Laboratory of Clinical Microbiology, Department of Microbiology, Immunology and Transplantation, Katholieke Universiteit (KU) Leuven, 3000 Leuven, Belgium.
Fungal Biology Group, School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, United Kingdom.
Dis Model Mech. 2022 Mar 1;15(3). doi: 10.1242/dmm.049165. Epub 2022 Mar 30.
Invasive pulmonary aspergillosis (IPA) caused by the mold Aspergillus fumigatus is one of the most important life-threatening infections in immunocompromised patients. The alarming increase of isolates resistant to the first-line recommended antifungal therapy urges more insights into triazole-resistant A. fumigatus infections. In this study, we systematically optimized a longitudinal multimodal imaging-compatible neutropenic mouse model of IPA. Reproducible rates of pulmonary infection were achieved through immunosuppression (sustained neutropenia) with 150 mg/kg cyclophosphamide at day -4, -1 and 2, and an orotracheal inoculation route in both sexes. Furthermore, increased sensitivity of in vivo bioluminescence imaging for fungal burden detection, as early as the day after infection, was achieved by optimizing luciferin dosing and through engineering isogenic red-shifted bioluminescent A. fumigatus strains, one wild type and two triazole-resistant mutants. We successfully tested appropriate and inappropriate antifungal treatment scenarios in vivo with our optimized multimodal imaging strategy, according to the in vitro susceptibility of our luminescent fungal strains. Therefore, we provide novel essential mouse models with sensitive imaging tools for investigating IPA development and therapy in triazole-susceptible and triazole-resistant scenarios.
由烟曲霉引起的侵袭性肺曲霉病（IPA）是免疫功能低下患者中最具威胁生命的感染之一。耐一线推荐抗真菌治疗的分离株数量的惊人增加，促使人们更深入地了解唑类耐药烟曲霉感染。在这项研究中，我们系统地优化了一种纵向多模态成像兼容的中性粒细胞减少症小鼠 IPA 模型。通过在第-4、-1 和 2 天用 150mg/kg 环磷酰胺进行免疫抑制（持续中性粒细胞减少），并通过两性的经口气管内接种途径，实现了可重复的肺部感染率。此外，通过优化荧光素剂量和工程同基因红移生物发光烟曲霉菌株（野生型和两种唑类耐药突变体），实现了真菌负荷检测的体内生物发光成像的灵敏度提高，早在感染后一天即可实现。我们根据我们发光真菌菌株的体外药敏情况，成功地在体内测试了我们优化的多模态成像策略的适当和不适当的抗真菌治疗方案。因此，我们提供了具有敏感成像工具的新型基本小鼠模型，用于研究唑类敏感和唑类耐药情况下的 IPA 发展和治疗。
