Development of a novel alternative to animal testing for the evaluation of respiratory allergic sensitizing potential of chemicals

2020 Fiscal Year Final Research Report

• Project/Area Number: 18K11658
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 63030:Chemical substance influence on environment-related
• Research Institution: Tokyo Medical University
• Principal Investigator: Mizoguchi Izuru 東京医科大学, 医学部, 講師 (00569527)
• Co-Investigator(Kenkyū-buntansha): 善本 隆之 東京医科大学, 医学部, 教授 (80202406)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: 感作性評価法 / 動物実験代替法 / 呼吸器感作性 / 皮膚感作性 / Th2反応

Outline of Final Research Achievements

Recently, we established a new 3-dimentional (3D) dendritic cell (DC) coculture system consisting of human airway epithelial cell line, immature DCs derived from human peripheral monocytes, and lung fibroblast cell line. This coculture system was shown to successfully discriminate respiratory sensitizers from skin sensitizers using 6 representative chemical sensitizers by more enhanced mRNA expression of key costimulatory molecule OX40 ligand, which is important for Th2 differentiation, in DCs. Herein, we have further established a new 2-step 3D DC/T coculture system by introducing T cells in the DC coculture system, in which IL-4 expression in T cells, that are the Key event 4 in the adverse outcome pathway of sensitization, can be used as a marker to discriminate these sensitizers. To increase the versatility, we are also trying to generate DC progenitor cell lines and T cell lines to apply for it.

Free Research Field

免疫毒性

Academic Significance and Societal Importance of the Research Achievements

化学物質の感作性試験法には、近年、動物を使わない代替法の開発が急務となっている。感作性化学物質には、呼吸器と皮膚感作性の2種類あり、両者に対して講ずるべき危機管理対策のレベルが全く異なるにも関わらず、既存のin vitro感作性評価法では、両者を見分けることができない。また、これまでに、感作性の有害性発現経路のKey event 4のT細胞を指標にした代替試験法は開発されていない。本研究により、T細胞でのIL-4発現を指標に呼吸器と皮膚感作性を識別する樹状細胞とT細胞の3次元共培養系の開発の可能性が示唆され、さらに汎用性の高い系ができれば、産業界への影響は多大でその社会的意義も大きい。