摘要
Abstract
Helicobacter pylori infection is a major global cause of gastritis,peptic ulcers and gastric cancer,presenting a huge public health challenge that urgently requires the development of effective vaccines.This study utilized an immunoinformatics approach to construct a novel multi-epitope vaccine through systematic epitope screening of key H.pylori virulence factors(OipA,VacA,IceA1,and HpaA).Based on stringent cri-teria(including antigenicity score＞0.4,non-allergenicity,non-toxicity,and no homology with human pro-teins),five highly immunogenic cytotoxic T lymphocyte(CTL;Tc cell)epitopes were selected from 173 candi-dates,six helper T cell(Th cell)epitopes were identified from 191 candidates,and eight linear B-cell epitopes were determined from 865 candidates.The selected epitopes were integrated with the LT-Ⅱ B adjuvant via linkers,resulting in a multi-epitope vaccine comprising 421 amino acid residues(molecular weight:44.613 kDa,theoretical pI:8.17).The vaccine exhibited high antigenicity(score:1.096 4),stability and hydrophilicity.Structural analysis revealed a well-proportioned distribution of secondary structures in the vaccine:37.055％α-helices,7.363％β-turns,38.242％random coils,and 17.340％extended strands.Molecular docking con-firmed stable binding interactions between the vaccine and Toll-like receptors(TLR2,TLR4,TLR5,TLR9 and TLR10),suggesting its potential to effectively activate innate immunity.Immunosimulation analysis de-monstrated that this vaccine can elicit robust humoral and cellular immune responses,with theoretical popu-lation coverage of 76.18％.Furthermore,the codon-optimized vaccine sequence was successfully cloned into the pET-28a(+)vector,laying the foundation for subsequent experiments.In summary,the constructed multi-epitope vaccine exhibited both broad-spectrum protective potential and high safety.This study provides an innovative design strategy for H.pylori vaccine development,and also offers a reference model for vaccines against other pathogens.关键词
幽门螺杆菌/多表位疫苗/免疫信息学/抗生素耐药性Key words
Helicobacter pylori/multi-epitope vaccine/immunoinformatics/antibiotic resistance分类
生物工程
