Yuhki Yanase

Hiroshima University, Hirosima, Hiroshima, Japan

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Publications (37)128.99 Total impact

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    ABSTRACT: Toll-like receptor (TLR) 4 mediates inflammation and is also known to trigger apoptosis in microglia. Our time-lapse observations showed that lipopolysaccharide (LPS) stimulation induced rapid death in primary cultures of rat microglia, while a portion of the microglia escaped from death and survived for much longer than 2 days, in which time, all of the control cells had died. However, it remains unclear how the LPS-stimulated microglia subpopulation could continue to survive in the absence of any supplied growth factors. In the present study, to clarify the mechanism underlying the LPS-stimulated survival, we investigated whether microglia could produce their own survival factors in response to LPS, focusing on macrophage colony-stimulating factor (M-CSF), granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-34, which are mainly supplied by astrocytes or neurons. The LPS-stimulated microglia drastically induced the expression of the GM-CSF mRNA and protein, while M-CSF and IL-34 levels were unchanged. The surviving microglia also significantly upregulated the expression of GM-CSF receptor (GM-CSFR) mRNA without affecting M-CSFR. As for the GM-CSFR downstream signal, LPS resulted in the phosphorylation of STAT5 and its translocation to the nucleus in the surviving microglia. Moreover, a specific JAK2 inhibitor, NVP-BSK 805, suppressed STAT5 phosphorylation and microglia survival in response to LPS, indicating a critical role of the JAK2/STAT5 pathway in this survival mechanism. Together, these results suggest that a subpopulation of TLR4-activated microglia may survive by producing GM-CSF and up-regulating GM-CSFR. This autocrine GM-CSF pathway may activate the JAK2/STAT5 signaling pathway, which controls the transcription of survival-related genes. Finally, these surviving microglia may have neuroprotective functions because the neurons remained viable in co-cultures with these microglia.
    No preview · Article · Jan 2016 · Neurochemistry International
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    ABSTRACT: MGL_1304, a major allergen in human sweat for patients with atopic dermatitis and/or cholinergic urticaria, is secreted from Malassezia globosa on human skin. The amounts of MGL_1304 and IgE against MGL_1304 are evaluated by the histamine release test using basophils or mast cells sensitized with serum containing IgE against MGL_1304, and enzyme linked sorbent assay (ELISA) using MGL_1304 and anti-MGL_1304 antibodies. Here, we identified a human monoclonal IgE (ABS-IgE) that binds to the high affinity IgE receptor (FcεRI) and MGL_1304 with high affinity (KD=1.99nM) but does not release histamine from basophils and mast cells. An ELISA using ABS-IgE as a standard IgE revealed that the amount of IgE against MGL_1304 (1000 U/ml) in the standard sera of patients with AD, employed in our previous report, is 32 ng/ml. A sandwich ELISA using ABS-IgE as a detection antibody showed approximately 10 times lower detection limit for MGL_1304 than ELISA in which MGL_1304 is directly bound to an ELISA plate. Moreover, ABS-IgE prevented histamine release from mast cells and basophils by neutralizing MGL_1304 not only in a free form in solution, but also on FcεRI expressed on the cell surface without cell activation. ABS-IgE may be used both to quantify the amount of MGL_1304 and anti-MGL_1304 IgE, and possibly for the treatment of diseases caused/aggravated by type I allergy to MGL_1304.
    No preview · Article · Nov 2015 · Biochemical and Biophysical Research Communications

