title: Vibrio cholerae O139 conjugate vaccines: synthesis and immunogenicity of V. cholerae O139 capsular polysaccharide conjugates with recombinant diphtheria toxin mutant in mice.
abstract: Epidemiologic and experimental data provide evidence that a critical level of serum immunoglobulin G (IgG) antibodies to the surface polysaccharide of Vibrio cholerae O1 (lipopolysaccharide) and of Vibrio cholerae O139 (capsular polysaccharide [CPS]) is associated with immunity to the homologous pathogen. The immunogenicity of polysaccharides, especially in infants, may be enhanced by their covalent attachment to proteins (conjugates). Two synthetic schemes, involving 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) as activating agents, were adapted to prepare four conjugates of V. cholerae O139 CPS with the recombinant diphtheria toxin mutant, CRMH21G. Adipic acid dihydrazide was used as a linker. When injected subcutaneously into young outbred mice by a clinically relevant dose and schedule, these conjugates elicited serum CPS antibodies of the IgG and IgM classes with vibriocidal activity to strains of capsulated V. cholerae O139. Treatment of these sera with 2-mercaptoethanol (2-ME) reduced, but did not eliminate, their vibriocidal activity. These results indicate that the conjugates elicited IgG with vibriocidal activity. Conjugates also elicited high levels of serum diphtheria toxin IgG. Convalescent sera from 20 cholera patients infected with V. cholerae O139 had vibriocidal titers ranging from 100 to 3,200: absorption with the CPS reduced the vibriocidal titer of all sera to < or =50. Treatment with 2-ME reduced the titers of 17 of 20 patients to < or =50. These data show that, like infection with V. cholerae O1, infection with V. cholerae O139 induces vibriocidal antibodies specific to the surface polysaccharide of this bacterium (CPS) that are mostly of IgM class. Based on these data, clinical trials with the V. cholerae O139 CPS conjugates with recombinant diphtheria toxin are planned.
authors.affiliation: National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892-2720, USA. kossaczz@mail.nih.gov
authors.forename: Z
authors.initials: Z
authors.lastname: Kossaczka
authors.forename: J
authors.initials: J
authors.lastname: Shiloach
authors.forename: V
authors.initials: V
authors.lastname: Johnson
authors.forename: D N
authors.initials: DN
authors.lastname: Taylor
authors.forename: R A
authors.initials: RA
authors.lastname: Finkelstein
authors.forename: J B
authors.initials: JB
authors.lastname: Robbins
authors.forename: S C
authors.initials: SC
authors.lastname: Szu
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001770:Blood Bactericidal Activity; D022121:Cholera Vaccines / Q000276:immunology*; D004167:Diphtheria Toxin / Q000276:immunology*; D005260:Female; D007074:Immunoglobulin G / Q000097:blood; D051379:Mice; D011135:Polysaccharides, Bacterial / Q000276:immunology*; D018074:Vaccines, Conjugate / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D022121:Cholera Vaccines; D004167:Diphtheria Toxin; D007074:Immunoglobulin G; D011135:Polysaccharides, Bacterial; D018074:Vaccines, Conjugate; D014614:Vaccines, Synthetic