Novel IgE crosslinking-induced luciferase expression method using human-rat chimeric IgE receptor-carrying mast cells.

Autor: Akiyama H; Division of Pharmacotherapeutics, Faculty of Pharmaceutical Sciences, Teikyo Heisei University, Tokyo, Japan. Electronic address: h.akiyama@thu.ac.jp., Kurisaka C; Division of Pharmacotherapeutics, Faculty of Pharmaceutical Sciences, Teikyo Heisei University, Tokyo, Japan., Kumasaka K; Chemistry Division, Kanagawa Prefectural Institute of Public Health, Kanagawa, Japan., Nakamura R; Division of Medicinal Safety Science, National Institute of Health Sciences, Kawasaki, Japan.
Jazyk: angličtina
Zdroj: Journal of immunological methods [J Immunol Methods] 2024 Jun; Vol. 529, pp. 113682. Date of Electronic Publication: 2024 May 04.
DOI: 10.1016/j.jim.2024.113682
Abstrakt: 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.
Competing Interests: Declaration of competing interest The authors declare no conflicts of interest.
(Copyright © 2024 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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