Application of Human FcεRI α-Chain-Transfected RBL-2H3 Cells for Estimation of Active Serum IgE
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概要
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We transfected the α-chain of human FcεRI into rat basophilic leukemia cell line RBL-2H3, established several stable transfected cells, and screened them by β-hexosaminidase release induced by sensitization with human IgE and stimulation with anti-human IgE antibody. A cloned cell line RBL-hEIa-2B12 was the strongest responder among the transfected cell clones. The concentrations of cytosolic free Ca^2+ concentration in the human IgE-sensitized cells increased after stimulation with anti-human IgE antibody. Thus, it is suggested that the α-chain of human FcεRI is associated with the β-chain and/or γ-chain of rat FcεRI, and that they form functional high affinity IgE receptor complexes. The total IgE concentrations of the sera from allergic patients were determined by using the β-hexosaminidase release assay, where the transfected cells were sensitized with diluted and heat-inactivated (at 56℃ for 30 min) serum and stimulated with anti-human IgE antibody. The IgE concentration obtained correlated with those measured by an enzyme immunoassay method. β-Hexosaminidase release induced by stimulation with 5 times diluted serum was sometimes less than the release induced by the same serum; diluted 25 times or 125 times, suggesting that these serum contained factors that blocked IgE binding to FcεRI or cross-linking by anti-human IgE antibody. The results suggested that our system will be useful for detecting FcεRIα-bindable IgE in human serum.
- 公益社団法人日本薬学会の論文
- 2003-02-01
著者
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TESHIMA Reiko
National Institute of Health Sciences
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Sawada Junichi
Project Team For Pharmacogenetics National Institute Of Health Sciences
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Sawada Jun-ichi
National Institute Of Health Sciences
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Teshima Reiko
Division of Biochemistry and Immunochemistry, National Institute of Health Sciences
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Takagi Kayoko
Division Of Biochemistry And Immunochemistry National Institute Of Health Sciences (nihs)
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Takagi Kayoko
国立医薬品食品衛生研究所 機能生化学部
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Takagi Kayoko
Division Of Biochemistry And Immunochemistry National Institute Of Health Sciences
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Teshima R
National Inst. Of Health Sciences
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Teshima Reiko
国立医薬品食品衛生研究所 機能生化学部
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Teshima Reiko
名古屋市立大学 薬学研究科
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Nakamura R
Division Of Novel Foods And Immunochemistry National Institute Of Health Sciences (nihs)
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Teshima Reiko
National Institute Of Health Science
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TAKAGI Kayoko
National Institute of Health Sciences
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NAKAMURA Ryosuke
National Institute of Health Sciences, Division of Biochemistry and Immunochemistry
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Teshima Reiko
National Inst. Of Health Sciences
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Sawada J
Mie Univ. School Of Medicine Mie Jpn
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Takagi K
National Inst. Of Health Sci. Tokyo Jpn
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Sawada J
National Inst. Of Health Sci. Tokyo Jpn
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Nakamura Ryosuke
National Institute Of Advanced Industrial Science And Technology (aist)
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Nakamura Ryosuke
National Institute Of Advanced Industrial Science And Technology
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Nakamura Ryosuke
Division of Novel Foods and Immunochemistry, National Institute of Health Sciences (NIHS)
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