IgE-Crosslinking-Induced Luciferase Expression Test as a Sensitive Indicator of Anisakis Allergy
Haruyo Akiyama; Masashi Niwa; Chisato Kurisaka; Yuto Hamada; Yuma Fukutomi; Reiko Teshima
Antibodies, Feb. 2025, [Reviewed]

Consideration of the EXiLE test for predicting anaphylaxis after diclofenac etalhyaluronate administration
Haruyo Akiyama; Chisato Kurisaka; Dai Muramatsu; Shuhei Takada; Kei Toyama; Keiji Yoshioka; Ryosuke Nakamura
Journal of Immunotoxicology, 03 Nov. 2024, [Reviewed]

Novel IgE crosslinking-induced luciferase expression method using human-rat chimeric IgE receptor-carrying mast cells.
Haruyo Akiyama; Chisato Kurisaka; Kenichi Kumasaka; Ryosuke Nakamura
Journal of immunological methods, Jun. 2024, [Reviewed]
BACKGROUND: The measurement of antigen-specific serum IgE is common in clinical assessments of type I allergies. However, the interaction between antigens and IgE won't invariably trigger mast cell activation. We previously developed the IgE crosslinking-induced luciferase expression (EXiLE) method using the RS-ATL8 mast cell line; however, the method may not be sensitive enough in some cases. METHODS: In this study, we introduced an NF-AT-regulated luciferase reporter gene into the RBL-2H3 rat mast cell line and expressed a chimeric high-affinity IgE receptor (FcεRI) α chain gene, comprising an extracellular domain from humans and transmembrane/intracellular domains from rats. RESULTS: We generated multiple clones expressing the chimeric receptor. Based on their responsiveness and proliferation, we selected the HuRa-40 clone. This cell line exhibited significantly elevated human α chain expression compared to RS-ATL8 cells, demonstrating a 10-fold enhancement of antigen-specific reactivity. Reproducibility across different batches and operators was excellent. Moreover, we observed a detectable response inhibition by an anti-allergy drugs (omalizumab and cyclosporin A). CONCLUSIONS: HuRa-40 cells-which carry the human-rat chimeric IgE receptor-comprise a valuable reporter cell line for the EXiLE method. Their versatility extends to various applications and facilitates high-throughput screening of anti-allergy drugs.

スギ花粉舌下免疫療法実施時におけるEXiLE法を用いた治療奏効性の評価
Dec. 2022, [Reviewed]

スギ花粉舌下免疫療法における奏効性予測法
Apr. 2021, [Reviewed]

[Effect of Cooking and Processing on Quantitation of Soybean Proteins].
Hiroko Watanabe; Haruyo Akiyama; Nobuhiko Osawa; Kaori Imura; Naomi Iseki; Sumiko Ueda; Tomoka Masaoka; Chie Akaboshi
Shokuhin eiseigaku zasshi. Journal of the Food Hygienic Society of Japan, 2021, [Reviewed]
We examined the effects of cooking and processing on the quantitation of soy protein in various soy-based foods. For the phosphate-buffered saline (PBS) extraction, the total protein content was measured by bicinchoninic acid assay, and the buffer extraction containing sodium dodecyl sulfate (SDS) and 2-mercaptoethanol (ME) was measured by the 2-D Quant Kit, and SDS-polyacrylamide gel electrophoresis analysis (SDS-PAGE) of each extraction was performed. Furthermore, measurements were performed by various ELISAs. During the tofu cooking process, the protein concentrations of soaked soybeans and Seigo (soybean homogenized with water) fluctuated- the change in protein solubility due to the amount of water during sample homogenization was considered to be a factor. It was thought that the decrease in protein concentration due to heating of Seigo during soymilk production was considered to indicate thermal denaturation of the protein, and that SDS and ME extraction in tofu may affect the measurement system. In cooking excluding roasted beans, proteins with a mass of 50 kDa or above and around 20 kDa were denatured, and in twice-fried tofu, proteins around 40 kDa were denatured, but the protein concentration excluding boiled soybeans did not decreased. Furthermore, the protein concentration from roasted beans, yuba, roasted okara and fried tofu increased with the cooking time, suggesting that the denaturation temperature of the protein shifted to a high temperature as the water content decreased. Both of the two types of ELISAs that comply with the official labeling system of foods containing allergenic substances were useful for detecting soybean protein by detecting proteins and peptides in processed soybean products, fermented foods excluding natto, and health foods.

Novel in vitro test for pollen-related vegetable/fruit allergy using the EXiLE method.
Haruyo Akiyama; Konatsu Kawamata; Yuma Fukutomi; Hiroshi Matsufuji; Shigemi Kai; Maki Miyazawa; Ryosuke Nakamura
Allergology international : official journal of the Japanese Society of Allergology, Jul. 2020, [Reviewed]

自己免疫・アレルギー疾患の難治化におけるマスト細胞の役割の解明　　　　　　　　　　　　　　　
Dec. 2019, [Reviewed]

Differentiation between control subjects and patients with chronic spontaneous urticaria based on the ability of anti-IgE autoantibodies (AAbs) to induce FcεRI crosslinking, as compared to anti-FcεRIα AAbs.
Satoshi Izaki; Shota Toyoshima; Takahiro Endo; Kazuko Kanegae; Satoshi Nunomura; Jun-Ichi Kashiwakura; Tomomi Sasaki-Sakamoto; Ryosuke Nakamura; Haruyo Akiyama; Chisei Ra; Koremasa Hayama; Tadashi Terui; Yoshimichi Okayama
Allergology international : official journal of the Japanese Society of Allergology, Jul. 2019, [Reviewed]
BACKGROUND: The reported prevalences of IgG autoantibodies (AAbs) to FcεRIα and IgE in sera from patients with chronic spontaneous urticaria (CSU) have varied, and these AAbs are also often observed in healthy control subjects. Regarding the histamine release activity of purified IgG from patients with CSU, the number of examined patients has been small. Thus, we sought to determine the prevalence and FcεRI crosslinking ability of these AAbs in a large number of patients with CSU and non-atopic control (NC) subjects. METHODS: We compared the concentrations of anti-IgE and anti-FcεRIα AAbs and the abilities of these AAbs to cause FcεRI aggregation in patients with CSU (n = 134) and NC subjects (n = 55) using ELISA and an in vitro elicitation test, respectively. RESULTS: The concentration of anti-IgE AAbs was significantly different between the NC subjects and the CSU patients (P < 0.0001, cutoff value: 0.558 μg/mL), whereas the concentration of anti-FcεRIα AAbs was not. A significant difference in the duration of illness was noted between patients with lower and those with higher concentrations of anti-IgE AAbs relative to the cutoff value. The abilities of anti-IgE AAbs, but not anti-FcεRIα AAbs, to induce FcεRI crosslinking were significantly higher in CSU patients than in NC subjects (P = 0.0106). CONCLUSIONS: In the Japanese population of CSU patients studied, the ability of the anti-IgE AAbs to induce FcεRI crosslinking differed significantly between NC subjects and CSU patients, suggesting the involvement of anti-IgE AAbs in the pathogenesis of CSU in the Japanese population.

「未病」の改善を目指した神奈川県衛生研究所の研究、教育、広報活動
Oct. 2017, [Reviewed]

神奈川県における過去10年間のアレルゲンを含む食品の検査結果について(平成18～27年度)
Sep. 2016, [Reviewed]

Fish collagen is an important panallergen in the Japanese population
Y. Kobayashi; H. Akiyama; J. Huge; H. Kubota; S. Chikazawa; T. Satoh; T. Miyake; H. Uhara; R. Okuyama; R. Nakagawara; M. Aihara; N. Hamada-Sato
Allergy, 08 Feb. 2016, [Reviewed]

Elevated receptor for activated C kinase 1 expression is involved in intracellular Ca²⁺ influx and potentially associated with compromised regulatory T cell function in patients with asthma
T. Negoro; S. Shimizu; M. Narushima; A. H. Banham; H. Wakabayashi; R. Takayanagi; T. Hagiwara; G. Roncador; T. Osabe; T. Yanai; M. Kin; K. Ikeda; A. Endo; H. Akiyama; Y. Nakano
Clinical & Experimental Allergy, 21 Aug. 2014, [Reviewed]
Summary

Background

Regulatory T cells (T_regs) are activated during anergy in response to T cell receptor (TCR) activation and functional immune suppression. Anergy of paediatric T_regs is partially dependent on intracellular calcium mobility; following TCR activation, T_regs do not exhibit increased intracellular Ca²⁺ concentration ([Ca²⁺]_i).

