Upregulation of IL-4 receptor signaling pathway in circulating ILC2s from asthma patients

Rie Baba; Hiroki Kabata; Yoshitaka Shirasaki; Takashi Kamatani; Mai Yamagishi; Misato Irie; Risa Watanabe; Masako Matsusaka; Katsunori Masaki; Jun Miyata; Kazuyo Moro; Sotaro Uemura; Koichi Fukunaga

doi:10.1016/j.jacig.2022.07.007

Upregulation of IL-4 receptor signaling pathway in circulating ILC2s from asthma patients

J Allergy Clin Immunol Glob. 2022 Sep 12;1(4):299-304. doi: 10.1016/j.jacig.2022.07.007. eCollection 2022 Nov.

Authors

Rie Baba¹, Hiroki Kabata¹, Yoshitaka Shirasaki², Takashi Kamatani^{1

3

4

5}, Mai Yamagishi^{2

6}, Misato Irie¹, Risa Watanabe¹, Masako Matsusaka¹, Katsunori Masaki¹, Jun Miyata⁷, Kazuyo Moro^{8

9

10}, Sotaro Uemura¹¹, Koichi Fukunaga¹

Affiliations

¹ Division of Pulmonary Medicine, Department of Medicine, Keio University School of Medicine, Tokyo, Japan.
² Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
³ Laboratory for Medical Science Mathematics, Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan.
⁴ Department of AI Technology Development, M&D Data Science Center, Tokyo Medical and Dental University, Tokyo, Japan.
⁵ Division of Precision Cancer Medicine, Tokyo Medical and Dental University Hospital, Tokyo, Japan.
⁶ Live Cell Diagnosis, Ltd, Asaka, Saitama, Japan.
⁷ Division of Infectious Diseases and Respiratory Medicine, Department of Internal Medicine, National Defense Medical College, Saitama, Japan.
⁸ Laboratory for Innate Immune Systems, RIKEN Center for Integrative Medical Sciences, Yokohama, Kanagawa, Japan.
⁹ Laboratory for Innate Immune Systems, Department of Microbiology and Immunology, Graduate School of Medicine, Osaka University, Suita, Osaka, Japan.
¹⁰ Laboratory for Innate Immune Systems, Immunology Frontier Research Center, Osaka University, Suita, Osaka, Japan.
¹¹ Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan.

Abstract

Background: Group 2 innate lymphoid cells (ILC2s) produce type 2 cytokines by stimulation with epithelial cell-derived cytokines and are implicated in the pathogenesis of various allergic diseases, including asthma. However, differences in the molecular characteristics of ILC2s between patients with asthma and healthy subjects remain unclear.

Objective: We sought to evaluate differences in cytokine production capacity and gene expression profile of ILC2s in the peripheral blood of patients with asthma and healthy subjects.

Methods: We evaluated ILC2s derived from 15 patients with asthma and 7 healthy subjects using flow cytometry, live-cell imaging of secretion activity analysis, and RNA-sequencing.

Results: ILC2s were sorted as CD45⁺Lineage^-CRTH2⁺CD127⁺CD161⁺ cells from the peripheral blood of patients with asthma and healthy subjects, and the number of ILC2s was decreased in patients with asthma (851 ± 1134 vs 2679 ± 3009 cells/20 mL blood; P = .0066). However, patient-derived ILC2s were activated to produce more IL-5 and IL-13 in response to stimulation with IL-2, IL-33, and thymic stromal lymphopoietin compared with healthy subject-derived ILC2s (P = .0032 and P = .0085, respectively). Furthermore, RNA-sequencing analysis revealed that patient-derived ILC2s had different gene expression profiles, such as increased expression in cell growth-related genes (CDKN1b, CCNG2, CCND2, CCN1), prostaglandin E receptor (PTGER2), and IL-4 receptor. In addition, a gene set of the IL-4 receptor signaling pathway was significantly upregulated in ILC2s in patients with asthma (P = .042).

Conclusions: Our results suggest that circulating ILC2s in patients with asthma are preactivated via the IL-4 receptor signaling pathway and produce IL-5 and IL-13 vigorously by stimulation.

Keywords: Asthma; biomarker; group 2 innate lymphoid cells; live cell imaging.