Development of a multi-epitope peptide vaccine inducing robust T cell responses against brucellosis using immunoinformatics based approaches

Infect Genet Evol. 2017 Jul:51:227-234. doi: 10.1016/j.meegid.2017.04.009. Epub 2017 Apr 11.

Abstract

Current investigations have demonstrated that a multi-epitope peptide vaccine targeting multiple antigens could be considered as an ideal approach for prevention and treatment of brucellosis. According to the latest findings, the most effective immunogenic antigens of brucella to induce immune responses are included Omp31, BP26, BLS, DnaK and L7-L12. Therefore, in the present study, an in silico approach was used to design a novel multi-epitope vaccine to elicit a desirable immune response against brucellosis. First, five novel T-cell epitopes were selected from Omp31, BP26, BLS, DnaK and L7-L12 proteins using different servers. In addition, helper epitopes selected from Tetanus toxin fragment C (TTFrC) were applied to induce CD4+ helper T lymphocytes (HTLs) responses. Selected epitopes were fused together by GPGPG linkers to facilitate the immune processing and epitope presentation. Moreover, cholera toxin B (CTB) was linked to N terminal of vaccine construct as an adjuvant by using EAAAK linker. A multi-epitope vaccine was designed based on predicted epitopes which was 377 amino acid residues in length. Then, the physico-chemical properties, secondary and tertiary structures, stability, intrinsic protein disorder, solubility and allergenicity of this multi-epitope vaccine were assessed using immunoinformatics tools and servers. Based on obtained results, a soluble, and non-allergic protein with 40.59kDa molecular weight was constructed. Expasy ProtParam classified this chimeric protein as a stable protein and also 89.8% residues of constructed vaccine were located in favored regions of the Ramachandran plot. Furthermore, this multi-epitope peptide vaccine was able to strongly induce T cell and B-cell mediated immune responses. In conclusion, immunoinformatics analysis indicated that this multi-epitope peptide vaccine can be effectively expressed and potentially be used for prophylactic or therapeutic usages against brucellosis.

Keywords: BLS; BP26; Brucellosis; Dnak; L7–L12; Multi-epitope vaccine; Omp31.

MeSH terms

  • Acetyltransferases / genetics
  • Acetyltransferases / immunology
  • Amino Acid Sequence
  • Animals
  • Antigens, Bacterial / chemistry*
  • Antigens, Bacterial / genetics
  • Antigens, Bacterial / immunology
  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Outer Membrane Proteins / immunology
  • Bacterial Vaccines / biosynthesis*
  • Bacterial Vaccines / genetics
  • Bacterial Vaccines / pharmacology
  • Brucella / chemistry
  • Brucella / genetics
  • Brucella / immunology*
  • Brucellosis / immunology
  • Brucellosis / prevention & control*
  • Cholera Toxin / genetics
  • Cholera Toxin / immunology
  • Computational Biology
  • Epitopes, T-Lymphocyte / chemistry
  • Epitopes, T-Lymphocyte / genetics
  • Epitopes, T-Lymphocyte / immunology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • HSP70 Heat-Shock Proteins / genetics
  • HSP70 Heat-Shock Proteins / immunology
  • Humans
  • Membrane Proteins / genetics
  • Membrane Proteins / immunology
  • Models, Molecular
  • Protein Conformation, alpha-Helical
  • Protein Conformation, beta-Strand
  • Recombinant Fusion Proteins / chemistry*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Ribosomal Proteins / genetics
  • Ribosomal Proteins / immunology
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • T-Lymphocytes / cytology
  • T-Lymphocytes / drug effects
  • T-Lymphocytes / immunology
  • Vaccines, Subunit

Substances

  • Antigens, Bacterial
  • BP26 protein, Brucella
  • Bacterial Outer Membrane Proteins
  • Bacterial Vaccines
  • Epitopes, T-Lymphocyte
  • HSP70 Heat-Shock Proteins
  • Membrane Proteins
  • Omp31 protein, Brucella melitensis
  • Recombinant Fusion Proteins
  • Ribosomal Proteins
  • Vaccines, Subunit
  • Cholera Toxin
  • Acetyltransferases