Objective

We determined whether [Ca²⁺]_i in adult T_regs defined their anergy, if intracellular Ca²⁺ movement was linked to regulatory functions, whether [Ca²⁺]_i was indicative of asthma pathology, and the potential molecular mechanism responsible for Ca²⁺ movement in T_regs.

Methods

T_regs were purified by the magnetic bead method, and their regulatory functions were assessed by monitoring carboxyfluorescein succinimidyl ester‐labelled responder T cell proliferation. The Ca²⁺ response of Fura‐2‐labelled cells was measured using a video image analysis system. To analyse the functions of T_regs at the molecular level, we generated Jurkat Tet‐On^® clones with doxycycline (Dox)‐induced forkhead box P3 (FOXP3) protein expression.

Results

CD4⁺CD25⁺CD127^−/low T_regs from participants without asthma did not elicit Ca²⁺ influx in response to TCR activation, exhibited little proliferation and suppressed proliferation of CD4⁺CD25⁻ T cells. In contrast, under similar conditions, T_regs from patients with asthma exhibited increased [Ca²⁺]_i and robust proliferation with partial loss of regulatory functions. FOXP3 protein levels in Tet‐On^® clones were high after both 2‐ and 5‐day Dox treatment; however, 5‐day cells were comparable with T_regs from patients with asthma, whereas 2‐day cells were similar to T_regs from participants without asthma. Increasing [Ca²⁺]_i induced a high level of receptor for activated C kinase 1 (RACK1) expression in 5‐day cells.

Conclusions and Clinical Relevance

We confirmed that T_regs in patients with asthma are functionally impaired and that the abnormal regulatory functions of these cells can be analysed by [Ca²⁺]_i following TCR engagement. Furthermore, the impaired functioning of T_regs evident in patients with asthma may be due to a high level of RACK1.

Plasminogen deficiency attenuates postnatal erythropoiesis in male C57BL/6 mice through decreased activity of the LH-testosterone axis.
Yurai Okaji; Yoshihiko Tashiro; Ismael Gritli; Chiemi Nishida; Aki Sato; Yoko Ueno; Sandra Del Canto Gonzalez; Makiko Ohki-Koizumi; Haruyo Akiyama; Hiromitsu Nakauchi; Koichi Hattori; Beate Heissig
Experimental hematology, Feb. 2012, [Reviewed]
Novel roles for the serine protease plasmin have been implicated recently in physiological and pathological processes. However, whether plasmin is involved in erythropoiesis is not known. In the present study, we studied the consequences of plasminogen deficiency on erythropoiesis in plasminogen-deficient (Plg knockout [KO]) mice. Erythroid differentiation was attenuated in male Plg KO mice and resulted in erythroblastic accumulation within the spleen and bone marrow, with increased apoptosis in the former, erythrocytosis, and splenomegaly, whereas similar erythropoietic defect was less prominent in female Plg KO mice. In addition, erythrocyte lifespan was shorter in both male and female Plg KO mice. Erythropoietin levels were compensatory increased in both male and female Plg KO mice, and resulted in a higher frequency of burst-forming units-erythroid within the spleen and bone marrow. Surprisingly, we found that male Plg KO mice, but not their female counterparts, exhibited normochromic normocytic anemia. The observed sex-linked erythropoietic defect was attributed to decreased serum testosterone levels in Plg KO mice as a consequence of impaired secretion of the pituitary luteinizing hormone (LH) under steady-state condition. Surgical castration causing testosterone deficiency and stimulating LH release attenuated erythroid differentiation and induced anemia in wild-type animals, but did not further decrease the hematocrit levels in Plg KO mice. In addition, complementation of LH using human choriogonadotropin, which increases testosterone production, improved the erythropoietic defect and anemia in Plg KO mice. The present results identify a novel role for plasmin in the hormonal regulation of postnatal erythropoiesis by the LH-testosterone axis.

Impaired Ca²⁺ regulation of CD4⁺CD25⁺ regulatory T cells from pediatric asthma.
Yoshiki Yamamoto; Takaharu Negoro; Akane Hoshi; Akiko Wakagi; Shunichi Shimizu; Alison H Banham; Masakazu Ishii; Haruyo Akiyama; Yuji Kiuchi; Susumu Sunaga; Takashi Tobe; Giovanna Roncador; Kazuo Itabashi; Yasuko Nakano
International archives of allergy and immunology, 2011, [Reviewed]
BACKGROUND: CD4(+)CD25(+) regulatory T (T(reg)) cells can control the allergic response to allergen, airway eosinophilia and airway hypersensitivity. We speculated that chronic inflammation persisting in asthma airways is dependent on abnormalities of these T(reg) cells. There are differences in the pathology of asthma in adults and children, and the airways of pediatric asthma are considered to be more naive than those of adults. Therefore, we analyzed the functionality of T(reg) cells in pediatric asthma and the relationship between T(reg) function and asthma symptoms. METHODS: The anergic state, which is one of the defining properties of T(reg), was analyzed by measuring intracellular Ca(2+) influx following T cell receptor (TCR) stimulation. FOXP3-positive cells and FOXP3 mRNA expression were measured by flow analysis and real-time PCR with the SYBR method, respectively. RESULTS: CD45RO(+) cells make up approximately 99% of CD4(+)CD25(high) T cells and 89% of CD4(+)CD25(low) T cells in human adult blood. The proportion of CD45RO(+) cells in CD4(+)CD25(+) (high + low) T cells from pediatric asthma was much smaller (about 56%). Interestingly, our data indicated that CD45RO(+) T(reg) cells from pediatric asthma aberrantly increased intracellular Ca(2+) concentrations following TCR activation compared with pediatric nonasthma controls. CONCLUSION: These impaired CD45RO(+) T(reg) cell functions were correlated with asthma symptoms. The correlation was observed in the group with a highly expressed atopic phenotype and longer duration of asthma. We suggest that chronic inflammation in pediatric asthma airways may be the result of impaired regulatory functions of CD45RO(+) T(reg) cells.

Tissue type plasminogen activator regulates myeloid-cell dependent neoangiogenesis during tissue regeneration.
Makiko Ohki; Yuichi Ohki; Makoto Ishihara; Chiemi Nishida; Yoshihiko Tashiro; Haruyo Akiyama; Hiromitsu Komiyama; Leif R Lund; Atsumi Nitta; Kiyofumi Yamada; Zhenping Zhu; Hideoki Ogawa; Hideo Yagita; Ko Okumura; Hiromitsu Nakauchi; Zena Werb; Beate Heissig; Koichi Hattori
Blood, 27 May 2010, [Reviewed]
Ischemia of the heart, brain, and limbs is a leading cause of morbidity and mortality worldwide. Treatment with tissue type plasminogen activator (tPA) can dissolve blood clots and can ameliorate the clinical outcome in ischemic diseases. But the underlying mechanism by which tPA improves ischemic tissue regeneration is not well understood. Bone marrow (BM)-derived myeloid cells facilitate angiogenesis during tissue regeneration. Here, we report that a serpin-resistant form of tPA by activating the extracellular proteases matrix metalloproteinase-9 and plasmin expands the myeloid cell pool and mobilizes CD45(+)CD11b(+) proangiogenic, myeloid cells, a process dependent on vascular endothelial growth factor-A (VEGF-A) and Kit ligand signaling. tPA improves the incorporation of CD11b(+) cells into ischemic tissues and increases expression of neoangiogenesis-related genes, including VEGF-A. Remarkably, transplantation of BM-derived tPA-mobilized CD11b(+) cells and VEGFR-1(+) cells, but not carrier-mobilized cells or CD11b(-) cells, accelerates neovascularization and ischemic tissue regeneration. Inhibition of VEGF signaling suppresses tPA-induced neovascularization in a model of hind limb ischemia. Thus, tPA mobilizes CD11b(+) cells from the BM and increases systemic and local (cellular) VEGF-A, which can locally promote angiogenesis during ischemic recovery. tPA might be useful to induce therapeutic revascularization in the growing field of regenerative medicine.

