Open AccessA novel coculture system for assessing respiratory sensitizing potential by IL-4 in T cells
Author(s) -
Itaru Mizoguchi,
Yasuhiro Katahira,
Shinya Inoué,
Eri Sakamoto,
Aruma Watanabe,
Yuma Furusaka,
Atsushi Irie,
Satoru Senju,
Yasuharu Nishimura,
Shusaku Mizukami,
Kenji Hirayama,
Sou Nakamura,
Koji Eto,
Hideaki Hasegawa,
Takayuki Yoshimoto
Publication year - 2022
Publication title -
altex/alternatives to animal experimentation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.975
H-Index - 51
eISSN - 1868-8551
pISSN - 1868-596X
DOI - 10.14573/altex.2111181
Subject(s) - cd14 , dendritic cell , cell culture , fibroblast , immunology , monocyte , t cell , respiratory system , jurkat cells , cell , microbiology and biotechnology , biology , chemistry , immune system , biochemistry , anatomy , genetics
Although several in vitro assays that predict the sensitizing potential of chemicals have been developed, none can distinguish between chemical respiratory and skin sensitizers. Recently, we established a new three-dimensional dendritic cell (DC) coculture system consisting of a human airway epithelial cell line, immature DCs derived from human peripheral monocytes, and a human lung fibroblast cell line. In this coculture system, compared to skin sensitizers, respiratory sensitizers showed enhanced mRNA expression in DCs of the key costimulatory molecule OX40 ligand (OX40L), which is important for T helper 2 (Th2) cell differentiation. Herein, we established a new two-step DC/T cell coculture system by adding peripheral allogeneic naïve CD4+ T cells to the DCs stimulated in the DC coculture system. In this DC/T cell coculture system, model respiratory sensitizers, but not skin sensitizers, enhanced mRNA expression of the predominant Th2 marker interleukin-4 (IL-4). To improve the versatility, in place of peripheral monocytes, monocyte-derived proliferating cells called CD14-ML were used in the DC coculture system. As in peripheral monocytes, enhanced mRNA expression of OX40L was induced in CD14-ML by respiratory sensitizers compared to skin sensitizers. When these cell lines were applied to the DC/T cell coculture system with peripheral allogeneic naïve CD4+ T cells, respiratory sensitizers but not skin sensitizers enhanced the mRNA expression of IL-4. Thus, this DC/T cell coculture system may be useful for discriminating between respiratory and skin sensitizers by differential mRNA upregulation of IL-4 in T cells.