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猪肾上皮细胞志贺毒素（Stx）糖鞘脂受体的结构洞察以及使用新糖脂加标糖囊泡抑制Stx介导的细胞损伤

Structural Insights into Shiga Toxin (Stx) Glycosphingolipid Receptors of Porcine Renal Epithelial Cells and Inhibition of Stx-Mediated Cellular Injury Using Neoglycolipid-Spiked Glycovesicles.

作者信息

Detzner Johanna, Gloerfeld Caroline, Pohlentz Gottfried, Legros Nadine, Humpf Hans-Ulrich, Mellmann Alexander, Karch Helge, Müthing Johannes

机构信息

Institute for Hygiene, University of Münster, 48149 Münster, Germany.

Institute for Food Chemistry, University of Münster, 48149 Münster, Germany.

出版信息

Microorganisms. 2019 Nov 19;7(11):582. doi: 10.3390/microorganisms7110582.

DOI:10.3390/microorganisms7110582
PMID:31752441
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6920957/
Abstract

Shiga toxin (Stx) producing (STEC) cause the edema disease in pigs by releasing the swine-pathogenic Stx2e subtype as the key virulence factor. Stx2e targets endothelial cells of animal organs including the kidney harboring the Stx receptor glycosphingolipids (GSLs) globotriaosylceramide (Gb3Cer, Galα1-4Galβ1-4Glcβ1-1Cer) and globotetraosylceramide (Gb4Cer, GalNAcβ1-3Galα1-4Galβ1-4Glcβ1-1Cer). Since the involvement of renal epithelial cells in the edema disease is unknown, in this study, we analyzed the porcine kidney epithelial cell lines, LLC-PK1 and PK-15, regarding the presence of Stx-binding GSLs, their sensitivity towards Stx2e, and the inhibitory potential of Gb3- and Gb4-neoglycolipids, carrying phosphatidylethanolamine (PE) as the lipid anchor, towards Stx2e. Immunochemical and mass spectrometric analysis revealed various Gb3Cer and Gb4Cer lipoforms as the dominant Stx-binding GSLs in both LLC-PK1 and PK-15 cells. A dihexosylceramide with proposed Galα1-4Gal-sequence (GalCer) was detected in PK-15 cells, whereas LLC-PK1 cells lacked this compound. Both cell lines were susceptible towards Stx2e with LLC-PK1 representing an extremely Stx2e-sensitive cell line. Gb3-PE and Gb4-PE applied as glycovesicles significantly reduced the cytotoxic activity of Stx2e towards LLC-PK1 cells, whereas only Gb4-PE exhibited some protection against Stx2e for PK-15 cells. This is the first report identifying Stx2e receptors of porcine kidney epithelial cells and providing first data on their Stx2e-mediated damage suggesting possible involvement in the edema disease.

摘要

产志贺毒素（Stx）的大肠杆菌（STEC）通过释放猪致病性Stx2e亚型作为关键毒力因子，引发猪的水肿病。Stx2e作用于动物器官的内皮细胞，这些器官包括肾脏，肾脏中含有Stx受体糖鞘脂（GSLs），即球三糖神经酰胺（Gb3Cer，Galα1-4Galβ1-4Glcβ1-1Cer）和球四糖神经酰胺（Gb4Cer，GalNAcβ1-3Galα1-4Galβ1-4Glcβ1-1Cer）。由于肾上皮细胞在水肿病中的作用尚不清楚，在本研究中，我们分析了猪肾上皮细胞系LLC-PK1和PK-15中Stx结合GSLs的存在情况、它们对Stx2e的敏感性，以及携带磷脂酰乙醇胺（PE）作为脂质锚的Gb3-和Gb4-新糖脂对Stx2e的抑制潜力。免疫化学和质谱分析显示，在LLC-PK1和PK-15细胞中，各种Gb3Cer和Gb4Cer脂型是主要的Stx结合GSLs。在PK-15细胞中检测到一种具有推测的Galα1-4Gal序列的二己糖神经酰胺（GalCer），而LLC-PK1细胞缺乏这种化合物。两种细胞系对Stx2e均敏感，其中LLC-PK1是对Stx2e极其敏感的细胞系。作为糖囊泡应用的Gb3-PE和Gb4-PE显著降低了Stx2e对LLC-PK1细胞的细胞毒活性，而只有Gb4-PE对PK-15细胞表现出对Stx2e的一定保护作用。这是首次报道鉴定猪肾上皮细胞的Stx2e受体，并提供关于它们在Stx2e介导的损伤方面的初步数据，提示其可能参与水肿病。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/4abb2cbf5c4b/microorganisms-07-00582-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/ed5fb80b0315/microorganisms-07-00582-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/71d15e5c14a5/microorganisms-07-00582-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/13dda74a0db6/microorganisms-07-00582-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/53a529414d79/microorganisms-07-00582-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/16d17e2b940b/microorganisms-07-00582-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/890ab6a3c7de/microorganisms-07-00582-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/a0bf84116ea4/microorganisms-07-00582-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/f4c5dfdf74a6/microorganisms-07-00582-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/4abb2cbf5c4b/microorganisms-07-00582-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/ed5fb80b0315/microorganisms-07-00582-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/71d15e5c14a5/microorganisms-07-00582-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/13dda74a0db6/microorganisms-07-00582-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/53a529414d79/microorganisms-07-00582-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/16d17e2b940b/microorganisms-07-00582-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/890ab6a3c7de/microorganisms-07-00582-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/a0bf84116ea4/microorganisms-07-00582-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/f4c5dfdf74a6/microorganisms-07-00582-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f9/6920957/4abb2cbf5c4b/microorganisms-07-00582-g009.jpg

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