Participation of Th17 and Treg Cells in Pediatric Bronchial Asthma
Yoshiki Yamamoto; Takaharu Negoro; Akiko Wakagi; Akane Hoshi; Alison H. Banham; Giovanna Roncador; Haruyo Akiyama; Takashi Tobe; Susumu Sunaga; Yasuko Nakano; Kazuo Itabashi
Journal of Health Science, 2010, [Reviewed]

中毒原因物質(薬毒物)同定のためのデータベース作成　　　　　　　　　　　　　　　
Dec. 2009, [Reviewed]

生体内マトリックスメタロプロテイナーゼ-9の特異的活性抑制に伴う組織内好中球浸潤減少のG-CSF誘導血管新生における意義(Targeted deletion of matrix metalloproteinase-9 reduces neutrophil accumulation during G-CSF-induced neoangiogenesis)
佐藤 弥生; 大木 勇一; 秋山 晴代; ジャネット・ローゼンクヴィスト; イシマル・クリツリ; 奥村 康; 小川 秀興; 代田 浩之; ベアテ・ハイジッヒ; 服部 浩一; 大坂 顯通
Cytometry Research, Sep. 2009

The plasminogen fibrinolytic pathway is required for hematopoietic regeneration.
Beate Heissig; Leif R Lund; Haruyo Akiyama; Makiko Ohki; Yohei Morita; John Rømer; Hiromitsu Nakauchi; Ko Okumura; Hideoki Ogawa; Zena Werb; Keld Danø; Koichi Hattori
Cell stem cell, 13 Dec. 2007, [Reviewed]
Hematopoietic stem cells within the bone marrow exist in a quiescent state. They can differentiate and proliferate in response to hematopoietic stress (e.g., myelosuppression), thereby ensuring a well-regulated supply of mature and immature hematopoietic cells within the circulation. However, little is known about how this stress response is coordinated. Here, we show that plasminogen (Plg), a classical fibrinolytic factor, is a key player in controlling this stress response. Deletion of Plg in mice prevented hematopoietic stem cells from entering the cell cycle and undergoing multilineage differentiation after myelosuppression, leading to the death of the mice. Activation of Plg by administration of tissue-type plasminogen activator promoted matrix metalloproteinase-mediated release of Kit ligand from stromal cells, thereby promoting hematopoietic progenitor cell proliferation and differentiation. Thus, activation of the fibrinolytic cascade is a critical step in regulating the hematopoietic stress response.

Novel functions for a fibrinolytic pathway in controlling hematopoiesis
Koichi Hattori; Haruyo Akiyama; Leif R. Lund; Zena Werb; Beate Heissig
BLOOD, Nov. 2007, [Reviewed]

血管病変形成過程における骨髄由来炎症性細胞動員の意義　　　　　　　　　　　　　　　
Sep. 2007

Ablation of a peptidyl prolyl isomerase Pin1 from p53-null mice accelerated thymic hyperplasia by increasing the level of the intracellular form of Notch1.
K Takahashi; H Akiyama; K Shimazaki; C Uchida; H Akiyama-Okunuki; M Tomita; M Fukumoto; T Uchida
Oncogene, 31 May 2007, [Reviewed]
Tumor suppressor p53 is essential for checkpoint control in response to a variety of genotoxic stresses. DNA damage leads to phosphorylation on the Ser/Thr-Pro motifs of p53, which facilitates interaction with Pin1, a pSer/pThr-Pro-specific peptidyl prolyl isomerase. Pin1 is required for the timely activation of p53, resulting in apoptosis or cell cycle arrest. To investigate the physiological relationship between Pin1 and p53, we created Pin1-/-p53-/- mice. These p53-deficient mice spontaneously developed lymphomas, mainly of thymic origin, as well as generalized lymphoma infiltration into other organs, including the liver, kidneys and lungs. Ablation of Pin1, in addition to p53, accelerated the thymic hyperplasia, but the thymocytes in these Pin1-/-p53-/- mice did not infiltrate other organs. The thymocytes in 12-week-old Pin1-/-p53-/- mice were CD4(-)CD8(-) (double negative) and had significantly higher levels of the intracellular form of Notch1 (NIC) than the thymocytes of p53-/- or wild-type mice. Presenilin-1, a cleavage enzyme for NIC generation from full-length Notch1 was increased in the thymocytes of Pin1-/-p53-/- mice. Pin1 depletion also inhibited the degradation of NIC by proteasomes. These results suggest that both Pin1 and p53 control the normal proliferation and differentiation of thymocytes by regulating the NIC level.

ELISA method for monitoring human serum IgE specific for Cry1Ab introduced into genetically modified corn.
Osamu Nakajima; Reiko Teshima; Kayoko Takagi; Haruyo Okunuki; Jun-ichi Sawada
Regulatory toxicology and pharmacology : RTP, Feb. 2007, [Reviewed]
Enzyme-linked immunosorbent assay (ELISA) is the most convenient method of monitoring the occurrence of IgE antibodies specific for novel proteins in genetically modified (GM) foods. The levels of IgE specific for a recombinant protein, Cry1Ab, were determined using an ELISA method. A soluble form of the Cry1Ab protein purified from pCold1 vector-transformed Escherichia coli pTf16/BL21 was used as the ELISA coating antigen, and 1M NaCl was used as the washing buffer to remove IgE non-specifically bound to the coated antigen. Sera from 44 patients allergic to major food allergens were obtained, diluted 20-fold, tested, and found no identifiable IgE above background levels. We also tested sera from patients with corn allergy against whole extracts of non-GM and GM-corn (MON 810) using immunoblotting. The staining patterns were similar for the two types of corn. These results indicate that significant levels of IgE antibodies specific to Cry1Ab were not found in the sera of Japanese patients with food allergies.

Novel functions for a fibrinolytic pathway in controlling the stem cell niche.
Haruyo Akiyama; Leif R. Lund; Yohei Morita; Hiromitsu Nakauchi; Hideoki Ogawa; Ko Okumura; Keld Dano; Zena Werb; Beate Heissig; Koichi Hattori
BLOOD, Nov. 2006, [Reviewed]

A novel enzyme-linked immunosorbent assay specific for high-molecular-weight adiponectin.
Yasuko Nakano; Sachiko Tajima; Ai Yoshimi; Haruyo Akiyama; Motoo Tsushima; Toshihiro Tanioka; Takaharu Negoro; Motowo Tomita; Takashi Tobe
Journal of lipid research, Jul. 2006, [Reviewed]
Human plasma contains at least three forms of adiponectin: a trimer, a hexamer, and a high-molecular-weight (HMW) multimer. We purified HMW adiponectin from human plasma using its affinity to gelatin and obtained monoclonal antibodies against it. On Western blot analysis, the reactivity of these monoclonal antibodies was shown to be restricted to a non-heat-denatured form of adiponectin molecules. On heating, the collagen-like domain of adiponectin molecules became denatured, and thus the trimer form could not be maintained. From these, monoclonal antibodies against HMW adiponectin were suggested to react with the intact trimer of adiponectin. With these monoclonal antibodies, we developed a sandwich ELISA system for quantifying adiponectin in human serum. Its specificity was verified by analysis of serum fractions separated by gel-filtration chromatography, and our ELISA system was found to be HMW adiponectin-specific. With this novel ELISA, the HMW adiponectin concentrations were 8.4 +/- 5.5 microg/ml (mean +/- SD) in healthy women and 6.2 +/- 3.6 microg/ml in healthy men. Also, serum with a lower HMW adiponectin concentration was shown to have a lower HMW ratio (i.e., HMW adiponectin/total adiponectin).

Improved ELISA method for screening human antigen-specific IgE and its application for monitoring specific IgE for novel proteins in genetically modified foods.
Kayoko Takagi; Reiko Teshima; Osamu Nakajima; Haruyo Okunuki; Jun-ichi Sawada
Regulatory toxicology and pharmacology : RTP, Mar. 2006, [Reviewed]
For monitoring the occurrence of IgE antibody specific for novel proteins in genetically modified (GM) foods, ELISA is the most convenient method. The levels of IgE specific for recombinant proteins, phosphinothricin-N-acetyltransferase (PAT), CP4-EPSPS, and Cry9C were determined by ELISA using the sera from patients allergic to known allergens. Ovalbumin (OVA) and OVA-positive patient sera were used as positive control. In the ELISA, 20-fold-diluted sera tested were mostly negative for the specific IgE. However, the PAT-specific, but not CP4-EPSPS- or Cry9C-specific IgE in some patients was apparently higher than that of the healthy volunteers. To clarify the binding specificity of the antibody, we pre-incubated the sera with soluble PAT, but the inhibition was marginal, suggesting that the binding was non-specific. Therefore, we used 1M NaCl as a washing buffer to remove IgE non-specifically bound to the coated PAT. This washing step efficiently decreased non-specific binding. In contrast, OVA-specific IgE binding to OVA-coated plate was not affected by the washing. Finally, in this pilot study significant levels of IgE antibodies specific for the three proteins were not detected in the sera of Japanese food-allergy patients.