  • No preview · Article · Aug 2015 · Allergology International
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    ABSTRACT: We previously identified MGL_1304 secreted by Malassezia globosa as a sweat antigen for patients with atopic dermatitis (AD) and cholinergic urticaria (ChU). However, purifying native MGL_1304 from human sweat or culture supernatant of M. globosa (sup-MGL_1304) is costly and time-consuming. Moreover, recombinant MGL_1304 expressed by using Escherichia coli (TF-rMGL_1304) needs a large chaperon protein and lacks the original glycosylation of yeasts. Thus, we generated a recombinant MGL_1304 by Pichia pastoris (P-rMGL_1304) and investigated its characteristic features. Recombinant MGL_1304 proteins expressed by E. coli and P. pastoris were generated. Properties of these recombinants and native antigens were compared by western blot analysis, histamine release tests (HRT) of patients with AD and ChU, and β-hexosaminidase release tests with RBL-48 cells. P-rMGL_1304-specific IgE in sera of patients with AD were measured by sandwich ELISA. Western blot analysis revealed that IgE of patients with AD bound to all MGL_1304 recombinants and native antigens. The histamine releasing ability of P-rMGL_1304 was 100 times higher than that of TF-rMGL_1304, and was comparable to that of sup-MGL_1304. Degranulation rates of RBL-48 cells, sensitized with sera of patients with AD in response to the stimulation of P-rMGL_1304, were comparable to those of sup-MGL_1304, whereas those of TF-rMGL_1304 were relatively weak. The levels of P-rMGL_1304-specific IgE in sera of patients with AD were correlated with their disease severities. P-rMGL_1304 has an antigenicity comparable to the native antigen, and is more useful than TF-rMGL_1304, especially in HRT and degranulation assay of RBL-48 cells. Copyright © 2015 Japanese Society of Allergology. Production and hosting by Elsevier B.V. All rights reserved.
    Preview · Article · Apr 2015 · Allergology International
  • Y. Yanase · M. Hide
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    ABSTRACT: Noninvasive real-time evaluation of living cell conditions and functions is increasingly sought after in life science. Surface plasmon resonance (SPR) sensors can sensitively detect refractive index changes on the surface of a sensor chip without any labeling in a real-time manner. In this chapter, we first review the principle of SPR sensors and the applications of SPR sensors for detection of living cell reactions. We then introduce the technique to isolate and fix basophils on an SPR sensor chip. Finally, we describe the method to visualize individual basophil activation by means of SPR imaging (SPRI) sensor and the potential of basophil activation test by SPRI for clinical diagnosis of type I allergy.
    No preview · Article · Jan 2015 · Methods in Pharmacology and Toxicology
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    ABSTRACT: The identification of antigen that induces activation of mast cells and basophils by crosslinking IgE bound to the cell surface is crucial to avoid symptoms of allergic diseases. Surface plasmon resonance imaging (SPRI) possesses a great potential for clinical diagnosis of allergy, in that it reveals living cell activation following the binding of antigens to IgE, on real-time and single cell basis without artificial labeling. However, present technique of SPRI requires freshly isolated basophils of patients and cannot analyze multiple samples in parallel. To overcome such problems, we developed devices for SPRI to make a broad observation area and a multi-well SPRI sensor chip with a hydrophobic membrane. The employment of human IgE receptor-expressing mast cell lines (RBL-48 cells) sensitized with serum, collected and stored from less than a microliter of patient’s blood, allowed us to detect specific reactions of RBL-48 cells in response to antigens. This technique may be a useful tool as a high throughput screening system of type I allergy not only for freshly prepared basophils but also for sera stored in clinical practices.
    Preview · Article · Nov 2014 · Sensing and Bio-Sensing Research
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    ABSTRACT: Hereditary angioedema (HAE) presents as severe angioedema, which is mostly due to the C1 inhibitor (C1-INH) gene mutations. Environmental factors, minor trauma and oral contraceptives have been reported to induce angioedema attack, but the trigger may often be uncertain. Activated factor XII controlled by C1-INH facilitates bradykinin generation and also regulates coagulation cascade, but the relationship between edema formation and coagulation is still unclear. We have described a 35-year-old female patient with HAE, presenting with frequent angioedema attacks in the absence of an apparent triggering factor. She showed higher levels of FDP and D-dimer during angioedema than those in remission. In addition, tissue factor (TF), an initiator of the extrinsic coagulation cascade, was expressed on the surface of monocytes. It was significantly higher than that of monocytes from healthy controls and tends to further increase during attacks. The expression of TF on monocytes may play a role in the induction of angioedema attacks in HAE by activating the coagulation pathway in association with reduced functions of C1-INH.
