Synthetic Peptide Vaccine Development: Designing Dual Epitopes Into a Single Pilin Peptide Immunogen Generates Antibody Cross-Reactivity Between Two Strains of Pseudomonas Aeruginosa

Chem Biol Drug Des. 2010 Oct;76(4):293-304. doi: 10.1111/j.1747-0285.2010.01021.x. Epub 2010 Aug 30.

Abstract

One of the main challenges of Pseudomonas aeruginosa vaccine development is the design of an antigen that elicits cross-reactive antibodies against multiple virulent strains. Using a rational design approach, we have developed a single 17-residue peptide immunogen that generates antibodies that target the receptor-binding domain of the type IV pilus of more than one strain of P. aeruginosa. Using the receptor-binding domain sequence, of native strain PAO as a template, we have systematically changed up to five residues in the PAO sequence of the peptide immunogen into that of the PAK sequence. We show by indirect and competitive ELISA that the mutant peptide immunogens elicit the development of polyclonal sera that is cross-reactive to both native strain PAO and PAK pilin. We further show that there are at least two separate antibody populations in the polyclonal sera that possess closely related epitopes but which are each strain specific. Moreover, part of the epitope for the PAO-specific antibodies consists of several residues outside the disulfide loop of the receptor-binding domain. This allows us to create two unique epitopes within the same receptor-binding domain sequence.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution
  • Antibodies, Bacterial / immunology*
  • Antibodies, Bacterial / metabolism
  • Carrier Proteins / chemistry
  • Carrier Proteins / immunology
  • Cross Reactions
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / chemistry
  • Epitopes / immunology*
  • Fimbriae Proteins / chemistry
  • Fimbriae Proteins / genetics
  • Fimbriae Proteins / immunology*
  • Molecular Sequence Data
  • Mutation
  • Peptides / chemical synthesis
  • Peptides / chemistry
  • Peptides / immunology
  • Protein Binding
  • Protein Structure, Tertiary
  • Pseudomonas Vaccines / chemistry
  • Pseudomonas Vaccines / immunology*
  • Pseudomonas aeruginosa / immunology*
  • Vaccines, Synthetic / chemistry
  • Vaccines, Synthetic / immunology

Substances

  • Antibodies, Bacterial
  • Carrier Proteins
  • Epitopes
  • Peptides
  • Pseudomonas Vaccines
  • Vaccines, Synthetic
  • Fimbriae Proteins