Effect of oral administration of CpG ODN-OVA on WBB6F1-W/Wv mice.
Reiko Teshima; Haruyo Okunuki; Yuji Sato; Hiroshi Akiyama; Tamio Maitani; Jun-ichi Sawada
Allergology international : official journal of the Japanese Society of Allergology, Mar. 2006, [Reviewed]
BACKGROUND: We have already reported that antigen-specific IgG1 antibody production in WBB6F1-W/Wv (W/Wv) mice after oral administration of ovalbumin (OVA) was extremely high. Active systemic anaphylaxis (ASA) was induced in these mice after intraperitoneal (i.p.) administration of OVA, and Th2-dominant helper T-cell activation occurred. In this study, we examined the effect of CpG oligodeoxynucleotide (ODN) conjugation of OVA on oral immunization of W/Wv mice. METHODS: W/Wv mice were sensitized by administration of 0.1 mg OVA or CpG ODN-OVA by gavage every day for 4 weeks, and the serum titers of OVA-specific IgG1, IgE, and IgG2a antibody were determined. ASA was induced by i.p. injection of OVA, and the changes in body temperature were monitored. In vitro production of Th1- and Th2- type cytokines by splenocytes re-stimulated with antigen was also measured. RESULTS: The antigen-specific IgG1 antibody titer in the CpG ODN-OVA-sensitized W/Wv mice was lower than in the OVA-sensitized group, but the IgG2a titer was higher. ASA was not induced by i.p. OVA challenge. There were significant increases in the production of Th1-type cytokine (IFN-gamma) by splenocytes in the CpG ODN-OVA-sensitized mice, but the Th2-type cytokine (IL-4) level in the splenocyte culture medium was lower. CONCLUSIONS: These results indicated that oral administration of CpG ODN-OVA conjugate significantly induced antigen-specific Th1 responses and reduced Th2 responses (allergic reactions) on re-stimulation. These findings suggest that CpG ODN-antigen conjugate may be useful as an oral vaccine.

Granulocyte colony-stimulating factor promotes neovascularization by releasing vascular endothelial growth factor from neutrophils.
Yuichi Ohki; Beate Heissig; Yayoi Sato; Haruyo Akiyama; Zhenping Zhu; Daniel J Hicklin; Kazunori Shimada; Hideoki Ogawa; Hiroyuki Daida; Koichi Hattori; Akimichi Ohsaka
FASEB journal : official publication of the Federation of American Societies for Experimental Biology, Dec. 2005, [Reviewed]
The granulocyte colony-stimulating factor (G-CSF) promotes angiogenesis. However, the exact mechanism is not known. We demonstrate that vascular endothelial growth factor (VEGF) was released by Gr-1+CD11b- neutrophils but not Gr-1-CD11b+ monocytes prestimulated with G-CSF in vitro and in vivo. Similarly, in vivo, concomitant with an increase in neutrophil numbers in circulation, G-CSF augmented plasma VEGF level in vivo. Local G-CSF administration into ischemic tissue increased capillary density and provided a functional vasculature and contributed to neovascularization of ischemic tissue. Blockade of the VEGF pathway abrogated G-CSF-induced angiogenesis. On the other hand, as we had shown previously, VEGF can induce endothelial progenitor cell (EPC) mobilization. Here, we show that G-CSF also augmented the number of circulating VEGF receptor-2 (VEGFR2) EPCs as compared with untreated controls. Blocking the VEGF/VEGFR1, but to a much lesser extent, the VEGF/VEGFR2 pathway in G-CSF-treated animals delayed tissue revascularization in a hindlimb model. These data clearly show that G-CSF modulates angiogenesis by increasing myelomonocytic cells (VEGFR1+ neutrophils) and their release of VEGF. Our results indicated that administration of G-CSF into ischemic tissue provides a novel and safe therapeutic strategy to improve neovascularization.

Low-dose irradiation promotes tissue revascularization through VEGF release from mast cells and MMP-9-mediated progenitor cell mobilization.
Beate Heissig; Shahin Rafii; Haruyo Akiyama; Yuichi Ohki; Yayoi Sato; Tejada Rafael; Zhenping Zhu; Daniel J Hicklin; Ko Okumura; Hideoki Ogawa; Zena Werb; Koichi Hattori
The Journal of experimental medicine, 19 Sep. 2005, [Reviewed]
Mast cells accumulate in tissues undergoing angiogenesis during tumor growth, wound healing, and tissue repair. Mast cells can secrete angiogenic factors such as vascular endothelial growth factor (VEGF). Ionizing irradiation has also been shown to have angiogenic potential in malignant and nonmalignant diseases. We observed that low-dose irradiation fosters mast cell-dependent vascular regeneration in a limb ischemia model. Irradiation promoted VEGF production by mast cells in a matrix metalloproteinase-9 (MMP-9)-dependent manner. Irradiation, through MMP-9 up-regulated by VEGF in stromal and endothelial cells, induced the release of Kit-ligand (KitL). Irradiation-induced VEGF promoted migration of mast cells from the bone marrow to the ischemic site. Irradiation-mediated release of KitL and VEGF was impaired in MMP-9-deficient mice, resulting in a reduced number of tissue mast cells and delayed vessel formation in the ischemic limb. Irradiation-induced vasculogenesis was abrogated in mice deficient in mast cells (steel mutant, Sl/Sl(d) mice) and in mice in which the VEGF pathway was blocked. Irradiation did not induce progenitor mobilization in Sl/Sl(d) mice. We conclude that increased recruitment and activation of mast cells following irradiation alters the ischemic microenvironment and promotes vascular regeneration in an ischemia model. These data show a novel mechanism of neovascularization and suggest that low-dose irradiation may be used for therapeutic angiogenesis to augment vasculogenesis in ischemic tissues.

【幹細胞生物学の新たな展開】幹細胞制御 造血幹細胞のMobilizationとHomingの制御機構
Aug. 2005, [Invited]

細胞の遊走の制御・ニッシェ 骨髄Nicheからの細胞動員と組織再生機構　　　　　　　　　　　　　　　
Jul. 2005

Gene expression profiling of dexamethasone-treated RBL-2H3 cells: induction of anti-inflammatory molecules.
Ryosuke Nakamura; Haruyo Okunuki; Seiichi Ishida; Yoshiro Saito; Reiko Teshima; Jun-Ichi Sawada
Immunology letters, 15 May 2005, [Reviewed]
Glucocorticoids are well known for their anti-inflammatory effect through the regulation of gene expression in many types of immune cells, including mast cells. However, the genes that are involved in suppression of mast cell-mediated inflammation by glucocorticoids have not been fully identified. Therefore, we examined the dexamethasone (Dex)-responsive genes in RBL-2H3 mast cells using a high-density oligonucleotide microarray technique. Gene expression profiling revealed that the antigen-induced up-regulation of pro-inflammatory factors, including monocyte chemoattractant protein-1, was markedly inhibited by 100 nM Dex. On the other hand, Dex treatment itself caused the substantial up-regulation of many genes, including phenylethanolamine-N-methyl transferase (PNMT) and cytokine-inducible SH2-containing protein (CISH), in the mast cells. The expression of these two genes significantly increased 6 h after Dex exposure and lasted for more than 24 h. Considering that PNMT is the rate-determining enzyme in epinephrine synthesis and that CISH is a suppressor of cytokine signaling, these Dex-responsive genes may be potential anti-inflammatory factors. Thus, gene expression profiling suggested that Dex might exert its anti-inflammatory effect through two pathways in mast cells: the suppression and induction of potentially pro- and anti-inflammatory factors, respectively.

The hyperresponsiveness of W/W(v) mice to oral sensitization is associated with a decrease in TCRgammadelta-T cells.
Haruyo Okunuki; Reiko Teshima; Yuji Sato; Ryosuke Nakamura; Hiroshi Akiyama; Tamio Maitani; Jun-Ichi Sawada
Biological & pharmaceutical bulletin, Apr. 2005, [Reviewed]
We have already reported that WBB6F1-W/W(v) (W/W(v)) mice, which have mutations in the c-kit gene, are highly susceptible to oral sensitization, and that the proportion of TCRgammadelta-T cells among the intraepithelial lymphocytes (IELs) (gammadelta-IELs) of W/W(v) is much lower than in congenic wild-type (+/+) mice. In this study we examined an inhibitory role of gammadelta-IELs in oral sensitization using two different methods. First, wild-type (+/+) mice were sensitized by oral administration of 1.0 mg ovalbumin (OVA) by gavage every day for 9 weeks after anti-TCRgammadelta antibody treatment 4 times. The treatment resulted in an enhanced OVA-specific IgG1 antibody production, active systemic anaphylaxis (ASA), and Th2-dominant cytokine production. Next, W/W(v) mice whose bone marrow cells were reconstituted from C57BL/6J mice for 5 months were sensitized by oral administration of OVA. The OVA-specific IgG1 antibody titer in the bone marrow-reconstituted W/W(v) mice was neither significantly enhanced, nor ASA was induced. The proportion of gammadelta-IELs in the reconstituted mice was much higher than that in the untreated W/W(v) mice. The above findings suggest that the decrease or increase in number of gammadelta-IELs enhances or decreases oral sensitization respectively. These results show that gammadelta-IELs have an important role in the oral tolerance to food antigens.

