOI： 10.1177/039463200902200107

Scopus

PubMed

researchmap

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.

DOI： 10.1177/039463200902200107

Scopus

PubMed

researchmap

記述言語：日本語   出版者・発行元：日本肺癌学会  

CiNii Article

CiNii Books

researchmap

記述言語：日本語   出版者・発行元：日本肺癌学会  

CiNii Article

CiNii Books

researchmap

記述言語：英語  

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.

DOI： 10.1016/j.envint.2008.01.003

PubMed

CiNii Article

researchmap

記述言語：英語  

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.

DOI： 10.1016/j.envint.2008.01.003

PubMed

CiNii Article

researchmap

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)

researchmap

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.

Web of Science

PubMed

researchmap

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.

Web of Science

researchmap

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  

Web of Science

researchmap

記述言語：英語  

200800

PubMed

researchmap

記述言語：日本語  

CiNii Article

CiNii Books

researchmap

記述言語：日本語  

CiNii Article

CiNii Books

researchmap

記述言語：日本語  

CiNii Article

CiNii Books

researchmap

記述言語：日本語  

CiNii Article

CiNii Books

researchmap

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

PubMed

CiNii Article

researchmap

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

PubMed

CiNii Article

researchmap

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

Web of Science

CiNii Article

researchmap

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

Web of Science

CiNii Article

researchmap

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

Web of Science

PubMed

CiNii Article

researchmap

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

Web of Science

PubMed

CiNii Article

researchmap

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

CiNii Article

CiNii Books

researchmap

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

記述言語：日本語   出版者・発行元：日本肺癌学会  

CiNii Article

CiNii Books

researchmap

記述言語：日本語   出版者・発行元：日本肺癌学会  

CiNii Article

CiNii Books

researchmap

記述言語：日本語   出版者・発行元：日本肺癌学会  

CiNii Article

CiNii Books

researchmap

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.

PubMed

researchmap

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.

PubMed

researchmap

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

Web of Science

PubMed

researchmap

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

PubMed

researchmap

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

PubMed

researchmap

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

Web of Science

PubMed

researchmap

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  

CiNii Article

CiNii Books

researchmap

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

researchmap

記述言語：日本語   出版者・発行元：日本職業・災害医学会  

CiNii Article

CiNii Books

researchmap

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

記述言語：日本語  

CiNii Article

CiNii Books

researchmap

記述言語：日本語  

CiNii Article

CiNii Books

researchmap

記述言語：日本語   出版者・発行元：日本衛生学会  

CiNii Article

CiNii Books

researchmap

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

記述言語：日本語  

CiNii Article

CiNii Books

researchmap

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

CiNii Article

CiNii Books

researchmap

その他リンク： http://search.jamas.or.jp/link/ui/2007192096

記述言語：英語  

Web of Science

researchmap

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.

Web of Science

researchmap

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.

DOI： 10.1271/bbb.60331

Web of Science

PubMed

CiNii Article

researchmap

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.

DOI： 10.1007/s10495-006-9235-4

PubMed

CiNii Article

researchmap

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.

DOI： 10.1271/bbb.60331

Web of Science

CiNii Article

researchmap

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.

DOI： 10.1007/s10495-006-9235-4

PubMed

CiNii Article

researchmap

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.

DOI： 10.1111/j.1365-2567.2006.02376.x

Web of Science

PubMed

researchmap

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.

DOI： 10.1111/j.1365-2567.2006.02376.x

Web of Science

PubMed

researchmap

researchmap

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.

DOI： 10.1271/bbb.50637

Web of Science

CiNii Article

researchmap

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.

DOI： 10.1271/bbb.50637

Web of Science

PubMed

CiNii Article

researchmap

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.

DOI： 10.1177/039463200601900409

Scopus

PubMed

researchmap

DOI： 10.1177/039463200601900409

Scopus

PubMed

researchmap

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

Web of Science

researchmap

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

Web of Science

PubMed

CiNii Article

researchmap

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

Web of Science

PubMed

CiNii Article

researchmap

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

Web of Science

PubMed

CiNii Article

researchmap

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

Web of Science

PubMed

CiNii Article

researchmap

200500

DOI： 10.4052/tigg.17.205

researchmap

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  

Web of Science

researchmap

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  

Web of Science

researchmap

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

Scopus

PubMed

researchmap

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

Scopus

PubMed

researchmap

researchmap

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

Scopus

PubMed

CiNii Article

researchmap

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

Scopus

PubMed

CiNii Article

researchmap

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

Scopus

PubMed

CiNii Article

researchmap

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

Scopus

PubMed

CiNii Article

researchmap

開催年月日： 2022年9月29日 - 2022年10月1日

会議種別：ポスター発表  

researchmap

開催年月日： 2022年9月29日 - 2022年10月1日

会議種別：ポスター発表  

researchmap

開催年月日： 2022年5月28日 - 2022年5月29日

researchmap

開催年月日： 2022年3月15日 - 2022年3月18日

researchmap

開催年月日： 2022年3月15日 - 2022年3月18日

researchmap

開催年月日： 2021年10月27日 - 2021年10月29日

researchmap

researchmap

会議種別：口頭発表（一般）  

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

会議種別：ポスター発表  

researchmap

researchmap

researchmap

researchmap

会議種別：ポスター発表  

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap

researchmap