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首页> 外文期刊>Infection and immunity >Role of the Type III Secreted Exoenzymes S, T, and Y in Systemic Spread of Pseudomonas aeruginosa PAO1 In Vivo
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Role of the Type III Secreted Exoenzymes S, T, and Y in Systemic Spread of Pseudomonas aeruginosa PAO1 In Vivo

机译：III型分泌的外切酶S，T和Y在铜绿假单胞菌PAO1体内系统传播中的作用

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Pseudomonas aeruginosa uses a dedicated type III secretion system to deliver toxins directly into the cytoplasm of host cells. While progress has been made in elucidating the function of type III-secreted toxins in vitro, the in vivo functions of the type III-secreted exoenzymes are less well understood, particularly for the sequenced strain PAO1. Therefore, we have systematically deleted the genes for the three known type III effector molecules (exoS, exoT, and exoY) in P. aeruginosa PAO1 and assayed the effect of the deletions, both singly and in combination, on cytotoxicity in vitro and in vivo. We found that the type III secretion system acts differently on different cell types, causing an exoST-dependent rounding of a lung epithelial-like cell line in contrast to causing an exoSTY-independent but translocase (popB)-dependent lysis of a macrophage cell line. We utilized an in vivo competitive infection model to test each of our mutants, examining replication in the lung and spread to secondary sites such as the blood and spleen. Type III mutants inoculated intranasally exhibited only a minor defect in replication and survival in the lung, but popB and exoSTY triple mutants were profoundly defective in their ability to spread systemically. Intravenous injection of the mutants indicated that the type III secretion machinery is required for survival in the blood. Furthermore, our findings suggest that the effector-independent popB-dependent cytotoxicity that we and others have observed in vitro in macrophage cell lines may not be of great importance in vivo.
机译：铜绿假单胞菌使用专用的III型分泌系统将毒素直接传递到宿主细胞的细胞质中。尽管在体外阐明III型分泌的毒素的功能方面已取得进展，但对于III型分泌的外切酶的体内功能仍知之甚少，尤其是对于测序菌株PAO1而言。因此，我们已系统删除了 P中三个已知的III型效应子分子（ exoS ， exoT 和 exoY ）的基因。 。铜绿假单胞菌PAO1，并单独或联合测定了这些缺失对体外和体内细胞毒性的影响。我们发现，III型分泌系统在不同细胞类型上的作用不同，与引起 exoSTY -的肺上皮样细胞系形成 exoST 依赖型舍入有关巨噬细胞系但不依赖转位酶（ popB ）的裂解。我们利用体内竞争性感染模型来测试我们的每个突变体，检查在肺中的复制并扩散到次要部位，例如血液和脾脏。经鼻内接种的III型突变体在肺中的复制和存活中仅表现出较小的缺陷，但是 popB 和 exoSTY 三重突变体在系统传播方面却存在严重缺陷。静脉内注射突变体表明III型分泌机制是血液中生存所必需的。此外，我们的发现提示我们和其他人在巨噬细胞系中体外观察到的与效应子无关的 popB 依赖性细胞毒性在体内可能并不重要。

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