通过蛋白质组学和肽组学鉴定布鲁氏菌潜在的抗原肽。
Identification of potential antigenic peptides of Brucella through proteome and peptidome.
机构信息
Department of Clinical Laboratory, The Third Medical Centre of Chinese PLA General Hospital, The Training Site for Postgraduate of Jinzhou Medical University, Beijing, China.
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology, Beijing, China.
出版信息
Vet Med Sci. 2023 Jan;9(1):523-534. doi: 10.1002/vms3.1048. Epub 2022 Dec 30.
BACKGROUND
Brucellosis, caused by Brucella spp., is a major zoonotic public health threat. Although several Brucella vaccines have been demonstrated for use in animals, Brucella spp. can cause human infection and to date, there are no human-use vaccines licensed by any agency. Recently, methods in vaccine informatics have made major breakthroughs in peptide-based epitopes, opening up a new avenue of vaccine development.
OBJECTIVES
The purpose of this article was to identify potential antigenic peptides in Brucella by proteome and peptidome analyses.
METHODS
Mouse infection models were first established by injection with Brucella and spleen protein profiles were then analysed. Subsequently, the major histocompatibility complex class I or II (major histocompatibility complex [MHC]-I/II)-binding peptides in blood samples were collected by immunoprecipitation and peptides derived from Brucella proteins were identified through liquid chromatography-mass spectrometry (LC-MS/MS). These peptides were then evaluated in a variety of ways, such as in terms of conservation in Brucella and synchronicity in predicted peptides (similarity and coverage), which allowed us to more effectively measure their antigenic potential.
RESULTS
The expression of the inflammatory cytokines IL1B and IFN-γ was significantly altered in the spleen of infected mice and some Brucella proteins, such as Muri, AcpP and GroES, were also detected. Meanwhile, in blood, 35 peptides were identified and most showed high conservation, highlighting their potential as antigen epitopes for vaccine development. In particular, we identified four proteins containing both MHC-I- and MHC-II-binding peptides including AtpA, AtpD, DnaK and BAbS19_II02030. They were also compared with the predicted peptides to estimate their reliability.
CONCLUSIONS
The peptides we screened could bind to MHC molecules. After being stimulated with antigen T epitopes, Memory T cells can stimulate T cell activation and promote immune responses. Our results indicated that the peptides we identified may be good candidate targets for the design of subunit vaccines and these results pave the way for the study of safer vaccines against Brucella.
背景
布鲁氏菌病是由布鲁氏菌属引起的一种主要的人畜共患公共卫生威胁。虽然已经有几种布鲁氏菌疫苗被证明可用于动物,但布鲁氏菌属仍可引起人类感染,迄今为止,还没有任何机构批准用于人类的疫苗。最近,疫苗信息学方法在肽基表位方面取得了重大突破,为疫苗开发开辟了新途径。
目的
本文旨在通过蛋白质组和肽组分析鉴定布鲁氏菌中的潜在抗原肽。
方法
首先通过注射布鲁氏菌建立小鼠感染模型,然后分析脾脏蛋白谱。随后,通过免疫沉淀从血液样本中收集主要组织相容性复合体 I 类或 II 类(主要组织相容性复合体 [MHC]-I/II)结合肽,并通过液相色谱-质谱联用(LC-MS/MS)鉴定来自布鲁氏菌蛋白的肽。然后通过多种方式评估这些肽,例如在布鲁氏菌中的保守性和预测肽的同步性(相似性和覆盖率),这使我们能够更有效地衡量它们的抗原潜力。
结果
感染小鼠脾脏中炎症细胞因子 IL1B 和 IFN-γ 的表达明显改变,同时也检测到一些布鲁氏菌蛋白,如 Muri、AcpP 和 GroES。此外,在血液中鉴定出 35 个肽,大多数具有高度保守性,这突出了它们作为疫苗开发抗原表位的潜力。特别是,我们鉴定了四个含有 MHC-I 和 MHC-II 结合肽的蛋白质,包括 AtpA、AtpD、DnaK 和 BAbS19_II02030。我们还将它们与预测的肽进行比较,以估计它们的可靠性。
结论
我们筛选的肽可以与 MHC 分子结合。经过抗原 T 表位刺激后,记忆 T 细胞可以刺激 T 细胞活化并促进免疫反应。我们的研究结果表明,我们鉴定的肽可能是设计亚单位疫苗的良好候选靶标,这些结果为研究更安全的布鲁氏菌疫苗铺平了道路。