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从水中分离T5:对其致病潜力的表征,包括细胞外囊泡的产生

Isolation of T5 from Water: Characterization of Its Pathogenic Potential, Including the Production of Extracellular Vesicles.

作者信息

Retana Moreira Lissette, Vargas Ramírez Daniel, Linares Fátima, Prescilla Ledezma Alexa, Vaglio Garro Annette, Osuna Antonio, Lorenzo Morales Jacob, Abrahams Sandí Elizabeth

机构信息

Departamento de Parasitología, Universidad de Costa Rica, San Pedro, Montes de Oca 2060, Costa Rica.

Centro de Investigación en Enfermedades Tropicales (CIET), Universidad de Costa Rica, San Pedro, Montes de Oca 2060, Costa Rica.

出版信息

Pathogens. 2020 Feb 21;9(2):144. doi: 10.3390/pathogens9020144.

Abstract

is a genus of free-living amoebae widely distributed in nature, associated with the development of encephalitis and keratitis. Despite the fact that it is common to find genotype T5 in environmental samples, only a few cases have been associated with clinical cases in humans. The wide distribution of , the characteristic of being amphizoic and the severity of the disease motivate researchers to focus on the isolation of these organisms, but also in demonstrating direct and indirect factors that could indicate a possible pathogenic potential. Here, we performed the characterization of the pathogenic potential of an T5 isolate collected from a water source in a hospital. Osmo- and thermotolerance, the secretion of proteases and the effect of trophozoites over cell monolayers were analyzed by different methodologies. Additionally, we confirm the secretion of extracellular vesicles (EVs) of this isolate incubated at two different temperatures, and the presence of serine and cysteine proteases in these vesicles. Finally, using atomic force microscopy, we determined some nanomechanical properties of the secreted vesicles and found a higher value of adhesion in the EVs obtained at 37 °C, which could have implications in the parasite´s survival and damaging potential in two different biological environments.

摘要

是一种自由生活的变形虫属,广泛分布于自然界,与脑炎和角膜炎的发生有关。尽管在环境样本中常见基因型T5,但仅有少数病例与人类临床病例相关。其广泛分布、兼性特征以及疾病的严重性促使研究人员不仅关注这些生物体的分离,还关注证明可能表明潜在致病潜力的直接和间接因素。在此,我们对从医院水源采集的一株T5分离株的致病潜力进行了表征。通过不同方法分析了渗透压和耐热性、蛋白酶分泌以及滋养体对细胞单层的影响。此外,我们证实了该分离株在两种不同温度下孵育时细胞外囊泡(EVs)的分泌,以及这些囊泡中丝氨酸和半胱氨酸蛋白酶的存在。最后,使用原子力显微镜,我们测定了分泌囊泡的一些纳米力学性质,发现在37℃获得的EVs中粘附力更高,这可能对寄生虫在两种不同生物环境中的存活和破坏潜力有影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6615/7168589/58544dbc058e/pathogens-09-00144-g001.jpg

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