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兔宿主对热灭活土拉弗朗西斯菌活疫苗株抗体反应的免疫蛋白质组学分析

Immunoproteomic Analysis of Antibody Response of Rabbit Host Against Heat-Killed Francisella tularensis Live Vaccine Strain.

作者信息

Gaur Ritu, Alam Syed Imteyaz, Kamboj Dev Vrat

机构信息

Biotechnology Division, Defence Research and Development Establishment, Gwalior, 474002, India.

出版信息

Curr Microbiol. 2017 Apr;74(4):499-507. doi: 10.1007/s00284-017-1217-y. Epub 2017 Feb 23.

Abstract

Francisella tularensis, the causative agent of tularemia, has attained the status of one of the high priority agents that could be used in the act of bioterrorism. Currently, there is no licensed vaccine for this highly infectious intracellular pathogen. Being a listed 'Category A' agent of the U.S. Center for Disease Control and Prevention (CDC), vaccines and therapeutics are immediately required against this pathogen. In this study, an immunoproteomic approach based on the techniques of 2-dimensional gel electrophoresis (2DE) and immunoblotting combined with mass spectrometry (MS) was used for elucidation of immunogenic components and putative vaccine candidates. Whole-cell soluble protein extract of F. tularensis LVS (Ft LVS) was separated by 2DE, and immunoblots were developed with sera raised in rabbit after immunization with heat-killed Ft LVS. A total of 28 immunoreactive proteins were identified by tandem mass spectrometry. Rabbit immunoproteome of F. tularensis was compared with those previously reported using sera from human patients and in murine model. Out of 28 immunoreactive proteins identified in this study, 12 and 17 overlapping proteins were recognized by human and murine sera, respectively. Nine proteins were found immunogenic in all the three hosts, while eight new immunogenic proteins were found in this study. Identified immunoreactive proteins may find application in design and development of protein subunit vaccine for tularemia.

摘要

土拉弗朗西斯菌是兔热病的病原体,已成为可用于生物恐怖主义行为的高优先级病原体之一。目前,针对这种具有高度传染性的细胞内病原体尚无获批的疫苗。作为美国疾病控制与预防中心(CDC)列出的“A类”病原体,迫切需要针对该病原体的疫苗和治疗方法。在本研究中,采用了基于二维凝胶电泳(2DE)和免疫印迹技术并结合质谱(MS)的免疫蛋白质组学方法来阐明免疫原性成分和推定的疫苗候选物。用二维凝胶电泳分离土拉弗朗西斯菌LVS(Ft LVS)的全细胞可溶性蛋白提取物,并用热灭活的Ft LVS免疫兔子后产生的血清进行免疫印迹分析。通过串联质谱法共鉴定出28种免疫反应性蛋白。将土拉弗朗西斯菌的兔免疫蛋白质组与先前使用人类患者血清和小鼠模型报道的免疫蛋白质组进行比较。在本研究中鉴定出的28种免疫反应性蛋白中,分别有12种和17种重叠蛋白被人类和小鼠血清识别。发现有9种蛋白在所有三种宿主中均具有免疫原性,而在本研究中发现了8种新的免疫原性蛋白。鉴定出的免疫反应性蛋白可能在兔热病蛋白质亚单位疫苗的设计和开发中得到应用。

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