The use of synthetic peptides in the design of a consensus sequence vaccine for Pseudomonas aeruginosa.
Pseudomonas aeruginosa employs pili to mediate adherence to epithelial cell surfaces. Research has shown that the C-terminal region of the pilin monomer contains the epithelial cell binding domain, which is semiconserved in seven different strains of this bacterium. Antibodies to this region of the pilin molecule are also able to block and prevent the infection process. As there is a degree of sequence and structural homology in the C-terminal region and all strains examined have been shown to bind to the same cell surface receptor, we reasoned that it should be possible to produce a synthetic peptide consensus sequence which would provide cross-reactive antiserum from a single peptide immunogen inhibiting the adherence of the known strains of P. aeruginosa. In this article we examine the cross-reactivity of five rabbit polyclonal antisera. One has been raised against the cell-surface receptor binding domain of native PAK strain pilin (residues 128-144) while the others have been raised to analogues of this region. Analysis of the cross-reactivity of these antisera, using competitive ELISA assay, has shown that it is possible to manipulate the amino acid sequence of a peptide immunogen to generate antiserum, which exhibits enhanced cross-reactivity to various strains of P. aeruginosa. Furthermore, when this peptide is conjugated to tetanus toxoid and used to vaccinate mice it provided cross-reactive protection against heterologous challenge with PAO strain bacteria. The results of these experiments are analyzed, and the applicability of our hypothesis and the implications of this approach to the design of a strain-independent consensus vaccine for immunization against Pseudomonas aeruginosa are discussed.
Cachia
P J
PJ
Department of Biochemistry, University of Alberta, Canadian Bacterial Diseases Network of Centres of Excellence, Edmonton.
Glasier
L M
LM
Hodgins
R R
RR
Wong
W Y
WY
Irvin
R T
RT
Hodges
R S
RS
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000737:chemistry; D000942:Antigens, Bacterial / Q000737:chemistry; D001428:Bacterial Vaccines / Q000737:chemistry*; D015195:Drug Design; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D011552:Pseudomonas Infections / Q000517:prevention & control*; D011550:Pseudomonas aeruginosa; D011817:Rabbits
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001428:Bacterial Vaccines; D010455:Peptides