Biodistribution of (89)Zr-DFO-labeled avian pathogenic Escherichia coli outer membrane vesicles by PET imaging in chickens.
Biodistribution of (89)Zr-DFO-labeled avian pathogenic Escherichia coli outer membrane vesicles by PET imaging in chickens.
复制标题
通过 PET 成像观察 (89)Zr-DFO 标记的禽致病性大肠杆菌外膜囊泡在鸡体内的生物分布。
DOI：
10.1016/j.psj.2022.102364
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发表时间：
2023-02
期刊：
影响因子：
4.4
通讯作者：
Jian Tu
中科院分区：
文献类型：
--
作者：
Zhe Li;Lulu Niu;Lizhen Wang;Ting Mei;Wenbin Shang;Xi Cheng;Yuqing Li;Feng Xi;Xiangjun Song;Ying Shao;Yuping Xu;Jian Tu
禽致病性大肠埃希菌（APEC）是家禽感染的一种严重的全身性传染病，给养禽业造成严重的经济损失。此前的研究表明，APEC通过分泌系统发挥致病性需要分泌毒力蛋白。外膜囊泡（OMV）是革兰氏阴性菌的普遍分泌系统，在毒力因子的长距离传递中发挥关键作用，但其是否与APEC的致病机制有关尚未确定。在本研究中，通过超速离心和密度梯度离心从 AE17（O2 血清型）中纯化和表征 OMV，并使用液相色谱-串联质谱 (LC-MS/MS) 鉴定其蛋白质货物。此外，用DFO螯合AE17 OMV后标记89Zr，并利用正电子发射断层扫描PET成像追踪鸡体内的89Zr-DFO-OMV并对其分布位点进行病理分析。这项研究表明，AE17 OMV 是大小范围为 20 至 200 nm 的膜囊泡，蛋白质组分析揭示了毒力蛋白的存在，包括粘附蛋白 OmpA、OmpC、OmpF、OmpX、FimH、FimC 和 FigE，以及血清抗性蛋白 OmpT 和 MliC 以及免疫反应调节蛋白 (FliC)。此外，追踪AE17 OMVs生物分布的体内PET成像显示，AE17 OMVs被肺部区域、胃肠道和肾脏区域吸收，但在其他区域没有检测到。对摄入 AE17 OMV 的组织部位进行病理分析，结果显示炎症反应和损伤。这些发现表明，AE17 OMV不仅含有一组与AE17感染相关的毒力蛋白，而且还可以长距离传递这些毒力蛋白并引起组织炎症损伤。我们的研究揭示了 APEC 发病机制中先前未识别的致病微生物信号，这可能有助于开发有效对抗 APEC 的疫苗和抗生素。
Avian pathogenic Escherichia coli (APEC) is a serious systemic infectious disease in poultry infections, causing severe economic losses to the poultry industry. Previous studies have shown that secretion of virulence proteins was required for the pathogenicity of APEC through the secretion system. Outer membrane vesicles (OMVs) are a generalized secretion system of Gram-negative bacteria that play a key role in the long-distance delivery of virulence factors, but whether they are associated with the pathogenic mechanism of APEC has not been determined. In this study, OMVs were purified and characterized from AE17 (O2 serotype) by ultracentrifugation and density gradient centrifugation and their protein cargo was identified using liquid chromatography-tandem mass spectrometry (LC-MS/MS). In addition, 89Zr was labeled after chelating AE17 OMVs by DFO and positron emission tomography PET imaging was used to track 89Zr-DFO-OMVs in chickens and to pathologically analyze the distribution sites. This study showed that AE17 OMVs were membrane vesicles ranging in size from 20 to 200 nm and proteomic analysis revealed the presence of virulence proteins, including adhesion proteins OmpA, OmpC, OmpF, OmpX, FimH, FimC and FigE, and serum resistance proteins OmpT and MliC and immune response regulator proteins (FliC). In addition, in vivo PET imaging to track the biodistribution of AE17 OMVs showed that AE17 OMVs were taken up by the lung region and the gastrointestinal and renal regions but were not detected in other areas. Pathological analysis of the tissue sites where AE17 OMVs were ingested showed inflammatory responses and damage. These findings suggested that AE17 OMVs not only contained a group of virulence proteins associated with AE17 infection but can also deliver these virulence proteins over long distances and caused tissue inflammatory damage. Our study revealed a previously unidentified causative microbial signal in the pathogenesis of APEC that could aid in the development of vaccines and antibiotics effective against APEC.