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基于实验结果,利用免疫信息学方法预测 cagW 中的潜在免疫优势表位,以探讨针对幽门螺杆菌感染的保护作用。

Immunoinformatic prediction of potential immunodominant epitopes from cagW in order to investigate protection against Helicobacter pylori infection based on experimental consequences.

机构信息

Novin Genome (NG) Lab, Research and Development Center for Biotechnology, Shahrekord, Iran.

Young Researchers and Elite Club, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran.

出版信息

Funct Integr Genomics. 2023 Mar 29;23(2):107. doi: 10.1007/s10142-023-01031-1.

Abstract

Helicobacter pylori is a leading cause of stomach cancer and peptic ulcers. Thus, identifying epitopes in H. pylori antigens is important for disease etiology, immunological surveillance, enhancing early detection tests, and developing optimal epitope-based vaccines. We used immunoinformatic and computational methods to create a potential CagW epitope candidate for H. pylori protection. The cagW gene of H. pylori was amplified and cloned into pcDNA3.1 (+) for injection into the muscles of healthy BALB/c mice to assess the impact of the DNA vaccine on interleukin levels. The results will be compared to a control group of mice that received PBS or cagW-pcDNA3.1 (+) vaccinations. An analysis of CagW protein antigens revealed 8 CTL and 7 HTL epitopes linked with AYY and GPGPG, which were enhanced by adding B-defensins to the N-terminus. The vaccine's immunogenicity, allergenicity, and physiochemistry were validated, and its strong activation of TLRs (1, 2, 3, 4, and 10) suggests it is antigenic. An in-silico cloning and immune response model confirmed the vaccine's expression efficiency and predicted its impact on the immune system. An immunofluorescence experiment showed stable and bioactive cagW gene expression in HDF cells after cloning the whole genome into pcDNA3.1 (+). In vivo vaccination showed that pcDNA3.1 (+)-cagW-immunized mice had stronger immune responses and a longer plasmid DNA release window than control-plasmid-immunized mice. After that, bioinformatics methods predicted, developed, and validated the three-dimensional structure. Many online services docked it with Toll-like receptors. The vaccine was refined using allergenicity, antigenicity, solubility, physicochemical properties, and molecular docking scores. Virtual-reality immune system simulations showed an impressive reaction. Codon optimization and in-silico cloning produced E. coli-expressed vaccines. This study suggests a CagW epitopes-protected H. pylori infection. These studies show that the proposed immunization may elicit particular immune responses against H. pylori, but laboratory confirmation is needed to verify its safety and immunogenicity.

摘要

幽门螺杆菌是胃癌和消化性溃疡的主要病因。因此,鉴定幽门螺杆菌抗原中的表位对于疾病病因学、免疫监测、增强早期检测试验以及开发最佳的基于表位的疫苗都很重要。我们使用免疫信息学和计算方法为幽门螺杆菌的保护创建了一个潜在的 CagW 表位候选物。扩增了幽门螺杆菌的 cagW 基因并将其克隆到 pcDNA3.1(+)中,以便注射到健康的 BALB/c 小鼠的肌肉中,以评估 DNA 疫苗对白细胞介素水平的影响。结果将与接受 PBS 或 cagW-pcDNA3.1(+)疫苗接种的对照组小鼠进行比较。对 CagW 蛋白抗原的分析显示,有 8 个 CTL 和 7 个 HTL 表位与 AYY 和 GPGPG 相关,通过在 N 端添加 B 防御素来增强其与抗原的结合。该疫苗的免疫原性、过敏性和物理化学性质得到了验证,其强烈激活 TLRs(1、2、3、4 和 10)表明其具有抗原性。通过计算机模拟克隆和免疫反应模型,证实了疫苗的表达效率,并预测了其对免疫系统的影响。免疫荧光实验表明,在将整个基因组克隆到 pcDNA3.1(+)后,HDF 细胞中 cagW 基因的表达稳定且具有生物活性。体内疫苗接种表明,pcDNA3.1(+)-cagW 免疫的小鼠比对照质粒免疫的小鼠具有更强的免疫反应和更长的质粒 DNA 释放窗口。之后,生物信息学方法对其进行了三维结构的预测、开发和验证。许多在线服务将其与 Toll 样受体对接。使用过敏性、抗原性、可溶性、物理化学性质和分子对接评分对疫苗进行了优化。虚拟免疫系统模拟显示出了令人印象深刻的反应。密码子优化和计算机模拟克隆产生了大肠杆菌表达的疫苗。本研究提出了一种 CagW 表位保护的幽门螺杆菌感染。这些研究表明,所提出的免疫接种可能会引起针对幽门螺杆菌的特定免疫反应,但需要实验室确认以验证其安全性和免疫原性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3915/10049908/6787b06c11e2/10142_2023_1031_Fig1_HTML.jpg

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