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标题：Application of Human FcεRI α-Chain-Transfected RBL-2H3 Cells for Estimation of Active Serum IgE
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作者：Kayoko Takagi ; Ryosuke Nakamura ; Reiko Teshima 等
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：2003
卷号：26
期号：2
页码：252-255
DOI：10.1248/bpb.26.252
出版社：The Pharmaceutical Society of Japan
摘要：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.
关键词：FcεRIα;basophile;transfectant;human serum;IgE;degranulation