Use of bone marrow-derived macrophages to model murine innate immune responses
Use of bone marrow-derived macrophages to model murine innate immune responses
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DOI：
10.1016/j.jim.2006.01.017
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发表时间：
2006-04-20
影响因子：
2.2
通讯作者：
Heisterkamp, Nora
中科院分区：
文献类型：
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作者：
Cunnick, Jess;Kaur, Pavinder;Heisterkamp, Nora
先天免疫系统由中性粒细胞和单核细胞/巨噬细胞组成。作为一种细胞类型，骨髓源性巨噬细胞（BMM）比中性粒细胞更容易研究，因为它们仍然能够进行细胞分裂并且具有更长的寿命。然而，与中性粒细胞相比，很少有方法学研究的生产活性氧（ROS）的巨噬细胞已被报道。在这里，我们目前的研究在各种条件下，包括使用不同的引发剂和刺激剂的这种细胞类型的ROS生产。此外，我们报告说，从头粘附的BMM组织培养板诱导超氧阴离子的产生，这可以进一步增强PMA的刺激。BMM能够粘附于已被TNF-α激活的内皮细胞。暴露，并且在这些情况下也产生ROS。我们探索了不同的方法将基因产物引入BMM而不激活它们，以避免使随后的ROS产生研究复杂化。用慢病毒载体感染是非常有效的，允许长期表达并且不激活BMM。因此，BMM非常适合于氧化猝发的生化研究。(c)2006 Elsevier B. V.保留所有权利。
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.