    No preview · Article · Sep 2014 · The Journal of Dermatology
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    ABSTRACT: Immortal mast cell lines, such as RBL-2H3 and HMC-1 cells, are commonly utilized to investigate the function of mast cells. However, they are tumor cells carrying a gain-of-function mutation of Kit. We established an immortal mast cell line without Kit mutation, NCL-2, derived from NC mouse bone marrow. NCL-2 cells could be maintained without additional growth factors and thus could respond to exogenous growth signals. Moreover, NCL-2 cells expressed FcεRI and KIT, and release histamine and LTB4 in response to antigen stimulation. This cell line could be a useful tool to analyze proliferation, differentiation, and function of normal mast cells.
    Full-text · Article · Apr 2014 · FEBS Open Bio
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    ABSTRACT: Non-invasive real-time observations and the evaluation of living cell conditions and functions are increasingly demanded in life sciences. Surface plasmon resonance (SPR) sensors detect the refractive index (RI) changes on the surface of sensor chips in label-free and on a real-time basis. Using SPR sensors, we and other groups have developed techniques to evaluate living cells' reactions in response to stimuli without any labeling in a real-time manner. The SPR imaging (SPRI) system for living cells may visualize single cell reactions and has the potential to expand application of SPR cell sensing for clinical diagnosis, such as multi-array cell diagnostic systems and detection of malignant cells among normal cells in combination with rapid cell isolation techniques.
    Full-text · Article · Mar 2014 · Sensors
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    ABSTRACT: Background: MGL_1304 secreted by Malassezia globosa is contained in human sweat and induces histamine release from basophils in patients with atopic dermatitis (AD) at a high positive rate. The aims of this study were to establish the enzyme-linked immunosorbent assay (ELISA) measuring specific immunoglobulins against MGL_1304 and to investigate the levels of these immunoglobulins in sera of patients with various allergic diseases. Methods: Purified MGL_1304 from human sweat (QRX) and recombinant MGL_1304 (rMGL_1304) were prepared for ELISA. To quantify the amount of MGL_1304-specific immunoglobulins, the standard serum was created by pooling sera of 20 patients with AD whose basophils released histamine in response to QRX. A monoclonal antibody which exhibited the highest neutralizing ability against QRX was established as Smith-2, and used as a capture antibody for the assay of QRX-specific IgE. A total of 156 subjects [normal controls (n = 23), AD (n = 63), cholinergic urticaria (CU) (n = 24), bronchial asthma (n = 32), and allergic rhinitis (n = 14)] were enrolled in this study. Results: ELISA methods to quantify the specific IgE, IgG and IgG4 against MGL_1304 in sera were successfully established. Levels of QRX-specific IgE in sera of patients with AD and CU were significantly higher than those of normal controls. Moreover, the levels of QRX-specific IgE and rMGL_1304-specific IgE in patients with AD were significantly correlated with their disease severities. Conclusions: These ELISA methods to quantify the specific immunoglobulins against MGL_1304 are easy and useful means to assess allergy to MGL_1304. MGL_1304 contained in sweat is an important antigen for patients with AD and CU.
    Preview · Article · Jan 2014 · Allergology International
  • T. Obara · Y. Yanase · N. Kumazaki · T. Kawaguchi · M. Hide
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    ABSTRACT: We report a parallel surface plasmon resonance imaging (SPRI) system partially automated for functional analysis of living cells. To parallelize the assay, an array on 10 microfluidic chambers was placed in the view field of SPRI. We have shown the possibility of automated assay by demonstrating analysis of an experimental model of type I allergy. We stimulated sensitized rat basophilic leukemia cell line with the corresponding antigen in order to let them respond, and visualize the response of cells with the parallel SPRI system.
    No preview · Article · Jan 2014
  • K. Sakamoto · Y. Yanase · M. Hide · R. Miyake
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    ABSTRACT: We proposed a new allergic diagnostic chip applied with human basophils activation. Surface plasmon resonance (SPR) imaging biosensors using human basophils activation allows the allergic diagnosis with the less than 1 milliliter of peripheral blood [1] [2] [3]. However the isolation of basophilic leukocytes from small amount of blood sample is not easy because population rate of basophils in peripheral blood leukocytes is usually only 0.5%. In this presentation, we introduce the integrated chip of SPR imaging biosensors and micro-isolation column for those small amount of basophilic leukocytes. Then the isolation and detection performance of the integrated chip is checked.
    No preview · Article · Jan 2014
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    Full-text · Dataset · Nov 2013
