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首页> 外文期刊>Parasitology >Functional evaluation of gene silencing on macrophages derived from U937 cells using interference RNA (shRNA) in a model of macrophages infected with Leishmania (Viannia) braziliensis.
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Functional evaluation of gene silencing on macrophages derived from U937 cells using interference RNA (shRNA) in a model of macrophages infected with Leishmania (Viannia) braziliensis.

机译：使用干扰RNA（ShRNA）在巨噬细胞模型中衍生自U937细胞巨噬细胞基因沉默的功能评估嗜血糖（Viannia）Braziliensis。

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摘要

Leishmaniasis development is multifactorial; nonetheless, the establishment of the infection, which occurs by the survival and replication of the parasite inside its main host cell, the macrophage, is mandatory. Thus, the importance of studying the molecular mechanisms involved in the Leishmania-macrophage interaction is highlighted. The aim of this study was to characterize a cellular model of macrophages derived from U937 cells that would allow for the identification of infection phenotypes induced by genetic silencing with interference RNA in the context of macrophages infected with Leishmania (Viannia) braziliensis. The model was standardized by silencing an exogenous gene (gfp), an endogenous gene (lmna) and a differentially expressed gene between infected and non-infected macrophages (gro-β). The silencing process was successful for the three genes studied, obtaining reductions of 88·9% in the GFP levels, 87·5% in LMNA levels and 74·4% for Gro-β with respect to the corresponding control cell lines. The cell model revealed changes in the infection phenotype of the macrophages in terms of number of amastigotes per infected macrophage, number of amastigotes per sampled macrophage and percentage of infected macrophages as a result of gene silencing. Thus, this cell model constitutes a research platform for the study of parasite-host interactions and for the identification of potentially therapeutic targets.
机译：利什曼病的发展是多因素;尽管如此，感染的建立，通过寄生虫在其主要宿主细胞内的存活率和复制发生，巨噬细胞是强制性的。因此，突出了研究嗜血巨噬细胞相互作用的分子机制的重要性。本研究的目的是表征来自U937细胞的巨噬细胞的细胞模型，该细胞模型将允许鉴定通过在感染利什曼（viannnia）巴西的巨噬细胞的巨噬细胞的遗传沉默遗传沉默诱导的感染表型。通过沉默外源基因（GFP），内源基因（LMNA）和感染的未感染和未感染的巨噬细胞（GRO-β）之间的差异表达基因来标准化该模型。沉默方法对于所研究的三个基因成功，在GFP水平中获得88·9％的减少，LMNA水平87·5％，对于相应的对照细胞系，Gro-β的74·4％。细胞模型在每次受感染的巨噬细胞的Amastigotes的数量方面揭示了巨噬细胞的感染表型的变化，由于基因沉默，每次取样巨噬细胞的巨噬细胞数和感染巨噬细胞的百分比。因此，该细胞模型构成了用于研究寄生虫 - 宿主相互作用的研究平台，并用于鉴定潜在的治疗目标。

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