Spatial proteomics revealed a CX3CL1-dependent crosstalk between the urothelium and relocated macrophages through IL-6 during an acute bacterial infection in the urinary bladder
Spatial proteomics revealed a CX3CL1-dependent crosstalk between the urothelium and relocated macrophages through IL-6 during an acute bacterial infection in the urinary bladder
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DOI：
10.1038/s41385-020-0269-7
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发表时间：
2020-02-28
影响因子：
8
通讯作者：
Engel, Daniel R.
中科院分区：
文献类型：
--
作者：
Bottek, Jenny;Soun, Camille;Engel, Daniel R.
膀胱的尿路上皮是第一道防线。然而，致尿性大肠埃希氏菌(UPEC)损害尿路上皮并引起急性细菌感染。在这里，我们展示了巨噬细胞和尿路上皮之间的串扰，刺激巨噬细胞迁移到尿路上皮。利用MALDI-MSI和LC-MS/MS的空间蛋白质组学，一种新的算法揭示了巨噬细胞的空间激活和迁移。空间蛋白质组分析揭示了Myo9b和F4/80在感染的尿路上皮细胞中的共表达，表明感染后巨噬细胞已经进入尿路上皮细胞。免疫荧光显微镜还显示，尿路内巨噬细胞吞噬UPEC并清除中性粒细胞。MALDI-MSI对空间蛋白质组的进一步分析表明，IL-6在尿路上皮细胞中有较强的表达，局部抑制该分子可减少巨噬细胞向尿路上皮细胞的迁移，从而加重感染。抑制IL-6后，尿路上皮细胞基质金属蛋白酶和趋化因子Cx(3)CL1的表达减少。因此，在CX(3)CR1(GFP/GFP)小鼠中，没有CX(3)CL1信号的情况下，巨噬细胞向尿路上皮细胞的迁移减少。总之，本研究通过IL-6诱导Cx(3)CL1的表达，描述了感染的尿路上皮细胞和巨噬细胞之间的串扰。这种串扰有助于巨噬细胞重新定位到尿路上皮细胞，并减少膀胱内的细菌负荷。
The urothelium of the urinary bladder represents the first line of defense. However, uropathogenic E. coli (UPEC) damage the urothelium and cause acute bacterial infection. Here, we demonstrate the crosstalk between macrophages and the urothelium stimulating macrophage migration into the urothelium. Using spatial proteomics by MALDI-MSI and LC-MS/MS, a novel algorithm revealed the spatial activation and migration of macrophages. Analysis of the spatial proteome unravelled the coexpression of Myo9b and F4/80 in the infected urothelium, indicating that macrophages have entered the urothelium upon infection. Immunofluorescence microscopy additionally indicated that intraurothelial macrophages phagocytosed UPEC and eliminated neutrophils. Further analysis of the spatial proteome by MALDI-MSI showed strong expression of IL-6 in the urothelium and local inhibition of this molecule reduced macrophage migration into the urothelium and aggravated the infection. After IL-6 inhibition, the expression of matrix metalloproteinases and chemokines, such as CX(3)CL1 was reduced in the urothelium. Accordingly, macrophage migration into the urothelium was diminished in the absence of CX(3)CL1 signaling in Cx(3)cr1(gfp/gfp) mice. Conclusively, this study describes the crosstalk between the infected urothelium and macrophages through IL-6-induced CX(3)CL1 expression. Such crosstalk facilitates the relocation of macrophages into the urothelium and reduces bacterial burden in the urinary bladder.