2023 Fiscal Year Final Research Report
Development of a novel coculture system using human monocyte cell line-derived immature dendritic cells and T cells for evaluation of allergic sensitizing potential
| Project/Area Number |
21K12256
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 63030:Chemical substance influence on environment-related
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| Research Institution | Tokyo Medical University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
善本 隆之 東京医科大学, 医学部, 教授 (80202406)
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| Project Period (FY) |
2021-04-01 – 2024-03-31
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| Keywords | 呼吸器感作性 / アレルギー感作性 / 動物実験代替法 / in vitro細胞共培養系 / ヒト単球細胞株 / Th2細胞 |
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| Outline of Final Research Achievements |
In the present study, we first established DC coculture system consisting of human airway epithelial cell line, immature dendritic cells (DCs) derived from peripheral blood monocytes, and fibroblast cell line, which were cultured in individual scaffolds and then overlaid. After stimulation with sensitizers, the scaffold containing DCs was moved, attached to the bottom of a new plate, and further stimulated with allogeneic CD4+ T cells. This two-step DC/T coculture system successfully discriminated respiratory sensitizers from skin sensitizers by preferential upregulation of IL-4 mRNA. We also generated human monocyte cell line CD14-ML cells and applied them to the DC coculture system and two-step DC/T coculture system. Resultant both systems successfully discriminated respiratory sensitizers from skin sensitizers by preferential upregulation of OX40L and IL-4 mRNA, respectively.
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| Free Research Field |
免疫毒性
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| Academic Significance and Societal Importance of the Research Achievements |
喘息などの呼吸器アレルギーは、重篤になる場合も多く、最悪はアナフィラキシーショックにより死に至る場合がある。ところが、化粧品業界などで動物実験が禁止される最中、この呼吸器感作性のリスクを事前に予測可能な動物実験代替法はなく、早急の開発が急務となっている。本研究では、汎用性改善のため、ヒト末梢血単球細胞株CD14-MLを作製し、ヒト末梢血単球の代わりにこの細胞をDC共培養系やDC/T共培養系に応用すると、ヒト末梢血単球の時と同様に、呼吸器感作性の識別が可能であることを見出した。今後、感作性化学物質の数を増やし、より簡単で誰にでもできる評価法にし、OECDのテストガイドライン化を目指していきたい。
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