Role of TNF receptor-associated factor 3 in the CD40 signaling by production of reactive oxygen species through association with p40phox, a cytosolic subunit of nicotinamide adenine dinucleotide phosphate oxidase
Role of TNF receptor-associated factor 3 in the CD40 signaling by production of reactive oxygen species through association with p40phox, a cytosolic subunit of nicotinamide adenine dinucleotide phosphate oxidase
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DOI：
10.4049/jimmunol.172.1.231
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发表时间：
2004-01-01
影响因子：
4.4
通讯作者：
Lee, JR
中科院分区：
文献类型：
--
作者：
Ha, YJ;Lee, JR
为了扩展我们以前的报告，其中显示的活性氧（ROS）的生产后，在B细胞中的CD 40连接，我们进一步研究了可能的机制，ROS的生产和参与CD 40诱导的ROS中的p38激活。我们的研究表明，用抗CD 40刺激WEHI 231 B淋巴瘤诱导ROS产生和p38活化。抗氧化剂N-乙酰-L-半胱氨酸或NADPH氧化酶抑制剂阻断了这两种作用，但5-脂氧合酶抑制剂没有。我们还表明，用磷脂酰肌醇3-激酶（PI 3-K）抑制剂处理细胞会干扰CD 40诱导的ROS产生和p38激活。此外，当用Rac 1（N17 Rac 1）或TNFR相关因子（TRAF）3的显性阴性形式过表达时，WEHI 231 B细胞没有显示出对CD 40刺激产生ROS的完全应答。分子缔合研究进一步揭示了TRAF 3与NADPH氧化酶的胞浆亚基p40（phox）和PI 3-K的亚基p85的缔合可能是WEHI 231 B细胞中由CD 40刺激产生ROS的原因。总的来说，这些数据表明，CD 40诱导的WEHI 231中NADPH氧化酶的ROS产生需要TRAF 3的作用，以及PI 3-K和Rac 1的活性。
To extend our previous report, which showed the production of the reactive oxygen species (ROS) after the CD40 ligation in the B cells, we further examined the possible mechanisms for ROS production and the involvement of CD40-induced ROS in p38 activation. Our research shows that the stimulation of WEHI 231 B lymphomas with anti-CD40 induced ROS production and p38 activation. An antioxidant N-acetyl-L-cysteine or an inhibitor for NADPH oxidase blocked both of these, but the inhibitors for 5-lipoxygenase did not. We also show that the treatment of cells with inhibitors for the phosphatidylinositol 3-kinase (PI3-K) interfered with the CD40-induced ROS production and p38 activation. In addition, when overexpressed with a dominant negative form of either Rac1 (N17Rac1) or the TNFR-associated factor (TRAF) 3, the WEHI 231 B cells did not show a full response to the CD40 stimulation to produce ROS. Molecular association studies further revealed that the TRAF3 association with p40(phox), a cytosolic subunit of NADPH oxidase and p85 (a subunit of PI3-K), may possibly be responsible for the production of ROS by CD40 stimulation in WEHI 231 B cells. Collectively, these data suggest that the CD40-induced ROS production by NADPH oxidase in WEHI 231 requires the role of TRAF3, as well as activities of PI3-K and Rac1.