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结合蛋白质组学和生物信息学探索新的囊虫抗原作为抗细粒棘球蚴感染的疫苗候选物。

Combining proteomics and bioinformatics to explore novel tegumental antigens as vaccine candidates against Echinococcus granulosus infection.

机构信息

Área Inmunología, DEPBIO/IQB, Facultad de Química/Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay.

Unidad de Bioquímica y Proteómica Analíticas, Institut Pasteur de Montevideo and IIBCE, Montevideo, Uruguay.

出版信息

J Cell Biochem. 2019 Sep;120(9):15320-15336. doi: 10.1002/jcb.28799. Epub 2019 Apr 30.

Abstract

Echinococcus granulosus is the parasite responsible for cystic echinococcosis (CE), an important worldwide-distributed zoonosis. New effective vaccines against CE could potentially have great economic and health benefits. Here, we describe an innovative vaccine design scheme starting from an antigenic fraction enriched in tegumental antigens from the protoscolex stage (termed PSEx) already known to induce protection against CE. We first used mass spectrometry to characterize the protein composition of PSEx followed by Gene Ontology analysis to study the potential Biological Processes, Molecular Functions, and Cellular Localizations of the identified proteins. Following, antigenicity predictions and determination of conservancy degree against other organisms were determined. Thus, nine novel proteins were identified as potential vaccine candidates. Furthermore, linear B cell epitopes free of posttranslational modifications were predicted in the whole PSEx proteome through colocalization of in silico predicted epitopes within peptide fragments identified by matrix-assisted laser desorption/ionization-TOF/TOF. Resulting peptides were termed "clean linear B cell epitopes," and through BLASTp scanning against all nonhelminth proteins, those with 100% identity against any other protein were discarded. Then, the secondary structure was predicted for peptides and their corresponding proteins. Peptides with highly similar secondary structure respect to their parental protein were selected, and those potentially toxic and/or allergenic were discarded. Finally, the selected clean linear B cell epitopes were mapped within their corresponding 3D-modeled protein to analyze their possible antibody accessibilities, resulting in 14 putative peptide vaccine candidates. We propose nine novel proteins and 14 peptides to be further tested as vaccine candidates against CE.

摘要

细粒棘球蚴是引起包虫病(CE)的寄生虫,这是一种在世界范围内广泛分布的重要动物源性传染病。针对 CE 的新型有效疫苗可能具有巨大的经济和健康效益。在这里,我们描述了一种从已被证明可诱导对 CE 产生保护作用的原头蚴阶段富含表皮抗原的抗原性片段(称为 PSEx)开始的创新疫苗设计方案。我们首先使用质谱法对 PSEx 的蛋白质组成进行了表征,然后使用基因本体论分析研究了鉴定蛋白的潜在生物学过程、分子功能和细胞定位。接下来,预测了抗原性并确定了对其他生物体的保守程度。因此,鉴定了 9 种新的蛋白质作为潜在的疫苗候选物。此外,通过在通过基质辅助激光解吸/电离-TOF/TOF 鉴定的肽片段内的计算预测的表位与体内预测的表位的共定位,在整个 PSEx 蛋白质组中预测了无翻译后修饰的线性 B 细胞表位。所得肽被称为“清洁线性 B 细胞表位”,并通过 BLASTp 扫描针对所有非寄生虫蛋白,排除与任何其他蛋白质具有 100%同一性的那些。然后,对肽及其相应的蛋白质进行二级结构预测。选择与亲本蛋白具有高度相似二级结构的肽,并排除那些潜在有毒和/或致敏的肽。最后,将选定的清洁线性 B 细胞表位映射到其相应的 3D 建模蛋白中,以分析它们可能的抗体可及性,从而得到 14 个潜在的肽疫苗候选物。我们提出了 9 种新蛋白和 14 个肽作为针对 CE 的疫苗候选物进行进一步测试。

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