Korea Institute of Medical Microrobotics (KIMIRo), Gwangju 61011, Korea.
Laboratory Animal Center, Daegu-Gyeongbuk Medical Innovation Foundation, Daegu 700-721, Korea.
Int J Mol Sci. 2020 Aug 27;21(17):6205. doi: 10.3390/ijms21176205.
This study was conducted to monitor the macrophage infiltration of atopic dermatitis (AD)-like skin lesions and to evaluate the effects of anti-AD therapeutic agents in immunocompetent mice via optical reporter-gene-based molecular imaging. The enhanced firefly luciferase (effluc)-expressing macrophage cell line (Raw264.7/effluc) was intravenously introduced into mice with 2,4-dinitrochlorobenzene (DNCB)-induced AD, followed by bioluminescent imaging (BLI). After in vivo imaging, AD-like skin lesions were excised, and ex vivo imaging and Western blotting were conducted to determine the presence of infused macrophages. Finally, the therapeutic effect of dexamethasone (DEX), an AD-modulating agent, was evaluated via macrophage tracking. In vivo imaging with BLI revealed the migration of the reporter macrophages to DNCB-induced AD-like skin lesions on day 1 post-transfer. The greatest recruitment was observed on day 3, and a decline in BLI signal was observed on day 14. Notably, in vivo BLI clearly showed the inhibition of the reporter macrophage infiltration of DNCB-induced AD-like skin lesions by DEX, which was consistent with the reduced AD symptoms observed in DEX-treated mice. We successfully visualized the macrophage migration to DNCB-induced AD-like skin lesions, proving the feasibility of macrophage imaging for evaluating AD-regulating drugs in living organisms.
本研究旨在通过光学报告基因分子成像监测特应性皮炎(AD)样皮肤损伤中的巨噬细胞浸润,并评估免疫功能正常的小鼠中抗 AD 治疗药物的效果。将增强型萤火虫荧光素酶(effluc)表达的巨噬细胞系(Raw264.7/effluc)通过静脉注射到 2,4-二硝基氯苯(DNCB)诱导的 AD 小鼠中,然后进行生物发光成像(BLI)。在体内成像后,切除 AD 样皮肤损伤,并进行离体成像和 Western blot 分析,以确定输注的巨噬细胞的存在。最后,通过巨噬细胞追踪评估 AD 调节剂地塞米松(DEX)的治疗效果。BLI 的体内成像显示报告巨噬细胞在转移后第 1 天迁移到 DNCB 诱导的 AD 样皮肤损伤处。第 3 天观察到最大招募,第 14 天 BLI 信号下降。值得注意的是,体内 BLI 清楚地显示了 DEX 抑制 DNCB 诱导的 AD 样皮肤损伤中报告巨噬细胞的浸润,这与在 DEX 治疗的小鼠中观察到的 AD 症状减轻一致。我们成功地可视化了巨噬细胞向 DNCB 诱导的 AD 样皮肤损伤的迁移,证明了巨噬细胞成像用于评估活体 AD 调节药物的可行性。