Survey of Food and Airborne Allergen-specific IgE Levels in a General Population of 3-year-old Japanese Children
Kayoko Takagi; Reiko Teshima; Haruyo Okunuki; Akiko Hachisuka; Jun-ichi Sawada; Kohichi Kojima; Kazuko Takahashi; Motoyasu Ohsawa; Takahiko Yoshida
Allergology International, 2005, [Reviewed]

Kinetic analysis of pepsin digestion of chicken egg white ovomucoid and allergenic potential of pepsin fragments.
Kayoko Takagi; Reiko Teshima; Haruyo Okunuki; Satsuki Itoh; Nana Kawasaki; Toru Kawanishi; Takao Hayakawa; Yoichi Kohno; Atsuo Urisu; Jun-Ichi Sawada
International archives of allergy and immunology, Jan. 2005, [Reviewed]
BACKGROUND: The allergenic potential of chicken egg white ovomucoid (OVM) is thought to depend on its stability to heat treatment and digestion. Pepsin-digested fragments have been speculated to continue to exert an allergenic potential. OVM was digested in simulated gastric fluid (SGF) to examine the reactivity of the resulting fragments to IgE in sera from allergic patients. METHODS: OVM was digested in SGF and subjected to SDS-PAGE. The detected fragments were then subjected to N-terminal sequencing and liquid chromatography/mass spectrometry/mass spectrometry analysis to confirm the cleavage sites and partial amino acid sequences. The reactivity of the fragments to IgE antibodies in serum samples from patients allergic to egg white was then determined using Western blotting (n=24). RESULTS: The rate of OVM digestion depended on the pepsin/OVM ratio in the SGF. OVM was first cleaved near the end of the first domain, and the resulting fragments were then further digested into smaller fragments. In the Western blot analysis, 93% of the OVM-reactive sera also bound to the 23.5- to 28.5-kDa fragments, and 21% reacted with the smaller 7- and 4.5-kDa fragments. CONCLUSION: When the digestion of OVM in SGF was kinetically analyzed, 21% of the examined patients retained their IgE-binding capacity to the small 4.5-kDa fragment. Patients with a positive reaction to this small peptide fragment were thought to be unlikely to outgrow their egg white allergy. The combination of SGF-digestibility studies and human IgE-binding experiments seems to be useful for the elucidation and diagnosis of the allergenic potential of OVM.

3歳児の食物並びに吸入アレルゲン特異的IgE抗体の実態調査
Sep. 2004, [Reviewed]

【食物アレルギーを科学する】食物アレルギーの動物モデル　　　　　　　　　　　　　　　
Jul. 2004, [Reviewed]

Method for Delivering Radiolabeled Single-Chain Fv Antibody to the Brain
Nakajima Osamu; Hachisuka Akiko; Okunuki Haruyo; Takagi Kayoko; Teshima Reiko; Sawada Jun-ichi
Journal of Health Science, 2004, [Reviewed]

Oral sensitization of W/W(v) mice with ovalbumin and possible involvement of the decrease in gammadelta-T cells.
Haruyo Okunuki; Reiko Teshima; Naoki Harikai; Shinobu Sakai; Hiroshi Akiyama; Tamio Maitani; Jun-Ichi Sawada
Biological & pharmaceutical bulletin, Sep. 2003, [Reviewed]
Mast-cell-deficient WBB6F1-W/W(v) mice (W/W(v)) and congenic wild-type (+/+) mice were sensitized by oral administration of 0.1 or 1.0 mg ovalbumin (OVA) in the form of gavage every day for 9 weeks, and active systemic anaphylaxis (ASA) was induced by intraperitoneal injection of OVA. Production of OVA-specific IgG1 in response to oral sensitization of the W/W(v) mice was very high, and the production of IL-4, IL-5 and IL-10 by splenocytes re-stimulated with OVA in vitro was increased. These findings suggest that Th2-dominant helper T-cell activation had occurred. By contrast, production of OVA-specific IgG1 was low in +/+ mice, and no significant increase in production of Th2-type cytokines by the splenocytes of +/+ mice was observed. Population analysis in Peyer's patches by flow cytometry revealed that the proportion of the CD11c(+) cell in the W/W(v) mice was slightly increased after antigen stimulation. Analysis of the cell surface markers of intraepithelial lymphocytes (IELs) by flow cytometry showed that the proportion of TCRgammadelta-T cells was extremely lower in the W/W(v) mice, especially in the antigen sensitized group. The proportion of TCRgammadelta-T cells in the splenocytes of W/W(v) mice was also lower than in +/+ mice. Taken together, the above findings indicate that W/W(v) mice seems to be a good model not only for studying the induction mechanism of food allergy but for examining the role of TCRgammadelta-T cells in food-induced hypersensitivity.

Comparative study of in vitro digestibility of food proteins and effect of preheating on the digestion.
Kayoko Takagi; Reiko Teshima; Haruyo Okunuki; Jun-ichi Sawada
Biological & pharmaceutical bulletin, Jul. 2003, [Reviewed]
Information on the comparative digestibility of food allergens and non-allergenic proteins is crucial when stability to digestion is to be used as a criterion to assess the allergenic potential of novel proteins. Preheating effect on in vitro digestibility has not been fully examined. In this study we investigated the preheating effect of in vitro digestibility of several proteins and their proteolytic fragments in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF). Five major food allergens, ovalbumin (OVA), ovomucoid (OVM), beta-lactoglobulin (BLG), bovine serum albumin (BSA), soybean trypsin inhibitor (STI), four proteins of unproven allergenicity, horseradish peroxidase (HRP), ribulose-1,5-bisphosphate carboxylase/oxidase (RBC), phosphinothricin acetyltransferase (PAT) and zein from corn, and plant lectin, concanavalin A (Con A) were preheated (at 100 degrees C for 5 min) or not preheated, and then digested in SGF or SIF. Food allergens were relatively stable in both SGF and SIF. Among the allergens, digestibility of OVA in both SGF and SIF was markedly decreased, and BLG and STI were relatively stable after preheating. Digestibility of ConA in SGF and SIF was markedly decreased by preheating. Digestibility of non-allergenic proteins in SGF was higher than the allergenic proteins. From these results, because of the marked increase of the digestibility in several proteins by preheating, systematic information concerning the effect of food treatment on protein digestion is necessary to assess the relationship between allergenic potential and the digestibility of food protein.

Determination of enzymatic activity of 5-enolpyruvylshikimate-3-phosphate synthase by LC/MS.
Haruyo Okunuki; Hiroshi Akiyama; Reiko Teshima; Akihiro Hino; Yukihiro Goda; Jun-ichi Sawada; Masatake Toyoda; Tamio Maitani
Shokuhin eiseigaku zasshi. Journal of the Food Hygienic Society of Japan, Apr. 2003, [Reviewed]
A liquid chromatography-mass spectrometry (LC/MS) method for determining the enzymatic activity of 5-enolpyruvylshikimate-3-phosphate synthase (EPSP synthase), an enzyme of the shikimate pathway, was developed. EPSP synthase catalyzes the formation of 5-enolpyruvylshikimate-3-phosphate (EPSP) from shikimate-3-phosphate (S-3-P) and phosphoenolpyruvate (PEP) in microorganisms and plants. The enzymatic activity of EPSP synthase was assessed by the determination of EPSP after a 30-min incubation with S-3-P and PEP using the LC/MS system. EPSP synthase activity is given in terms of the produced EPSP (pmol/min/mg protein). Glyphosate (N-phosphonomethyl glycine)-tolerant EPSP synthase from the Agrobacterium sp. strain CP4 (CP4-EPSP synthase) in genetically modified soybeans (GM-soybeans) was found to have an enzymatic activity of 736 EPSP pmol/min/mg protein in the presence of 3 nmol of S-3-P. In contrast, the enzyme activity of non-GM-soybeans was 21 EPSP pmol/min/mg protein. The EPSP synthase activity was markedly decreased in the non-GM-soybeans by the addition of glyphosate, but the enzyme activity of the GM-soybeans was only slightly decreased with this treatment. This LC/MS system could also be applicable to the measurement of EPSP synthase activity in different plant species and the detection of herbicide-tolerant EPSP synthase in GM foods.

