Pseudomonas aeruginosa outer-membrane protein F epitopes are highly immunogenic in mice when expressed on a plant virus

Microbiology (Reading). 1999 Jan:145 ( Pt 1):211-220. doi: 10.1099/13500872-145-1-211.

Abstract

A synthetic peptide (peptide 10) representing a surface-exposed, linear B cell epitope from outer-membrane (OM) protein F of Pseudomonas aeruginosa was shown previously to afford protection in mice from P. aeruginosa infection. This peptide was expressed in tandem with the protein F peptide 18 on each of the two coat proteins of cowpea mosaic virus (CPMV). The chimaeric virus particles (CVPs) expressing the peptides on the S (small) coat protein (CPMV-PAE4) and L (large) coat protein (CPMV-PAE5) were used to immunize mice. Following subcutaneous immunization in Freund's and QuilA adjuvants, CPMV-PAE4 induced antibodies predominantly against peptide 18, whereas CPMV-PAE5 produced antibodies exclusively against peptide 10, indicating that the site of peptide expression on CPMV influences its immune recognition. The anti-peptide antibodies elicited by CPMV-PAE5 were predominantly of the IgG2a isotype, indicating a highly polarized TH1-type response. The peptide-specific IgG2a strongly recognized the whole F protein, but more importantly, recognized protein F in all seven Fisher-Devlin immunotypes of P. aeruginosa. Furthermore, the peptide-specific IgG2a in CVP/QS-21 adjuvant-immunized mice was shown to bind complement and to augment phagocytosis of P. aeruginosa by human neutrophils in vitro. The ability of CPMV-PAE5 to induce P. aeruginosa-specific opsonic IgG2a gives it potential for further development as a protective vaccine against P. aeruginosa.

MeSH terms

  • Animals
  • Antibodies, Bacterial / biosynthesis*
  • Capsid / biosynthesis
  • Capsid / genetics
  • Comovirus / genetics*
  • Comovirus / immunology
  • Complement System Proteins / immunology
  • Epitopes / genetics
  • Epitopes / immunology*
  • Genetic Vectors / genetics
  • Humans
  • Immunization
  • Immunoglobulin G / biosynthesis
  • Immunoglobulin G / immunology
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Neutrophils / immunology
  • Opsonin Proteins / blood
  • Opsonin Proteins / immunology
  • Phagocytosis
  • Porins / genetics
  • Porins / immunology*
  • Pseudomonas Infections / classification
  • Pseudomonas Infections / immunology
  • Pseudomonas Infections / microbiology
  • Pseudomonas Infections / prevention & control
  • Pseudomonas aeruginosa / immunology*
  • Th1 Cells / immunology
  • Vaccines, Synthetic / administration & dosage
  • Vaccines, Synthetic / genetics
  • Vaccines, Synthetic / immunology

Substances

  • Antibodies, Bacterial
  • Epitopes
  • Immunoglobulin G
  • OmpF protein
  • Opsonin Proteins
  • Porins
  • Vaccines, Synthetic
  • Complement System Proteins