Luis Fang, Dalgys Martínez, Catherine Meza-Torres, Ana Moreno-Woo, Nicole Pereira-Sanandrés, Alex Domínguez Vargas, Gloria Garavito, Eduardo Egea
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引用次数: 0
Abstract
Objective: This study aimed to identify by in silico methods tropomyosin consensus B and T epitopes of shrimp species, house dust mites, insects, and nematodes associated with allergic diseases in tropical countries.
Methods: In silico analysis included tropomyosin from mites (Der p 10, Der f 10, Blo t 10), insects (Aed a 10, Per a 7, Bla g 7), shrimp (Lit v 1, Pen m 1, Pen a 1), and nematode (Asc l 3) all sequences were taken from the UniProt database. Linear IgE epitopes were predicted with AlgPred 2.0 and validated with BepiPred 3.0. MHC-II binding T cell epitopes were predicted using the IEDB server, which implements nine predictive methods (consensus method, combinatorial library, NN-align-2.3, NN- align-2.2, SMM-align, Sturniolo, NetMHCIIpan 3.1, and NetMHCIIpan 3.2) these predictions focused on 10 HLA-DR and 2 HLA-DQ alleles associated with allergic diseases. Subsequently, consensus B and T epitopes present in all species were identified.
Results: We identified 12 sequences that behaved as IgE-epitopes and B-cell epitopes, three of them: 160RKYDEVARKLAMVEA174, 192ELEEELRVVGNNLKSLEVSEEKAN215, 251KEVDRLEDELV261 were consensus in all species. Eleven peptides (T-epitopes) showed strong binding (percentile rank ≤ 2.0) to HLA-DRB1*0301, *0402, *0411, *0701, *1101, *1401, HLA-DQA1*03:01/DQB1*03:02, and HLA- DQA1*05:01/DQB1*02:01. Only two T-epitopes were consensus in all species: 167RKLAMVEADLERAEERAEt GEsKIVELEEELRV199, and 218EEeY KQQIKT LTaKLKEAEARAEFAERSV246. Subsequently, we identified 2 B and T epitope sequences and reached a consensus between species 167RKLAMVEA174 and 192ELEEELRV199.
Conclusions: These data describe three sequences that may explain the IgE cross-reactivity between the analyzed species. In addition, the consensus B and T epitopes can be used for further in vitro investigations and may help to design multiple-epitope protein-based immunotherapy for tropomyosin-related allergic diseases.
目的:本研究旨在通过硅学方法确定与热带国家过敏性疾病相关的虾类、屋尘螨、昆虫和线虫的肌球蛋白共识 B 和 T 表位:本研究旨在通过硅学方法确定与热带国家过敏性疾病相关的虾类、屋尘螨、昆虫和线虫的肌球蛋白共识 B 和 T 表位:硅学分析包括来自螨虫(Der p 10、Der f 10、Blo t 10)、昆虫(Aed a 10、Per a 7、Bla g 7)、虾(Lit v 1、Pen m 1、Pen a 1)和线虫(Asc l 3)的肌球蛋白,所有序列均来自 UniProt 数据库。线性 IgE 表位用 AlgPred 2.0 预测,并用 BepiPred 3.0 验证。MHC-II结合T细胞表位是用IEDB服务器预测的,该服务器实现了九种预测方法(共识法、组合库、NN-align-2.3、NN- align-2.2、SMM-align、Sturniolo、NetMHCIIpan 3.1和NetMHCIIpan 3.2),这些预测主要针对与过敏性疾病相关的10个HLA-DR和2个HLA-DQ等位基因。随后,确定了存在于所有物种中的共识 B 和 T 表位:结果:我们确定了 12 个可作为 IgE 表位和 B 细胞表位的序列,其中三个序列是160RKYDEVARKLAMVEA174, 192ELEEELRVVGNNLKSLEVSEEKAN215, 251KEVDRLEDELV261 是所有物种的共识。有 11 条肽(T 表位)与 HLA-DRB1*0301、*0402、*0411、*0701、*1101、*1401、HLA-DQA1*03:01/DQB1*03:02 和 HLA- DQA1*05:01/DQB1*02:01 有强结合(百分位数≤ 2.0)。在所有物种中,只有两个 T 表位是一致的:167RKLAMVEADLERAEERAEt GEsKIVELEEELRV199 和 218EEeY KQQIKT LTaKLKEAEARAEFAERSV246。随后,我们确定了 2 个 B 和 T 表位序列,并在物种 167RKLAMVEA174 和 192ELEEELRV199 之间达成了共识:这些数据描述了三个序列,它们可能解释了所分析物种之间的 IgE 交叉反应。此外,共识的 B 和 T 表位可用于进一步的体外研究,并有助于设计基于多表位蛋白的免疫疗法来治疗肌球蛋白相关的过敏性疾病。