Gene expression profiling of Ca2+-ATPase inhibitor DTBHQ and antigen-stimulated RBL-2H3 mast cells
R. Nakamura; S. Ishida; S. Ozawa; Y. Saito; H. Okunuki; R. Teshima; J. Sawada
Inflammation Research, Dec. 2002, [Reviewed]

Effect of subchronic feeding of genetically modified corn (CBH351) on immune system in BN rats and B10A mice.
Reiko Teshima; Takahiro Watanabe; Haruyo Okunuki; Kazuto Isuzugawa; Hiroshi Akiyama; Hiroshi Onodera; Toshio Imai; Masatake Toyoda; Jun-ichi Sawada
Shokuhin eiseigaku zasshi. Journal of the Food Hygienic Society of Japan, Oct. 2002, [Reviewed]
Subchronic animal feeding studies to examine the effect on the immune system of genetically modified corn CBH351, which contains the Cry9C protein derived from Bacillus thuringiensis subspecies tolworthi, were conducted in female BN rats and B10A mice. The studies were designed to compare the effect of a line of genetically modified corn CBH351 (GM corn) with that of isoline corn (non-GM corn). Heat-treated corn meal was incorporated into the diets of the rats and mice at a concentration of 50%. The study duration was 13 weeks. Growth, food intake, and organ weights of the thymus, spleen, and liver were compared between animals fed the non-GM and GM lines. The histological findings in thymus, spleen, mesenteric lymph nodes, Peyer's patches, small intestines, liver, kidney, and bone marrow, and the presence of Cry9C-specific IgE, IgG, IgG1 and IgA antibodies in serum were also compared. The results showed no significant differences in growth, feeding value, or the histological findings in immunity-related organs between the animals fed the GM and non-GM lines. Production of Cry9 C-specific IgE and IgA was not detected in the serum of either group. Production of Cry9C-specific IgG and IgG1 was slightly increased in the 50% GM groups of BN rats. No Cry9C-specific IgG or IgG1 was detected in the serum of BN rats fed the diet containing 5% GM-corn In conclusion, no immunotoxic activity was detected in the GM-corn-fed rats and mice in this subchronic dietary study.

Increased digestibility of two products in genetically modified food (CP4-EPSPS and Cry1Ab) after preheating.
Haruyo Okunuki; Reiko Teshima; Teruko Shigeta; Jun-ichiro Sakushima; Hiroshi Akiyama; Yukihiro Goda; Masatake Toyoda; Jun-ichi Sawada
Shokuhin eiseigaku zasshi. Journal of the Food Hygienic Society of Japan, Apr. 2002, [Reviewed]
We performed experiments on in vitro digestion of newly expressed proteins by SGF (simulated gastric fluid) and SIF (simulated intestinal fluid) to assess the allergenicity of food components derived from biotechnological modification. For newly expressed proteins, we chose CP4-EPSPS (5-enolpyruvylshikimate-3-phosphate synthase from Agrobacterium sp. strain CP4) and Cry1Ab derived from Bacillus thuringiensis subsp. kurstaki strain HD-1. The former is expressed in GM-soybeans and the latter is expressed in GM-corns. Firstly, we examined the digestibility of purified CP4-EPSPS and Cry1Ab by SGF. Both proteins were rapidly digested within 60 sec. After preheating, the digestibility by SGF was slightly increased. Secondly, CP4-EPSPS in GM-soybean extracts and Cry1Ab in GM-corn extracts were digested by SGF. The digestion time of both proteins by SGF was almost the same as that of the purified proteins. Thirdly, the digestibility of CP4-EPSPS and Cry1Ab by SIF was examined. The digestion time of these proteins was 240 min or more. However, digestibility of these proteins by SIF was dramatically increased by preheating, and the digestion time was less than 5 sec. Fourthly, CP4-EPSPS in GM-soybean extracts and Cry1Ab in GM-corn extracts were digested by SIF. Digestion time of both proteins by SIF was almost the same as that of the purified proteins. From these results, we concluded that the digestibility of both CP4-EPSPS and Cry1Ab by SGF and SIF was increased by preheating. Therefore, we suggest that the allergenicity of both proteins should be extremely low because of the easy digestibility of these proteins by SGF and also by SIF with preheating.

Examination of oral sensitization with ovalbumin in Brown Norway rats and three strains of mice
Hiroshi Akiyama; Reiko Teshima; Jun-ichiro Sakushima; Haruyo Okunuki; Yukihiro Goda; Jun-ichi Sawada; Masatake Toyoda
Immunology Letters, Aug. 2001, [Reviewed]

Induction of active systemic anaphylaxis by oral sensitization with ovalbumin in mast-cell-deficient mice
Haruyo Okunuki; Reiko Teshima; Jun-ichiro Sakushima; Hiroshi Akiyama; Yukihiro Goda; Masatake Toyoda; Jun-ichi Sawada
Immunology Letters, Nov. 2000, [Reviewed]

遺伝子組換え,非組換え大豆のマウス,ラットへの混餌投与による免疫系への影響
手島 玲子; 穐山 浩; 奥貫 晴代; 佐久嶋 順一郎; 合田 幸広; 小野寺 博志; 澤田 純一; 豊田 正武
食品衛生学雑誌, Jun. 2000, [Reviewed]

Effect of Ca2+ ATPase inhibitors on MCP-1 release from bone marrow-derived mast cells and the involvement of p38 MAP kinase activation
R Teshima; J Onose; H Okunuki; J Sawada
INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY, Jan. 2000, [Reviewed]

マスト細胞MCP-1のイムノアッセイ及びRT-PCRによる定量
2000, [Invited]

Construction of an Expression System of Shikimate Kinase II and Preparation of Shikimic Acid 3-Phosphate.
食品衛生学雑誌, 1999, [Reviewed]

舌下免疫療法実施時における血清中中和抗体活性のin vitro評価法の開発　　　　　　　　　　　　　　　
May 2018

バイオテクノロジーを用いて得られた食品のリスク管理及び国民受容に関する研究 バイオテクノロジー応用食品のプロテオーム解析　　　　　　　　　　　　　　　
2017

Novel in vitro test for oral allergy syndrome using the EXiLE method　　　　　　　　　　　　　　　
秋山晴代; 中村亮介; 福冨友馬; 栗原和幸; 中野泰子; 根来孝治; 宮澤眞紀
2016

バイオマーカーを標的とした非定型抗精神病薬による糖尿病発症機序の解明及び発症予測に関する研究　　　　　　　　　　　　　　　
2011

EXiLE法におけるRS-ATL8細胞とHuRa-40細胞の比較 長芋アレルギーでの検討　　　　　　　　　　　　　　　
Sep. 2024

IgEの架橋活性を指標とする新規アレルギー試験法の開発　　　　　　　　　　　　　　　
Sep. 2024, [Invited]

Correlation of treatment response to Japanese cedar pollen SLIT and EC50 of EXiLE response　　　　　　　　　　　　　　　
栗坂知里; 菅原健; 蛯名梨音; 齊藤美優; 中村亮介; 土井雅津代; 秋山晴代
日本薬学会年会要旨集(Web), 2024

Development of in vitro allergy testing method using saliva　　　　　　　　　　　　　　　
秋山晴代; 栗坂知里; 小川裕子; 大橋知子; 皆川範亨; 原田佳英; 渡部明日香; 池本守; 中村亮介; 矢ノ下良平
日本薬学会年会要旨集(Web), 2024

免疫療法・薬物療法(基礎) EXiLE法を用いたスギ花粉舌下免疫療法における抗IgE因子の評価
Aug. 2023

Development of high-throughput screening method for environmental allergy exacerbating substances by evaluation of human IgE cross-linking activity　　　　　　　　　　　　　　　
秋山晴代; 栗坂知里; 渡部明日香; 大橋知子; 原田佳英; 中村亮介
日本免疫毒性学会学術年会講演要旨集, 2023

Establishment of high-throughput screening system to evaluate IgE cross-linking activity.　　　　　　　　　　　　　　　
秋山晴代; 栗坂知里; 石井巧; 佐々木彩夢; 原有彩; 中村亮介
日本薬学会年会要旨集(Web), 2023

高感度I型アレルギー試験法の開発を目的としたIgEキメラ受容体発現細胞の作製　　　　　　　　　　　　　　　
Mar. 2022

Development of highly-sensitive allergy test using chimeric IgE receptor-expressing cultured mast cells　　　　　　　　　　　　　　　
秋山晴代; 栗坂知里; 田所哲; 熊坂謙一; 中村亮介
日本免疫毒性学会学術年会講演要旨集, 2022

