title: A Vibrio cholerae classical TcpA amino acid sequence induces protective antibody that binds an area hypothesized to be important for toxin-coregulated pilus structure.
abstract: Vibrio cholerae is a gram-negative bacterium that has been associated with cholera pandemics since the early 1800s. Whole-cell, killed, and live-attenuated oral cholera vaccines are in use. We and others have focused on the development of a subunit cholera vaccine that features standardized epitopes from various V. cholerae macromolecules that are known to induce protective antibody responses. TcpA protein is assembled into toxin-coregulated pilus (TCP), a type IVb pilus required for V. cholerae colonization, and thus is a strong candidate for a cholera subunit vaccine. Polypeptides (24 to 26 amino acids) in TcpA that can induce protective antibody responses have been reported, but further characterization of their amino acid targets relative to tertiary or quaternary TCP structures has not been done. We report a refinement of the TcpA sequences that can induce protective antibody. One sequence, TcpA 15 (residues 170 to 183), induces antibodies that bind linear TcpA in a Western blot as well as weakly bind soluble TcpA in solution. These antibodies bind assembled pili at high density and provide 80 to 100% protection in the infant mouse protection assay. This is in sharp contrast to other anti-TcpA peptide sera (TcpA 11, TcpA 13, and TcpA 17) that bind very strongly in Western blot and solution assays yet do not provide protection or effectively bind TCP, as evidenced by immunoelectron microscopy. The sequences of TcpA 15 that induce protective antibody were localized on a model of assembled TCP. These sequences are centered on a site that is predicted to be important for TCP structure.
authors.affiliation: Department of Microbiology and Immunology, Dartmouth Medical School, Lebanon, New Hampshire 03756, USA.
authors.forename: Ronald K
authors.initials: RK
authors.lastname: Taylor
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Kirn
authors.forename: Michael D
authors.initials: MD
authors.lastname: Meeks
authors.forename: Terri K
authors.initials: TK
authors.lastname: Wade
authors.forename: William F
authors.initials: WF
authors.lastname: Wade
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D000907:Antibodies, Bacterial / Q000276:immunology*; D000918:Antibody Specificity; D002771:Cholera / Q000276:immunology / Q000517:prevention & control; D022121:Cholera Vaccines / Q000737:chemistry / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D036781:Fimbriae Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D010861:Fimbriae, Bacterial / Q000737:chemistry* / Q000276:immunology*; D007106:Immune Sera / Q000276:immunology; D051379:Mice; D008958:Models, Molecular; D011487:Protein Conformation; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012995:Solubility; D015996:Survival Rate; D014734:Vibrio cholerae / Q000276:immunology* / Q000648:ultrastructure; D014774:Virulence
chemical_list: D000907:Antibodies, Bacterial; D022121:Cholera Vaccines; D007106:Immune Sera; D011994:Recombinant Proteins; C085807:TcpA protein, Vibrio cholerae; D036781:Fimbriae Proteins