Construction of a Multi-epitope Vaccine Against Salmonella Pullorum by Immunoinformatics Approach
[Objective]This study was aimed to design multi-epitope vaccine against IpaJ protein of Salmonella Pullorum and provide a new vaccine for purifying chicken dysentery.[Method]In this study,IEDB was used to predict the major histocompatibility complex class Ⅰ(MHC Ⅰ)molecular binding epitopes of Salmonella Pullorum IpaJ protein in T lymphocytes.NetMHCIIpan 4.0 Server was used to predict T lymphocytes MHC Ⅱ molecular binding epitopes.B lymphocyte epitope was predicted by IEDB.After the antigenicity of the selected epitopes was evaluated by VaxiJen v 2.0,the qualified epitopes were concatenated into multi-epitope vaccine by flexible linker.Antigenicity,physicochemical properties,N-glycosylation sites,secondary structure and tertiary structure of the constructed multi-epitope vaccine were predicted.Molecular docking was used to evaluate the binding ability of the multi-epitope vaccine to the immune receptor.Immune simulation was used to evaluate the immune effect of the multi-epitope vaccine.Finally,the codon was optimized for cloning expression.[Result]After screening,the constructed multi-epitope vaccine MEV-IpaJ contained 4 MHC Ⅰ,4 MHC Ⅱ and 8 B lymphocyte epitope dominance.The relative molecular weight of the multi-epitope vaccine MEV-IpaJ was 28.18 ku,which was a stable hydrophilic protein and had good antigenicity.There were four potential N-glycoylation sites,including a-helix,extended strand,beta turn and random coil accounted for 20.38%,19.23%,8.08%and 52.31%,respectively.Ramachandran mapping of tertiary structure showed that the dominant region contained 89.9%of the residual base,and the residual base of the dominant region increased to 94.0%after refinement.The mapping of tertiary structure prominent epitope also proved that the multi-epitope vaccine had good immunogenicity,and molecular docking showed that the MEV-IpaJ could dock with Toll-like receptor 2(TLR2)and TLR4 protein molecules.The results of immune simulation showed that the MEV-IpaJ had a good immune response and could improve the expression of some cytokines.Codon optimization ensured the efficient and stable expression of the MEV-IpaJ in E.coli K12 expression system.[Conclusion]The constructed multi-epitope vaccine MEV-IpaJ could be effectively expressed and might induce strong T cell and B cell immune responses.This study provided a new method for the design of multi-epitope vaccine of Salmonella Pullorum,and provided theoretical basis and data support for the research and development of multi-epitope vaccine of Salmonella Pullorum.