Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
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Application of Human FcεRI α-Chain-Transfected RBL-2H3 Cells for Estimation of Active Serum IgE
Kayoko TakagiRyosuke NakamuraReiko TeshimaJun-ichi Sawada
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2003 Volume 26 Issue 2 Pages 252-255

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Abstract

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 Ca2+ 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 °C 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.

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© 2003 The Pharmaceutical Society of Japan
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