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首页> 外文期刊>Infection and immunity >Differential Effects of Escherichia coli Subtilase Cytotoxin and Shiga Toxin 2 on Chemokine and Proinflammatory Cytokine Expression in Human Macrophage, Colonic Epithelial, and Brain Microvascular Endothelial Cell Lines
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Differential Effects of Escherichia coli Subtilase Cytotoxin and Shiga Toxin 2 on Chemokine and Proinflammatory Cytokine Expression in Human Macrophage, Colonic Epithelial, and Brain Microvascular Endothelial Cell Lines

机译：大肠杆菌枯草杆菌蛋白酶细胞毒素和志贺毒素2对人巨噬细胞，结肠上皮和脑微血管内皮细胞系中趋化因子和促炎细胞因子表达的差异作用

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Subtilase cytotoxin (SubAB) is the prototype of a recently emerged family of AB5 cytotoxins produced by Shiga-toxigenic Escherichia coli (STEC). Its mechanism of action involves highly specific A-subunit-mediated proteolytic cleavage of the essential endoplasmic reticulum (ER) chaperone BiP. Our previous in vivo studies showed that intraperitoneal injection of purified SubAB causes a major redistribution of leukocytes and elevated leukocyte apoptosis in mice, as well as profound splenic atrophy. In the current study, we investigated selected chemokine and proinflammatory cytokine responses to treatment with SubAB, a nontoxic derivative (SubAA272B), or Shiga toxin 2 (Stx2) in human macrophage (U937), brain microvascular endothelial (HBMEC), and colonic epithelial (HCT-8) cell lines, at the levels of secreted protein, cell-associated protein, and gene expression. Stx2 treatment upregulated expression of chemokines and cytokines at both the protein and mRNA levels. In contrast, SubAB induced significant decreases in secreted interleukin-8 (IL-8) and monocyte chemoattractant protein 1 (MCP-1) in all three tested cell lines and a significant decrease in secreted IL-6 in HBMECs. The downregulation of secreted chemokines or cytokines was not observed in SubAA272B-treated cells, indicating a requirement for BiP cleavage. The downregulation of secreted chemokines and cytokines by SubAB was not reflected at the mRNA and cell-associated protein levels, suggesting a SubAB-induced export defect.
机译：枯草杆菌细胞毒素（SubAB）是由志贺毒原性大肠杆菌（STEC）产生的AB5细胞毒素家族的最近出现的原型。它的作用机制涉及基本内质网（ER）伴侣蛋白BiP的高特异性A亚基介导的蛋白水解裂解。我们先前的体内研究表明，腹腔内注射纯化的SubAB会引起小鼠白细胞的大量重新分布和白细胞凋亡的增加，以及严重的脾萎缩。在本研究中，我们研究了对人巨噬细胞（U937），脑微血管中的SubAB，无毒衍生物（SubA A272 B）或志贺毒素2（Stx2）的治疗所选择的趋化因子和促炎细胞因子的反应。内皮细胞（HBMEC）和结肠上皮细胞（HCT-8）的分泌蛋白，细胞相关蛋白和基因表达水平。 Stx2处理在蛋白质和mRNA水平上调了趋化因子和细胞因子的表达。相比之下，SubAB诱导所有三种测试细胞系中分泌的白介素8（IL-8）和单核细胞趋化蛋白1（MCP-1）显着降低，而HBMECs分泌的IL-6显着降低。在SubA A272 B处理的细胞中未观察到分泌的趋化因子或细胞因子的下调，表明需要BiP裂解。 SubAB对分泌的趋化因子和细胞因子的下调未反映在mRNA和与细胞相关的蛋白质水平上，这表明SubAB诱导的出口缺陷。

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