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    ABSTRACT: Background: Non-steroidal anti-inflammatory drugs (NSAIDs), especially aspirin, and food additives (FAs) may exacerbate allergic symptoms in patients with chronic idiopathic urticaria and food-dependent exercise-induced anaphylaxis (FDEIA). Augmentation of histamine release from human mast cells and basophils by those substances is speculated to be the cause of exacerbated allergic symptoms. We sought to investigate the mechanism of action of aspirin on IgE-mediated histamine release. Methods: The effects of NSAIDs, FAs or cyclooxygenase (COX) inhibitors on histamine release from human basophils concentrated by gravity separation were evaluated. Results: Benzoate and tartrazine, which have no COX inhibitory activity, augmented histamine release from basophils similar to aspirin. In contrast, ibuprofen, meloxicam, FR122047 and NS-398, which have COX inhibitory activity, did not affect histamine release. These results indicate that the augmentation of histamine release by aspirin is not due to COX inhibition. It was observed that aspirin augmented histamine release from human basophils only when specifically activated by anti-IgE antibodies, but not by A23187 or formyl-methionyl-leucyl-phenylalanine. When the IgE receptor signaling pathway was activated, aspirin increased the phosphorylation of Syk. Moreover, patients with chronic urticaria and FDEIA tended to be more sensitive to aspirin as regards the augmentation of histamine release, compared with healthy controls. Conclusions: Aspirin enhanced histamine release from basophils via increased Syk kinase activation, and that the augmentation of histamine release by NSAIDs or FAs may be one possible cause of worsening symptoms in patients with chronic urticaria and FDEIA.
    Full-text · Article · Oct 2013 · Allergology International
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    ABSTRACT: BACKGROUND: Sweat is a major aggravating factor of atopic dermatitis (AD) and approximately 80% of patients with AD show type I hypersensitivity against sweat. OBJECTIVE: To identify and characterize an antigen in sweat that induces histamine release from basophils of patients with AD. METHODS: Basophil histamine-releasing activity in sweat was purified by a combination of chromatographies, and proteins were analyzed with mass spectrometry. Recombinant proteins of the sweat antigen were generated, and their biological characteristics were studied by immunoblots, histamine release tests, and neutralization assays. RESULTS: We identified a fungal protein, MGL_1304, derived from Malassezia globosa (M globosa) in the purified sweat antigen. Recombinant MGL_1304 induced histamine release from basophils of most of the patients with AD, in accordance with the semi-purified sweat antigen. Moreover, recombinant MGL_1304 abolished the binding of serum IgE of patients with AD to the semi-purified sweat antigen, or vice versa in immunoblot analysis, and attenuated the sensitization of RBL-48 mast cells expressing human FcεRI by serum IgE. Studies of truncated mutants of MGL_1304 indicated that IgE of patients with AD recognized the conformational structure of MGL_1304 rather than short peptide sequences. Western blot analysis of the whole lysate, the culture supernatant of M globosa, and the semi-purified sweat antigen showed that MGL_1304 was produced as a minor immunological antigen of M globosa with posttranslational modification, cleaved, and secreted as a 17-kDa major histamine-releasing sweat antigen. CONCLUSION: MGL_1304 is a major allergen in human sweat and could cause type I allergy in patients with AD.
    No preview · Article · May 2013 · The Journal of allergy and clinical immunology
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    ABSTRACT: A technique to visualize living cell activation in a real time manner without any labeling is required in the fields of life sciences and medicine. We have reported that surface plasmon resonance (SPR) sensors detect large changes of refractive index (RI) with living cells, such as mast cells, human basophils and lymphocytes. However conventional SPR sensors detect only an average change of RI with thousands of cells at detectable area on a sensor chip. Therefore, we developed an SPR imaging (SPRI) sensor with a CMOS camera and an objective lens in order to visualize RI of individual living cells and their changes upon stimuli. The SPRI sensor we developed could detect reactions of individual rat basophilic leukemia (RBL-2H3) cells and mouse keratinocyte cells in response to specific or nonspecific stimuli. Moreover, the sensor could detect the reactions of individual human basophils isolated from patients in response to antigens (allergens). Thus the technique can visualize the effect of various stimuli, inhibitors and/or conditions on cell reactions as change of intracellular RI distribution at single cell levels. Establishment of the technique to rapidly isolate cells from patient blood should enable us to utilize SPRI system as a high throughput screening system in clinical diagnosis, such as type I hypersensitivity and drug hypersensitivity, and as a tool to reveal novel phenomena in evanescent fields around plasma membranes.
    Full-text · Article · Feb 2013 · Allergology International

