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免疫信息学方法鉴定的用于控制鲍曼不动杆菌的新型外膜抗原多表位 DNA 疫苗候选物。

Putative novel outer membrane antigens multi-epitope DNA vaccine candidates identified by Immunoinformatic approaches to control Acinetobacter baumannii.

机构信息

Department of Biology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran.

Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran.

出版信息

BMC Immunol. 2023 Nov 18;24(1):46. doi: 10.1186/s12865-023-00585-w.

Abstract

Multi-epitope polypeptide vaccines, a fusion protein, often have a string-of-beads system composed of various specific peptide epitopes, potential adjuvants, and linkers. When choosing the sequence of various segments and linkers, many alternatives are available. These variables can influence the vaccine's effectiveness through their effects on physicochemical properties and polypeptide tertiary structure.The most conserved antigens were discovered using BLASTn. To forecast the proteins' subcellular distribution, PSORTb 3.0.2 was used. Vaxign was used for the preliminary screening and antigenicity assessment. Protein solubility was also predicted using the ccSOL omics. Using PRED-TMBB, it was anticipated that the protein would localize across membranes. The IEDB and BepiPred-2.0 databases were used to predict the immunogenicity of B cell epitopes. A multi-epitope construct was developed and analyzed to evaluate. Twenty epitopes from A. baumannii's outer membrane protein (omp) were included in the vaccination. TLR4 agonist explosibility was investigated. The physicochemical characteristics, secondary and tertiary structures, and B-cell epitopes of vaccine constructs were assessed. Additionally, docking and MD experiments were used to examine the relationship between TLR4 and its agonist.Thirteen antigens were discovered, and eight of the 13 chosen proteins were predicted to be surface proteins. The 34 kDa outer membrane protein, Omp38, Omp W, CarO, putative porin, OmpA, were chosen as having the right antigenicity (≥0.5). FhuE and CdiA were eliminated from further study because of their low antigenicity. The vaccine design was developed by combining the most effective 10 B-cell and 10 MHC-I/MHCII combined coverage epitopes. The molecular formula of the vaccine was determined to be C1718H2615N507O630S17. The vaccine form has a molecular weight of 40,996.70 Da and 47 negatively charged residues (Asp + Glu), whereas 28 positively charged residues (Arg + Lys). The estimated half-life was 7.2 hours (mammalian reticulocytes, in vitro), > 20 hours (yeast, in vivo) and > 10 hours (Escherichia coli, in vivo) for the vaccine. The multi-epitope vaccine insertion is carried via the expression vector pcDNA3.1 (+).The multi-epitope vaccine may stimulate humoral and cellular immune responses, according to our findings, and it may be a candidate for an A. baumannii vaccine.

摘要

多表位多肽疫苗,一种融合蛋白,通常具有由各种特定肽表位、潜在佐剂和连接子组成的串珠系统。在选择各种片段和连接子的序列时,有许多可供选择的方案。这些变量可以通过影响理化性质和多肽三级结构来影响疫苗的有效性。使用 BLASTn 发现最保守的抗原。为了预测蛋白质的亚细胞分布,使用了 PSORTb 3.0.2。Vaxign 用于初步筛选和抗原性评估。使用 ccSOL omics 预测蛋白质的溶解度。使用 PRED-TMBB 预测蛋白质跨膜定位。使用 IEDB 和 BepiPred-2.0 数据库预测 B 细胞表位的免疫原性。开发并分析了一种多表位构建体进行评估。包含了来自鲍曼不动杆菌外膜蛋白(omp)的 20 个表位。研究了 TLR4 激动剂的爆炸性。评估了疫苗构建体的理化特性、二级和三级结构以及 B 细胞表位。此外,还进行了对接和 MD 实验以研究 TLR4 与其激动剂之间的关系。发现了 13 种抗原,其中 13 种选定的蛋白质中有 8 种被预测为表面蛋白。34 kDa 外膜蛋白 Omp38、OmpW、CarO、假定孔蛋白 OmpA 被选为具有适当抗原性(≥0.5)。由于抗原性低,FhuE 和 CdiA 被排除在进一步研究之外。通过结合最有效的 10 个 B 细胞和 10 个 MHC-I/MHCII 结合覆盖表位来设计疫苗。确定疫苗的分子式为 C1718H2615N507O630S17。疫苗形式的分子量为 40996.70 Da,带 47 个负电荷残基(Asp+Glu),带 28 个正电荷残基(Arg+Lys)。疫苗在哺乳动物网织红细胞中的半衰期估计为 7.2 小时(体外),在酵母中的半衰期估计为 >20 小时(体内),在大肠杆菌中的半衰期估计为 >10 小时(体内)。多表位疫苗通过表达载体 pcDNA3.1(+)进行插入。根据我们的研究结果,多表位疫苗可能会刺激体液和细胞免疫反应,它可能是鲍曼不动杆菌疫苗的候选者。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6692/10657578/2dc7a8986ba0/12865_2023_585_Fig1_HTML.jpg

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