In vitro model to estimate Edwardsiella tarda-macrophage interactions using RAW264.7 cells
In vitro model to estimate Edwardsiella tarda-macrophage interactions using RAW264.7 cells
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使用 RAW264.7 细胞估计迟缓爱德华氏菌-巨噬细胞相互作用的体外模型
DOI：
10.1016/j.fsi.2016.11.027
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发表时间：
2017-01-01
影响因子：
4.7
通讯作者：
Bi, Keran
中科院分区：
文献类型：
--
作者：
Qin, Lei;Sun, Yuying;Bi, Keran
迟缓爱德华氏菌是一种重要的兼性细胞内致病菌，能在巨噬细胞内存活和复制。E.迟发性巨噬细胞相互作用在鱼类抗感染的防御机制中起着非常重要的作用。探讨了E.用于感染巨噬细胞并在巨噬细胞内持续存在迟发型超敏反应还没有得到很好表征。为了深入了解这一过程，使用RAW 264.7细胞来研究E.迟缓和巨噬细胞。利用体外模型，涉及RAW 264.7细胞，内化实验表明，MOI为10：1和100：1可以导致令人满意的感染率后，2 h的感染期。与在鱼类巨噬细胞中的表现一致，E. tarda可在RAW264.7细胞中存活、复制并诱导iNOS介导的NO产生。光镜和电镜证实了E. RAW264.7细胞中的tarda，显示一旦进入巨噬细胞，许多细菌可以在吞噬溶酶体内被破坏，并且那些成功破坏吞噬细胞防御的细菌能够在巨噬细胞中的空泡样隔室内广泛复制。此外，E.在RAW264.7细胞中以剂量和时间依赖性方式观察到迟发性诱导的凋亡，其特征在于增加的膜联蛋白V结合和caspase-3的活化。本文所述的结果表明，RAW264.7细胞可以模拟鱼巨噬细胞对E. tarda在许多方面表现出不同的功能，可作为研究E.迟缓和巨噬细胞。E.迟发性侵袭RAW264.7细胞模型的建立为进一步研究E.迟发性发病机制(C)2016爱思唯尔有限公司版权所有。
Edwardsiella tarda has been recognized as an important facultative intracellular pathogen of fish with capability of survival and replication within macrophages. E. tarda-macrophage interactions play a very important role in the defense mechanism of fish against infection. The mechanisms that E. tarda use to infect and persist inside macrophages are not well characterized. To gain insight concerning this process, RAW264.7 cells was used to investigate the interactions between E. tarda and macrophages. Using an in vitro model involving RAW264.7 cells, internalization assay demonstrated that MOIs of 10:1 and 100:1 could result in a satisfactory infection rate after a 2 h infection period. Consistent with the performance in fish macrophages, E. tarda could survive, replicate and induce iNOS-mediated NO production in RAW264.7 cells. Light and electron microscopy confirmed the internalization and replication of E. tarda in RAW264.7 cells, showing once inside macrophages, numberous bacteria may be destroyed within phagolysosomes and those that successfully subvert phagocyte defenses are capable of extensively replicating within the vacuolar-like compartment in macrophages. In addition, E. tarda-induced apoptosis was observed in RAW264.7 cells in a dose-and time-dependent manner, characterized by increased Annexin V binding and the activation of caspase-3. The results described here indicate that RAW264.7 cells could model the behavior of fish macrophages in response to E. tarda in many ways and may serve as a cell model for study on interactions between E. tarda and macrophages. The successful establishment of E. tarda-invaded RAW264.7 cells model may contribute to providing a basis for more detailed understanding of E. tarda pathogenesis. (C) 2016 Elsevier Ltd. All rights reserved.