  • No preview · Article · Feb 2013 · Journal of Dermatological Science

  • No preview · Article · Feb 2013 · Journal of Dermatological Science
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    S Morioke · T Hiragun · Y Yanase · K Uchida · H Suzuki · K Iwamoto · M Hide
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    ABSTRACT: Immunoglobulin (Ig) E plays an important role in the pathogenesis of allergic diseases such as atopic dermatitis and allergic asthma.We previously reported that a sulfate polysaccharide, fucoidan, suppressed IgE production by murine B cells in vitro. However, the mechanism by which fucoidan suppresses IgE production remains unclear. We incorporated sulfate groups into cellulose and studied their biological characteristics in vitro to explore the possibility of converting biologically neutral polysaccharides to active reagents with antiallergic functions. Cellulose was chemically processed using N,N-dimethylformamide (DMF) and DMF-sulfurtrioxide and recovered as cellulose sulfate with a molecular weight of around 10 kDa. We then studied the effect of cellulose sulfate on IgE production from B cells, IgE class-switching, and populations of IgE-secreting B cells prepared from murine spleen. We also investigated the effects of sulfated cellulose on the production of interleukin (IL) 4 and interferon (IFN) gamma and the expression of T-bet mRNA by splenic T cells. The cytotoxicity of cellulose sulfate was also examined. Cellulose sulfate suppressed IgE production in B cells stimulated with IL-4 and anti-CD40 antibody by inhibiting IgE class-switch recombination and decreasing the number of IgE-secreting B cells in vitro. Moreover, both cellulose sulfate and fucoidan suppressed IL-4 production and enhanced IFN-gamma production by murine T cells stimulated with anti-CD3 and anti-CD28 antibodies, despite the decrease in T-bet mRNA expression. Cellulose gains an antiallergic effect on B cells and T cells with the addition of sulfate groups.
    Preview · Article · Jun 2012 · Journal of investigational allergology & clinical immunology: official organ of the International Association of Asthmology (INTERASMA) and Sociedad Latinoamericana de Alergia e Inmunología
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    ABSTRACT: We previously reported that about 80 % of patients with atopic dermatitis and 60 % with cholinergic urticaria revealed type I allergy against sweat, by means of skin test against autologous sweat and/or histamine-release test for peripheral blood basophils with semi-purified sweat antigen. In this study, we developed an assay for sera to neutralize histamine-releasing activity of semi-purified sweat antigen. The semi-purified sweat antigen was pre-incubated with serially diluted sera for 30 min at 37 °C and was subjected to histamine-release activity. Histamine release-neutralization (HRN) activities were calculated by measuring the amount of histamine release from basophils in the presence or absence of semi-purified sweat antigen. Of 62 subjects, 39 showed positive histamine release (≥5 %) from their basophils in response to semi-purified sweat antigen, and sera of 34 out of 39 subjects (87.2 %) were also positive in HRN activity (≥10 %). The specificity of the HRN assay was 0.522. Moreover, HRN activities in sera were largely correlated with degrees of histamine release from peripheral blood basophils of the same donors in response to sweat antigen. To identify the substance that neutralizes histamine-release activity, we removed IgE and IgG from the sera of HRN (+) subjects by column chromatography. The HRN activities in 30 out of 42 sera were largely reduced by the removal of IgG. On the other hand, sera of four subjects lost HRN activity by the removal of IgE, suggesting that the majority of HRN (+) subjects have serum IgG against the sweat antigen as well as IgE bound to peripheral basophils. Thus, the HRN assay maybe useful for the screening of type I allergy against sweat antigen.
    No preview · Article · Apr 2012 · Archives for Dermatological Research

Publication Stats

471 Citations
128.99 Total Impact Points

Institutions

  • 2005-2016
    • Hiroshima University
      • • Institute of Biomedical and Health Sciences
      • • Department of Dermatology
      • • Department of Molecular and Pathological Neuroscience
      • • Department of Pharmacology
      Hirosima, Hiroshima, Japan
  • 2013
    • Stony Brook University
      Stony Brook, New York, United States