Secreted MbovP0145 Promotes IL-8 Expression through Its Interactive β-Actin and MAPK Activation and Contributes to Neutrophil Migration.
Secreted MbovP0145 Promotes IL-8 Expression through Its Interactive β-Actin and MAPK Activation and Contributes to Neutrophil Migration.
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分泌的MbovP 0145通过其相互作用的β-肌动蛋白和MAPK活化促进IL-8表达并促进神经元迁移。
DOI：
10.3390/pathogens10121628
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发表时间：
2021-12-15
期刊：
影响因子：
--
通讯作者：
Guo A
中科院分区：
文献类型：
--
作者：
Lu D;Zhang H;Zhang Y;Zhao G;Anwar Khan F;Chen Y;Hu C;Yang L;Chen H;Guo A
牛支原体(Mycodium bovis，M.bovis)是一种重要的牛病原，在世界范围内给奶牛业和肉牛业造成了巨大的经济损失。牛分枝杆菌分泌的蛋白质主要与其黏附、侵袭、毒力和细胞内生存有关，并在支原体与宿主的相互作用中发挥作用。在我们之前的研究中，我们发现了MbovP0145，一种存在于牛分枝杆菌分泌组中的分泌性蛋白，但对其功能知之甚少。在这项研究中，我们评估了重组MbovP0145(RMbovP0145)的炎症特征和炎症机制。为此，用rMbovP0145刺激牛肺上皮细胞(EBL)以时间和剂量依赖的方式观察IL-8的产生。我们观察到rMbovP0145通过激活ERK1/2和P38途径增加IL-8的产生。此外，还证实了牛分枝杆菌ΔMBOV0145突变株及其互补株对IL-8mRNA表达的影响。GST标记的MbovP0145蛋白的质谱分析表明，β-肌动蛋白可以与rMbovP0145特异性地相互作用，介导IL-8信号转导。当β-肌动蛋白在EBL细胞中的表达被RNA干扰抑制时，IL-8mRNA的表达和磷酸化的ERK1/2和P38蛋白减少，而细胞松弛素D破坏肌动蛋白聚合导致IL-8的表达和MAPK的磷酸化显著增加。与牛分枝杆菌HB0801及其互补株相比，感染牛分枝杆菌ΔMbov_0145突变体的EBL细胞培养上清液诱导的中性粒细胞向Transwell系统下部的迁移较少。总之，MbovP0145通过激活β-肌动蛋白途径与MAPK-肌动蛋白相互作用，促进IL-8的表达，从而促进中性粒细胞的迁移。
Mycoplasma bovis (M. bovis) is an important pathogen of cattle responsible for huge economic losses in the dairy and beef industries worldwide. The proteins secreted by M. bovis are mainly related to its adhesion, invasion, virulence, and intracellular survival and play a role in mycoplasma–host interactions. In our previous study, we found MbovP0145, a secreted protein present in the M. bovis secretome, but little is known about its function. In this study, we assessed the inflammatory characteristics and underlined mechanism of this inflammation of recombinant MbovP0145 (rMbovP0145). For this, bovine lung epithelial cells (EBL) were stimulated by rMbovP0145 to see the IL-8 production in a time- and dose-dependent manner. We observed that rMbovP0145 increased the production of IL-8 via ERK1/2 and P38 pathway activation. Further, the effect of the M. bovis ΔMbov_0145 mutant and its complementary strain on IL-8 mRNA expression was also confirmed. A pulldown assay of the GST-tagged MbovP0145 protein with mass spectrometry demonstrated that β-actin could specifically interact with rMbovP0145 to mediate the IL-8 signaling. As knockdown of β-actin expression with RNA interference in EBL cells decreased the mRNA expression of IL-8 and the phosphorylated ERK1/2 and P38 proteins, whereas disrupted actin polymerization by cytochalasin D led to a significantly higher IL-8 expression and MAPK phosphorylation in rMbovP0145-stimulated cells. Compared to M. bovis HB0801 and its complementary strain, the culture supernatant of EBL cells infected with the M. bovis ΔMbov_0145 mutant induced less neutrophil migration to the lower chamber in a transwell system. In conclusion, MbovP0145 promoted IL-8 expression by interacting with β-actin through activation of the MAPK pathway, thus contributing to neutrophil migration.
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