FLUORESCENT LIPOSOMES AS QUANTITATIVE MARKERS OF PHAGOCYTOSIS BY ALVEOLAR MACROPHAGES
FLUORESCENT LIPOSOMES AS QUANTITATIVE MARKERS OF PHAGOCYTOSIS BY ALVEOLAR MACROPHAGES
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DOI：
10.1016/0022-1759(95)00011-x
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发表时间：
1995-04-26
影响因子：
2.2
通讯作者：
MARTIN, WJ
中科院分区：
文献类型：
--
作者：
PERRY, DG;MARTIN, WJ
本文介绍了一种基于肺泡巨噬细胞（AM）摄取脂质体的新的吞噬试验。将荧光微球封装在脂质体中，这允许通过荧光测定法进行快速计数。在玻连蛋白存在下制备的脂质体具有暴露在其外表面上的蛋白质，如通过免疫标记所确定的。在有利于吞噬作用的条件下孵育脂质体和大鼠AM。光学和电子显微镜观察显示AM吞噬脂质体，荧光标记从脂质体逐渐转移到细胞内部。这种转移是由于真正的吞噬作用，如（1）脂质体包封的二氢荧光素（DHF）-酵母聚糖仅在AM内的荧光和（2）酵母聚糖相关脂质体仅在允许吞噬作用的条件下触发呼吸爆发所证明的。吞噬活性表示为脂质体/细胞，即每个巨噬细胞吞噬的脂质体的平均数量。我们用这种技术来跟踪吞噬作用随时间的推移，并测量脂多糖（LPS）和玻连蛋白对AM吞噬作用的影响。
A new phagocytic assay based on liposome ingestion by alveolar macrophages (AMs) is described. Fluorescent microspheres were encapsulated in liposomes, which allowed rapid enumeration by fluorometry. Liposomes made in the presence of vitronectin had the protein exposed on their outer surfaces, as determined by immunolabelling. Liposomes and rat AMs were incubated under conditions favorable for phagocytosis. Observation by light and electron microscopy showed AMs engulfing liposomes, with gradual transfer of fluorescent label from liposome to cell interior. This transfer was due to bona fide phagocytosis, as evidenced by (1) fluorescence of liposome-encapsulated dihydrofluorescein (DHF)-zymosan exclusively within AMs and (2) triggering of the respiratory burst by zymosan-associated liposomes only under conditions that allowed phagocytosis. Phagocytic activity was expressed as liposomes/cell, the average number of liposomes phagocytosed per macrophage. We used this technique to follow phagocytosis over time and to measure the effects of lipopolysaccharide (LPS) and vitronectin on AM phagocytosis.