Hope William W, Kruhlak Michael J, Lyman Caron A, Petraitiene Ruta, Petraitis Vidmantas, Francesconi Andrea, Kasai Miki, Mickiene Diana, Sein Tin, Peter Joanne, Kelaher Amy M, Hughes Johanna E, Cotton Margaret P, Cotten Catherine J, Bacher John, Tripathi Sanjay, Bermudez Louis, Maugel Timothy K, Zerfas Patricia M, Wingard John R, Drusano George L, Walsh Thomas J
Immunocompromised Host Section, Pediatric Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
J Infect Dis. 2007 Feb 1;195(3):455-66. doi: 10.1086/510535. Epub 2006 Dec 21.
Little is known about the pathogenesis of invasive pulmonary aspergillosis and the relationship between the kinetics of diagnostic markers and the outcome of antifungal therapy.
An in vitro model of the human alveolus, consisting of a bilayer of human alveolar epithelial and endothelial cells, was developed. An Aspergillus fumigatus strain expressing green fluorescent protein was used. Invasion of the cell bilayer was studied using confocal and electron microscopy. The kinetics of culture, polymerase chain reaction, and galactomannan were determined. Galactomannan was used to measure the antifungal effect of macrophages and amphotericin B. A mathematical model was developed, and results were bridged to humans.
A. fumigatus penetrated the cellular bilayer 14-16 h after inoculation. Galactomannan levels were inextricably tied to fungal invasion and were a robust measure of the antifungal effect of macrophages and amphotericin B. Neither amphotericin nor macrophages alone was able to suppress the growth of A. fumigatus; rather, the combination was required. Monte Carlo simulations showed that human dosages of amphotericin B of at least 0.6 mg/kg were required to achieve adequate drug exposure.
This model provides a strategy by which relationships among pathogenesis, immunological effectors, and antifungal drug therapy for invasive pulmonary aspergillosis may be further understood.
侵袭性肺曲霉病的发病机制以及诊断标志物动力学与抗真菌治疗结果之间的关系鲜为人知。
建立了一种由人肺泡上皮细胞和内皮细胞双层组成的人肺泡体外模型。使用表达绿色荧光蛋白的烟曲霉菌株。利用共聚焦显微镜和电子显微镜研究细胞双层的侵袭情况。测定培养、聚合酶链反应和半乳甘露聚糖的动力学。使用半乳甘露聚糖来测量巨噬细胞和两性霉素B的抗真菌作用。建立了一个数学模型，并将结果与人类情况相关联。
接种后14 - 16小时烟曲霉穿透细胞双层。半乳甘露聚糖水平与真菌侵袭密切相关，是巨噬细胞和两性霉素B抗真菌作用的有力指标。单独使用两性霉素或巨噬细胞都不能抑制烟曲霉的生长；相反，两者联合使用是必需的。蒙特卡罗模拟表明，人类使用两性霉素B的剂量至少为0.6mg/kg才能实现足够的药物暴露。
该模型提供了一种策略，通过此策略可进一步了解侵袭性肺曲霉病的发病机制、免疫效应器和抗真菌药物治疗之间的关系。
