Potter Timothy M, Skoczen Sarah L, Rodriguez Jamie C, Neun Barry W, Ilinskaya Anna N, Cedrone Edward, Dobrovolskaia Marina A
Cancer Research Technology Program, Nanotechnology Characterization Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, P.O. Box B, Frederick, MD, 21702, USA.
Methods Mol Biol. 2018;1682:125-133. doi: 10.1007/978-1-4939-7352-1_11.
This chapter provides a protocol for analysis of nanoparticle effects on the function of phagocytic cells. The protocol relies on luminol chemiluminescence to detect zymosan uptake. Zymosan is an yeast particle which is typically eliminated by phagocytic cells via the complement receptor pathway. The luminol, co-internalized with zymosan, is processed inside the phagosome to generate a chemiluminescent signal. If a test nanoparticle affects the phagocytic function of the cell, the amount of phagocytosed zymosan and, proportionally, the level of generated chemiluminescent signal change. Comparing the zymosan uptake of untreated cells with that of cells exposed to a nanoparticle provides information about the nanoparticle's effects on the normal phagocytic function. This method has been described previously and is presented herein with several changes. The revised method includes details about nanoparticle concentration selection, updated experimental procedure, and examples of the method performance.
本章提供了一种分析纳米颗粒对吞噬细胞功能影响的实验方案。该方案依靠鲁米诺化学发光来检测酵母聚糖的摄取。酵母聚糖是一种酵母颗粒，通常由吞噬细胞通过补体受体途径清除。与酵母聚糖共同内化的鲁米诺在吞噬体内被处理以产生化学发光信号。如果测试纳米颗粒影响细胞的吞噬功能，吞噬的酵母聚糖量以及相应产生的化学发光信号水平会发生变化。将未处理细胞与暴露于纳米颗粒的细胞的酵母聚糖摄取情况进行比较，可提供有关纳米颗粒对正常吞噬功能影响的信息。此方法先前已有描述，本文在此基础上做了一些修改。修订后的方法包括纳米颗粒浓度选择的详细信息、更新后的实验步骤以及方法性能示例。