ヒトIgE受容体発現新規細胞株HuRa-40細胞の機能性評価　　　　　　　　　　　　　　　
2022

コロナ禍におけるセミナー科目のオンライン化対応と授業効果測定の検証　　　　　　　　　　　　　　　
Mar. 2021

食物アレルゲンの架橋活性に及ぼすマイクロプラスチックの影響　　　　　　　　　　　　　　　
2021

加熱・加圧処理による魚肉低アレルゲン化の検討 EXiLE法による応答性評価
Oct. 2020

加熱・加圧処理による魚肉アレルゲンの変化 魚アレルギー患者血清を用いたEXiLE法による評価
Sep. 2020

EXiLE法を用いた加熱・加圧処理による魚肉アレルゲン性低減化の評価　　　　　　　　　　　　　　　
Mar. 2020

加熱・加圧処理による魚肉アレルゲンの変化:魚アレルギー患者血清を用いたEXiLE法による評価　　　　　　　　　　　　　　　
2020

The validation of EXiLE test for House dust mite allergy.　　　　　　　　　　　　　　　
Saori Ohmae; Ryosuke Nakamura; Haruyo Akiyama; Katsuyo Ohashi-Doi
The 68th Annual Meeting of Japanese Society of Allergology, Jun. 2019

免疫療法 スギ花粉症舌下免疫療法の奏効性は血清中特異的IgEの架橋能と相関する
May 2019

スギ花粉症の舌下免疫療法の奏効性は血清中IgEの架橋能と相関する　　　　　　　　　　　　　　　
Mar. 2019

スギ花粉舌下免疫療法実施時における減感作状態のin vitro評価(第2報)　　　　　　　　　　　　　　　
2019

花粉-食物アレルギー症候群におけるEXiLE法を用いた交差反応性評価　　　　　　　　　　　　　　　
2019

Crosslinking ability of the serum IgE correlates to the effectiveness of allergen immunotherapy for Japanese cedar pollen.　　　　　　　　　　　　　　　
Nakamura, R.; Akiyama, H.; Sakurai, D.; Matsuzawa, Y.; Saito, Y.; Okamoto, Y.
WAO International Scientific Conference 2018, Dec. 2018

舌下免疫療法実施時における血清中中和抗体活性のin vitro評価法の開発
May 2018

経口免疫寛容誘導時における減感作状態のEXiLE法を用いた評価　　　　　　　　　　　　　　　
Mar. 2018

スギ花粉舌下免疫療法実施時における減感作状態のin vitro評価法の検討　　　　　　　　　　　　　　　
2018

管内宿泊施設における食物アレルギー対策の実態調査
Nov. 2017

未病教育広報活動が未病認知と理解拡大に与えた効果の検証
Nov. 2017

「未病」の改善を目指した神奈川県衛生研究所の研究、教育、広報活動
Oct. 2017

乳幼児気道評価法の新しい展望 喘息患者における各FOXP3バリアント発現とその機能との関係
May 2017

抗原・免疫療法(基礎) EXiLE法を用いた経口免疫療法実施時における減感作状態の評価法の検討(第2報)
May 2017

「未病」の認知と理解拡大を目指した教育広報活動充実への研究成果活用とその効果の検証　　　　　　　　　　　　　　　
Mar. 2017

アディポネクチンアンチセンス、センストランスジェニックマウスの表現型解析　　　　　　　　　　　　　　　
Sep. 2016

神奈川県における過去10年間のアレルゲンを含む食品の検査結果について(平成18～27年度)
Sep. 2016

各種大豆加工食品のアレルゲン性評価法 加工によるアレルゲンの変性
May 2016

各種大豆加工食品のアレルゲン性評価法 EXiLE法を用いた加工に伴う抗原性の変化の解析
May 2016

易炎症性マウスを用いた未病に関与する診断ターゲットの探索　　　　　　　　　　　　　　　
Mar. 2016

EXiLE法は口腔アレルギー症候群の新しいin vitro検査法として有用である　　　　　　　　　　　　　　　
Oct. 2015

EXiLE法を用いた経口免疫療法実施時における減感作状態の評価法の検討
Apr. 2015

口腔アレルギー症候群における新たなin vitro試験法の検討　　　　　　　　　　　　　　　
Mar. 2015

EXiLE法を用いた口腔アレルギー症候群の新たなin vitro検査法の検討　　　　　　　　　　　　　　　
2014

関節リウマチにおけるアディポネクチンの生理的機能解明　　　　　　　　　　　　　　　
2012

非定型抗精神病薬の糖尿病病態発症機構の解明　　　　　　　　　　　　　　　
Sep. 2011

非定型抗精神病薬の糖尿病病態発症機構の解明　　　　　　　　　　　　　　　
Dec. 2010

関節リウマチにおけるアディポネクチンの炎症病態への影響
Oct. 2010

The role of ectopically-expressed adiponectin in rheumatoid arthritis.　　　　　　　　　　　　　　　
Haruyo Akiyama; Hiroya Shimada; Takaharu Negoro; Yasuko Nakano
14th International Congress of Immunology

非定型抗精神病薬の脂肪細胞分化に及ぼす影響の検討　　　　　　　　　　　　　　　
Mar. 2010

非定型抗精神病薬の脂肪細胞分化及びアディポネクチン分泌に及ぼす影響の検討　　　　　　　　　　　　　　　
Sep. 2009

LPSによる炎症誘導時のHMWアディポネクチンの機能　　　　　　　　　　　　　　　
Sep. 2009

末梢血幹細胞動員における線溶系因子の機能解析　　　　　　　　　　　　　　　
2009

c-kit欠損W/Wvマウスにおけるタンパク質の経口感作とその関連遺伝子　　　　　　　　　　　　　　　
2008

オボムコイ ド（OVM）のマウス経口感作について　　　　　　　　　　　　　　　
Oct. 2007

末梢血幹細胞動員における線溶系因子の機能解析　　　　　　　　　　　　　　　
Sep. 2007

血管病変形成過程における骨髄由来炎症性細胞動員の意義　　　　　　　　　　　　　　　
Sep. 2007

オボムコイド(OVM)のマウス経口感作について
Sep. 2007

線溶系因子活性化による骨髄ニッチ制御機構　　　　　　　　　　　　　　　
Sep. 2007

食物アレルゲン（オボムコイド）のマウス経 口感作への油脂の影響について　　　　　　　　　　　　　　　
Sep. 2007

The effect of plant oil on oral sensitization of mice with ovomucoid.　　　　　　　　　　　　　　　
Teshima R.; Okunuki H.; Nakamura R.; Sawada J.
13th International congress of mucosal immunology, Jul. 2007

造血幹細胞における線溶系因子の機能解析　　　　　　　　　　　　　　　
Mar. 2007

アテローム硬化性プラーク形成と炎症性細胞動員機構との関連　　　　　　　　　　　　　　　
Mar. 2007

オボムコイド(OVM)のマウス経口感作への油脂の影響について　　　　　　　　　　　　　　　
Mar. 2007

組換え食品中のCry1Abと食物アレルギー患者血清との反応性評価の研究　　　　　　　　　　　　　　　
Mar. 2007

アテローム硬化性プラーク形成と炎症性細胞動員機構との関連　　　　　　　　　　　　　　　
Oct. 2006

造血幹細胞における線溶系因子の機能解析　　　　　　　　　　　　　　　
Oct. 2006

骨髄Nicheにおける線溶系因子の機能解析　　　　　　　　　　　　　　　
Sep. 2006

W/Wv マウスの卵白アルブミン（OVA）経 口投与によるASA 誘導ならびにPAFの作用について　　　　　　　　　　　　　　　
Oct. 2005

一般小児血清中の免疫指標調査 IgE,TARCおよびIP-10濃度について
Sep. 2005

細胞の遊走の制御・ニッシェ 骨髄Nicheからの細胞動員と組織再生機構
Jul. 2005

Gene expression profiling of glucocorticoid-treated mast cells　　　　　　　　　　　　　　　
Nakamura, R.; Okunuki, H.; Ishida, S.; Ozawa, S.; Saito, Y.; Teshima, R.; Sawada, J.
The World Allergy Congress in Munich, Jun. 2005

