Inhibitory kappaB kinase 2 activates airway epithelial cells to stimulate bone marrow macrophages.
Inhibitory kappaB kinase 2 activates airway epithelial cells to stimulate bone marrow macrophages.
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DOI：
10.1165/rcmb.2006-0245oc
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发表时间：
2007-01
影响因子：
6.4
通讯作者：
B. Mathew;G. Park;H. Cao;A. Azim;Xuerong Wang;R. V. van Breemen;R. Sadikot;J. Christman
中科院分区：
文献类型：
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作者：
B. Mathew;G. Park;H. Cao;A. Azim;Xuerong Wang;R. V. van Breemen;R. Sadikot;J. Christman
目前还不清楚是巨噬细胞还是气道上皮细胞主要对感染和炎症刺激作出反应，并在气道内启动细胞间炎症相互作用。我们假设气道上皮细胞是对环境刺激激活巨噬细胞的主要应答者。为了研究气道上皮细胞在巨噬细胞激活中的单侧作用，我们建立了一个体外系统，将小鼠原代气管上皮细胞（MTEC）和原代骨髓源性巨噬细胞（BMDM）一起在Transwell培养板中孵育一小段时间。在与巨噬细胞孵育前，用表达构成型活性IKK2 （Ad-cIKK2）、ad - β - gal或PBS的腺病毒载体转染MTEC 48小时。通过检测巨噬细胞表面CD11b的表达、NF-kappaB的激活、吞噬活性和活性氧的产生、环氧化酶(COX)-2基因的表达和前列腺素的产生来确定巨噬细胞的活化。CD68免疫染色和扫描电镜证实巨噬细胞粘附于上皮层。转染Ad-cIKK2的MTEC细胞产生增加的IL-6、小鼠gro - α、tnf - α和前列腺素（PG）E2。BMDM暴露于转染了Ad-cIKK2的MTEC，导致CD11b表达增加，巨噬细胞对上皮细胞层的粘附增加。在转染Ad-cIKK2的MTEC孵育的BMDM中，NF-kappaB激活、COX-2基因表达和PGD2合成也增加。这些数据表明，气道上皮细胞可能在产生导致巨噬细胞活化的炎症信号中起主要作用。
It has not been resolved whether macrophages or airway epithelial cells primarily respond to infectious and inflammatory stimuli and initiate a cell-to-cell inflammatory interaction within the airways. We hypothesized that the airway epithelial cells are primary responders that activate macrophages in response to environmental stimuli. To investigate the unilateral contribution of airway epithelial cells in the activation of macrophages, we developed an in vitro system in which the primary mouse tracheal epithelial cells (MTEC) and primary bone marrow-derived macrophages (BMDM) were incubated together for a brief period of time in a Transwell culture plate. MTEC were transfected with adenoviral vectors that express a constitutively active form of IKK2 (Ad-cIKK2), Ad-beta-Gal, or PBS for 48 h before incubating with the macrophages. Macrophage activation was determined by measuring surface expression of CD11b, activation of NF-kappaB, phagocytic activity and production of reactive oxygen species, and cyclooxygenase (COX)-2 gene expression and production of prostaglandins. Macrophage adherence to epithelial layer was confirmed by CD68 immunostaining and scanning electron microscopy. MTEC cells transfected with Ad-cIKK2 produced increased amounts of IL-6, mouse GRO-alpha, TNF-alpha, and prostaglandin (PG)E2. Exposure of BMDM to MTEC, transfected with Ad-cIKK2, led to an increase in the CD11b expression and increased adherence of macrophages to the epithelial cell layer. NF-kappaB activation, COX-2 gene expression, and PGD2 synthesis were also increased in BMDM that were incubated with MTEC transfected with Ad-cIKK2. These data suggest that airway epithelial cells potentially play a primary role in generating inflammatory signals that result in activation of macrophages.