title: Synthetic peptide vaccine development: designing dual epitopes into a single pilin peptide immunogen generates antibody cross-reactivity between two strains of Pseudomonas aeruginosa.
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.
authors.affiliation: Department of Biochemistry and Molecular Genetics, University of Colorado Denver, School of Medicine, Program in Structural Biology and Biophysics, 12801 E. 17th Ave, MS 8101, Aurora, CO 80045, USA.
authors.forename: Clifton
authors.initials: C
authors.lastname: Hackbarth
authors.forename: Robert S
authors.initials: RS
authors.lastname: Hodges
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000907:Antibodies, Bacterial / Q000276:immunology* / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D036781:Fimbriae Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D009154:Mutation; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D054406:Pseudomonas Vaccines / Q000737:chemistry / Q000276:immunology*; D011550:Pseudomonas aeruginosa / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000737:chemistry / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D002352:Carrier Proteins; D000939:Epitopes; D010455:Peptides; D054406:Pseudomonas Vaccines; D014614:Vaccines, Synthetic; D036781:Fimbriae Proteins
references.citation: Mol Microbiol. 1994 Feb;11(4):705-13
references.pmid: 7910938
references.citation: Biochemistry. 2001 Apr 3;40(13):3985-95
references.pmid: 11300779
references.citation: Biochemistry. 2004 Sep 14;43(36):11427-35
references.pmid: 15350129
references.citation: Infect Immun. 1991 Mar;59(3):822-8
references.pmid: 1671777
references.citation: J Pept Res. 1998 Oct;52(4):289-99
references.pmid: 9832307
references.citation: Nat Rev Microbiol. 2004 May;2(5):363-78
references.pmid: 15100690
references.citation: J Mol Biol. 2000 Jun 16;299(4):1005-17
references.pmid: 10843854
references.citation: Bioconjug Chem. 1990 Mar-Apr;1(2):114-22
references.pmid: 2095209
references.citation: Nature. 2000 Aug 31;406(6799):959-64
references.pmid: 10984043
references.citation: Biochemistry. 1995 Dec 19;34(50):16255-68
references.pmid: 8845350
references.citation: Adv Microb Physiol. 1988;29:53-114
references.pmid: 2898203
references.citation: Biopolymers. 2003;71(2):141-68
references.pmid: 12767116
references.citation: Biochemistry. 1993 Dec 14;32(49):13432-40
references.pmid: 8257679
references.citation: Infect Immun. 1995 Apr;63(4):1278-85
references.pmid: 7890385
references.citation: Annu Rev Microbiol. 2002;56:289-314
references.pmid: 12142488
references.citation: J Biol Chem. 1988 Aug 25;263(24):11768-75
references.pmid: 2457026
references.citation: J Mol Biol. 2007 Nov 23;374(2):426-42
references.pmid: 17936788
references.citation: J Biomol NMR. 2001 Apr;19(4):385-6
references.pmid: 11370789
references.citation: Methods Enzymol. 1995;253:115-31
references.pmid: 7476380
references.citation: Infect Immun. 1994 Oct;62(10):4118-23
references.pmid: 7927665
references.citation: J Biol Chem. 2001 Jun 29;276(26):24186-93
references.pmid: 11294863
references.citation: Clin Infect Dis. 2005 Sep 15;41(6):848-54
references.pmid: 16107985
references.citation: Protein Sci. 1992 Oct;1(10):1308-18
references.pmid: 1284654
references.citation: Mol Cell. 2003 May;11(5):1139-50
references.pmid: 12769840