Department of Immunoregulation, Institute of Medical Science, Tokyo Medical University, Tokyo, Japan.
Department of Immunogenetics, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan; present address: Reiwa Health Sciences University Higashi-ku, Fukuoka, Japan
ALTEX. 2023;40(2):204-216. doi: 10.14573/altex.2111181. Epub 2022 Feb 24.
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.
尽管已经开发出几种预测化学物质致敏潜力的体外检测方法，但没有一种方法可以区分化学性呼吸道致敏剂和皮肤致敏剂。最近，我们建立了一个新的三维树突状细胞（DC）共培养系统，该系统由人气道上皮细胞系、源自人外周单核细胞的未成熟 DC 和人肺成纤维细胞系组成。在该共培养系统中，与皮肤致敏剂相比，呼吸道致敏剂在 DC 中的关键共刺激分子 OX40 配体（OX40L）的 mRNA 表达增强，这对于辅助性 T 细胞 2（Th2）细胞分化很重要。在此，我们通过向 DC 共培养系统中刺激的 DC 中添加外周同种异体幼稚 CD4+T 细胞，建立了一个新的两步 DC/T 细胞共培养系统。在这个 DC/T 细胞共培养系统中，模型呼吸道致敏剂，但不是皮肤致敏剂，增强了主要 Th2 标志物白细胞介素 4（IL-4）的 mRNA 表达。为了提高通用性，在 DC 共培养系统中代替外周单核细胞，使用了称为 CD14-ML 的单核细胞衍生增殖细胞。与皮肤致敏剂相比，呼吸道致敏剂诱导 CD14-ML 中 OX40L 的 mRNA 表达增强。当将这些细胞系应用于具有外周同种异体幼稚 CD4+T 细胞的 DC/T 细胞共培养系统时，呼吸道致敏剂但不是皮肤致敏剂增强了 IL-4 的 mRNA 表达。因此，通过 T 细胞中 IL-4 的差异 mRNA 上调，该 DC/T 细胞共培养系统可能有助于区分呼吸道和皮肤致敏剂。