BALB/cマウスにおけるCpG-ODN-OVA経鼻投与の効果　　　　　　　　　　　　　　　
Mar. 2005

3歳児血清中TARC濃度とアレルギー性疾患の関連性に関する検討　　　　　　　　　　　　　　　
Mar. 2005

マスト細胞におけるdibutylphthalateおよび抗原刺激によるEgr-1の発現誘導　　　　　　　　　　　　　　　
Mar. 2005

W/W^vマウスの卵白アルブミン(OVA)経口投与によるASA誘導ならびにPAFの作用について　　　　　　　　　　　　　　　
2005

W/WvマウスにおけるCpG ODN-OVA経口感作による影響について
Sep. 2004

3歳児の食物並びに吸入アレルゲン特異的IgE抗体の実態調査
Sep. 2004

3歳児の食物ならびに吸入アレルゲン特異的IgE抗体価の実態調査(第2報)
Mar. 2004

組換え食品中の新規蛋白質と患者血清の反応性評価法に関する研究　　　　　　　　　　　　　　　
Mar. 2004

W/WVマウスにおけるCpG Oligodeoxynucleotides-OVA経口感作による影響　　　　　　　　　　　　　　　
Mar. 2004

フタル酸エステル類のマスト細胞に及ぼすgenetic及びnon-geneticな影響　　　　　　　　　　　　　　　
Mar. 2004

放射性組換え抗体の脳内送達法の検討　　　　　　　　　　　　　　　
2004

3才児の食物並びに吸入アレルゲン特異的IgE抗体の実態調査　　　　　　　　　　　　　　　
2004

W/WVマウスにおけるOVA経口感作によるASA誘導とγδ-T細胞について
Sep. 2003

W/WvマウスのOvalbuminの経口感作とTCRγδ-T細胞における減少の関与の可能性(Oral sensitization of W/Wv mice with ovalbumin and possible involvement of the decrease in TCRγδ-T cells)　　　　　　　　　　　　　　　
Okunuki Haruyo; Teshima Reiko; Sato Yuji; Akiyama Hiroshi; Maitani Tamio; Sawada Jun-ichi
生化学, Aug. 2003, (公社)日本生化学会

鶏の卵白オボムコイドならびにそのPepsinフラグメントの消化剤安定性と潜在的アレルゲン性(Digestive stability and allergenic potential of chicken egg white ovomucoid and their pepsin-fragments)　　　　　　　　　　　　　　　
Takagi Kayoko; Teshima Reiko; Okunuki Haruyo; Itoh Satsuki; Kawasaki Nana; Kawanishi Toru; Hayakawa Takao; Sawada Jun-ichi
生化学, Aug. 2003, (公社)日本生化学会

遺伝子組換えとうもろこしCBH-351のラット並びにマウスへの90日間混餌投与による免疫系への影響　　　　　　　　　　　　　　　
Mar. 2003

マスト細胞の活性化に及ぼすステロイドホルモンの影響　　　　　　　　　　　　　　　
Mar. 2003

W/WvマウスにおけるOVA経口感作によるASA誘導とγδ-T細胞の減少　　　　　　　　　　　　　　　
Mar. 2003

3歳児の食物ならびに吸入アレルゲン特異的IgE抗体価の実態調査
Mar. 2003

食物抗原の人工胃腸液分解性に及ぼす加熱処理の影響　　　　　　　　　　　　　　　
Mar. 2003

W/WVマウスの卵白アルブミン(OVA)経口投与によるASA誘導について
Oct. 2002

食物アレルギー誘発性試験の一環として　　　　　　　　　　　　　　　
Sep. 2002

RBL-2H3細胞の遺伝子発現に及ぼすdexamethasoneの影響　　　　　　　　　　　　　　　
Aug. 2002

W/WVマウスにおけるOVA経口感作によるASA誘導　　　　　　　　　　　　　　　
Mar. 2002

遺伝子組換え食品導入蛋白質(CP4-EPSPS)の熱感受性並びにin vitro分解性試験　　　　　　　　　　　　　　　
Apr. 2001

転写レベルにおけるマスト細胞からのMCP-1産生制御機構について　　　　　　　　　　　　　　　
Aug. 2000

Examination of;active;systemic anaphylaxis in oral;immunized-mast cell deficient mice　　　　　　　　　　　　　　　
J. Sakushima; H. Okunuki; H. Akiyama; Y. Goda; M. Toyoda; R. Teshima; J. Sawada
18th International Congress of Biochemistry and Molecular Biology Beyond the Genome (Birmingham, UK)

Determination of enzymatic activity of EPSPS by radio-HPLC　　　　　　　　　　　　　　　
H. Okunuki, R. Teshima, H. Akiyama, Y. Goda, M. Toyoda, J. Sawada
18th International Congress of Biochemistry and Molecular Biology Beyond the Genome (Birmingham, UK)

遺伝子組換え,非組換え大豆のマウス,ラットへの混餌投与による免疫系への影響　　　　　　　　　　　　　　　
Mar. 2000

トルエンジイソシアネート曝露マウスにおける免疫影響指標の解析 (1) 液性免疫能(抗体産生)に関する指標　　　　　　　　　　　　　　　
2000

マスト細胞からのMCP-1産生機構について
Sep. 1999

HPLCによるシキミ酸経路関連酵素の活性測定法の検討　　　　　　　　　　　　　　　
Aug. 1999

培養マスト細胞からのカルシウム濃度上昇に伴う各種サイトカイン産生について　　　　　　　　　　　　　　　
1999

Functional analysis of adipocyte-specific Gelatin-binding protein of 28 kDa (GBP28) using diabetic mice　　　　　　　　　　　　　　　
M. Yoda; Y. Nakano; S. Enya; H. Okunuki; T. Tobe; M. Tomita
EIGHTH INTERNATIONAL CONGRESS ON OBESITY (Paris. France)

Genomic Structure and Promoter Analysis of the Gelatin-binding protein of 28 kDa (GBP28) gene　　　　　　　　　　　　　　　
S. Enya; K. Saito; T. Tobe; Y. Nakano; M. Yoda; H. Okunuki; S. Asakawa; S.Minoshima; N. Shimizu; M. Tomita
EIGHTH INTERNATIONAL CONGRESS ON OBESITY (Paris. France)

Functional analysis of the adipocyte-specific Gelatin-binding protein of 28 kDa (GBP28)　　　　　　　　　　　　　　　
H. Okunuki; Y. Nakano; M.Yoda; S. Enya; T. Tobe; M. Tomita
EIGHTH INTERNATIONAL CONGRESS ON OBESITY (Paris. France)

新規脂肪組織特異的蛋白質GBP28の生理機能に関する研究　　　　　　　　　　　　　　　
Aug. 1998

脂肪細胞特異的蛋白質GBP28の生理機能に関する研究　　　　　　　　　　　　　　　
Mar. 1998

IgE架橋を近接ライゲーションアッセイで捉える高感度セルフリーアレルギー試験の開発
Apr. 2023 - Mar. 2027

IgEの架橋活性を指標とする新規セルフリーアレルギー試験法の開発
01 Apr. 2020 - 31 Mar. 2023

Influence of environmental microplastic to IgE crosslinking ability of food allergens
Grant-in-Aid for Scientific Research (C)
Teikyo Heisei University
01 Apr. 2017 - 31 Mar. 2020
In recent years, many reports show that massive amount of microplastic (MP) floats on the surface of the global ocean. It was examined whether IgE crosslinking ability of allergens change by absorption and binding to MP.
The EXiLE method, which quantify IgE crosslinking on plasma membrane, was used in this study. Egg white allergen absorbed by 2.5 w/v% polystyrene beads whose diameter were 0.5μm or longer gave much higher luciferase activity than free allergen, demonstrating its IgE crosslinking ability was increased by binding to the beads. Further increase of those responses were observed when the reactions were carried out on a 96-well plate ("solid phase EXiLE"), indicating larger surface area bring increased response in the assay.

Study of the mechanism and therapeutic biomarkers for sublingual immunotherapy by using the EXiLE method
Grant-in-Aid for Scientific Research (B)
National Institute of Health Sciences
01 Apr. 2016 - 31 Mar. 2019
In this study we aimed to explore a biomarker that can predict the clinical response to sublingual immunotherapy (SLIT). Patients allergic to Japanese cedar pollen (JCP) were administered with JCP SLIT for more than 2 seasons. The severity of symptoms of pollinosis was represented by an averaged total nasal symptoms medication score (TNSMS) in the peak season. Crosslinking ability of patients’ serum IgE on the addition of JCP extract was determined by our original cell-based allergy test; IgE crosslinking-induced luciferase expression (EXiLE) method. When the reporter cells were sensitized with patients’ sera collected before starting SLIT, JCP-induced EXiLE response of more than 2.0 fold was an effective threshold to distinguish good and poor responders in SLIT judged by TNSMS diference (P<0.001). These results suggest that specific IgE crosslinking ability at SLIT entry would be a good biomarker candidate to predict the clinical response to SLIT.

アテローム硬化性プラーク形成と炎症性細胞動員機構との関連
2006 - 2007