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噬菌体在肠出血性大肠杆菌感染中的作用:对预防和治疗方法设计的新启示

Role of bacteriophages in STEC infections: new implications for the design of prophylactic and treatment approaches.

作者信息

Amorim Jaime H, Del Cogliano Manuel E, Fernandez-Brando Romina J, Bilen Marcos F, Jesus Monica R, Luiz Wilson B, Palermo Marina S, Ferreira Rita C C, Servat Esteban G, Ghiringhelli Pablo D, Ferreira Luis C S, Bentancor Leticia V

机构信息

Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, 05508-060, Brazil.

Laboratorio de Ingeniería Genética y Biología Celular y Molecular, Universidad Nacional de Quilmes, Buenos Aires, 1876, Argentina.

出版信息

F1000Res. 2014 Mar 18;3:74. doi: 10.12688/f1000research.3718.2. eCollection 2014.

Abstract

Shiga toxin (Stx) is considered the main virulence factor in Shiga toxin-producing Escherichia coli (STEC) infections. Previously we reported the expression of biologically active Stx by eukaryotic cells in vitro and in vivo following transfection with plasmids encoding Stx under control of the native bacterial promoter (1,2). Since stx genes are present in the genome of lysogenic bacteriophages, here we evaluated the relevance of bacteriophages during STEC infection. We used the non-pathogenic E. coli C600 strain carrying a lysogenic 933W mutant bacteriophage in which the stx operon was replaced by a gene encoding the green fluorescent protein (GFP). Tracking GFP expression using an In Vivo Imaging System (IVIS), we detected fluorescence in liver, kidney, and intestine of mice infected with the recombinant E. coli strain after treatment with ciprofloxacin, which induces the lytic replication and release of bacteriophages. In addition, we showed that chitosan, a linear polysaccharide composed of d-glucosamine residues and with a number of commercial and biomedical uses, had strong anti-bacteriophage effects, as demonstrated at in vitro and in vivo conditions. These findings bring promising perspectives for the prevention and treatment of haemolytic uremic syndrome (HUS) cases.

摘要

志贺毒素(Stx)被认为是产志贺毒素大肠杆菌(STEC)感染中的主要毒力因子。此前我们报道,在用天然细菌启动子控制下编码Stx的质粒转染后,真核细胞在体外和体内均可表达具有生物活性的Stx(1,2)。由于stx基因存在于溶原性噬菌体的基因组中,因此我们在此评估了噬菌体在STEC感染过程中的相关性。我们使用了携带溶原性933W突变噬菌体的非致病性大肠杆菌C600菌株,其中stx操纵子被编码绿色荧光蛋白(GFP)的基因所取代。使用体内成像系统(IVIS)追踪GFP表达,在用环丙沙星处理后,我们在感染重组大肠杆菌菌株的小鼠的肝脏、肾脏和肠道中检测到荧光,环丙沙星可诱导噬菌体的裂解复制和释放。此外,我们还表明,壳聚糖是一种由d-葡萄糖胺残基组成的线性多糖,具有多种商业和生物医学用途,在体外和体内条件下均显示出强大的抗噬菌体作用。这些发现为溶血尿毒综合征(HUS)病例的预防和治疗带来了有希望的前景。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7b2f/4288508/f4d5fb2c17af/f1000research-3-5299-g0000.jpg

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