Use of bone marrow-derived macrophages to model murine innate immune responses

被引:24
作者
Cunnick, Jess
Kaur, Pavinder
Cho, YoungJin
Groffen, John
Heisterkamp, Nora
机构
[1] Univ So Calif, Childrens Hosp, Div Hematol Oncol, Sect Mol Carcinogenesis,Saban Res Inst, Los Angeles, CA 90027 USA
[2] Univ So Calif, Keck Sch Med, Los Angeles, CA 90027 USA
关键词
bone marrow-derived macrophage; reactive oxygen species; lipopolysaccharide; endothelial cell;
D O I
10.1016/j.jim.2006.01.017
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The innate immune system is composed of neutrophils and monocyte/macrophages. As a cell type, bone marrow-derived macrophage (BMM) are easier to study than neutrophils since they are still capable of cell division and have a longer life span. However, in comparison with neutrophils, few methodological studies on the production of reactive oxygen species (ROS) by such macrophages have been reported. Here we present studies on ROS production of this cell type under various conditions including the use of different priming and stimulating agents. In addition, we report that the de novo adhesion of BMM to tissue culture plates induces superoxide anion production and this can be further enhanced by stimulation with PMA. BMM are able to adhere to endothelial cells that have been activated by TNF-alpha. exposure, and under these circumstances also generate ROS. We explored different methods to introduce gene products into BMM without activating them to avoid complicating subsequent studies of ROS production. Infection with lentiviral vectors was very efficient, allowed long-term expression and did not activate the BMM. We conclude that BMM are very suitable for the biochemical study of the oxidative burst. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:96 / 105
页数:10
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