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

Haruyo Akiyama; Chisato Kurisaka; Kenichi Kumasaka; Ryosuke Nakamura

doi:10.1016/j.jim.2024.113682

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

J Immunol Methods. 2024 May 4:529:113682. doi: 10.1016/j.jim.2024.113682. Online ahead of print.

Authors

Haruyo Akiyama¹, Chisato Kurisaka², Kenichi Kumasaka³, Ryosuke Nakamura⁴

Affiliations

¹ Division of Pharmacotherapeutics, Faculty of Pharmaceutical Sciences, Teikyo Heisei University, Tokyo, Japan. Electronic address: h.akiyama@thu.ac.jp.
² Division of Pharmacotherapeutics, Faculty of Pharmaceutical Sciences, Teikyo Heisei University, Tokyo, Japan.
³ Chemistry Division, Kanagawa Prefectural Institute of Public Health, Kanagawa, Japan.
⁴ Division of Medicinal Safety Science, National Institute of Health Sciences, Kawasaki, Japan.

PMID: 38705372
DOI: 10.1016/j.jim.2024.113682

Abstract

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.

Keywords: Allergen-specific IgE; Allergy diagnosis; High-affinity IgE receptor; Luciferase; Mast cells.