基于减法蛋白组学和反向疫苗学方法设计禽致病性大肠杆菌O1和O78血清型多表位疫苗

doi:10.16431/j.cnki.1671-7236.2024.12.029

Abstract

Abstract: 【Objective】 This study was aimed to design a multi-epitope vaccine (MEV) against avian pathogenic Escherichia coli (APEC) O1 and O78 serotypes,laying a foundation for the development of new vaccines for APEC.【Method】 This study combined subtraction proteomics and reverse vaccinology.Redundant and non-similar proteins in the protein sequences of APEC O1 and O78 were removed using CD-HIT and BLASTP tools.Similar proteins in APEC O1 and O78 were removed by BLASTP and compared with reference proteomes of chicken,retaining non-homologous proteins.Essential toxigenic proteins were screened using DEG,VFDB,etc.databases.Candidate proteins were selected using PSORTb and VaxiJen v 2.0.T-cell major histocompatibility complex (MHC) class Ⅰ and Ⅱ molecule-binding epitopes were predicted using NetCTL 1.2 and NetMHCⅡpan 4.0,and B-cell epitopes were predicted using IEDB.Multi-epitope vaccine were evaluated for antigenicity by VaxiJen v 2.0,and the qualified epitopes were linked together by flexible linkers to form a multi-epitope vaccine.The antigenicity,physical and chemical properties,N-glycosylation sites,secondary structure and tertiary structure of the constructed multi-epitope vaccine were predicted.The binding ability of the multi-epitope vaccine to immune receptors was evaluated by molecular docking,and the immune effect was evaluated by immune simulation.Finally,the codons were optimized for cloning and expression.【Result】 After screening,12 MHC Ⅰ,12 MHC Ⅱ and 12 B lymphocyte epitope-dominant epitopes were selected to construct the multi-epitope vaccine MEV-O1O78.The molecular mass of the multi-epitope vaccine MEV-O1O78 was 69.81 ku,a stable hydrophilic protein with good antigenicity,containing 7 potential N-glycosylation sites.In the secondary structure,alpha-helix,extended chain and random coil accounted for 7.93%,10.81% and 81.27%,respectively.The Ramachandran plot of the tertiary structure showed that the epitope-rich region contained 95.6% of the residues.The immune simulation results showed that multi-epitope MEV-O1O78 could induce good humoral immunity and enhance the expression of some cytokines.The codon optimization ensured that the designed multi-epitope MEV-O1O78 was efficiently and stably expressed in the Escherichia coli K12 expression system.【Conclusion】 In this study,the APEC O1 and O78 multi-epitope vaccine MEV-O1O78 containing 36 dominant epitopes was successfully designed,which provided theoretical basis and data support for the development of multi-epitope vaccine for avian pathogenic colibacillosis.

Key words: avian pathogenic Escherichia coli (APEC); serotype; multi-epitope vaccine

CLC Number:

S852.4

CHEN Hong, WU Shuang, CHE Yegui, QIU Shulei, ZHOU Zixiang, WANG Yi, WANG Yongjuan, YUAN Cheng. Design of Multi-epitope Vaccines for Avian Pathogenic Escherichia coli O1 and O78 Serotypes Based on Subtractive Proteomics and Reverse Vaccinology[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(12): 5425-5438.

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Design of Multi-epitope Vaccines for Avian Pathogenic Escherichia coli O1 and O78 Serotypes Based on Subtractive Proteomics and Reverse Vaccinology

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