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ABSTRACTThe epidemic and pandemic potential of Vibrio choleraeO139 is such that a vaccine against this newly emerged serogroup ofV. choleraeis required. A conjugate made of the polysaccharide moiety (O-specific polysaccharide plus core) of the lipopolysaccharide (LPS) of V. choleraeO139 (pmLPS) was prepared by derivatization of the pmLPS with adipic acid dihydrazide and coupling to tetanus toxoid (TT) by carbodiimide-mediated condensation. The immunologic properties of the conjugate were tested using BALB/c mice injected subcutaneously three times at 2 weeks interval and then a fourth time 4 weeks later. Mice were bled 7 days after each injection and then once each month for the following 6 months. LPS and TT antibody levels were determined by enzyme-linked immunosorbent assay using immunoplates coated with either O139 LPS or TT. Both pmLPS and pmLPS-TT conjugate elicited low levels of immunoglobulin M (IgM), peaking 5 weeks after the first immunization. The conjugate elicited high levels of IgG antibodies, peaking 3 months after the first immunization and declining slowly during the following 5 months. TT alone, or as a component of conjugate, induced mostly IgG antibodies. Antibodies elicited by the conjugate recognized both capsular polysaccharide and LPS from V. choleraeO139 and were vibriocidal. They were also protective in the neonatal mouse model of cholera infection. The conjugation of the O139 pmLPS, therefore, enhanced its immunogenicity and conferred T-dependent properties to this polysaccharide.

Staphylococcus aureusexpresses at least eight distinct serotypes of capsular polysaccharide (CP). Gene clusters involved in the expression of serotypes 1, 5 and 8 have been cloned and sequenced. In this report we describe the isolation and analysis of serotype 5 capsular polysaccharide‐defective mutants. A naturally occurring capmutation in the laboratory strains 8325‐4 and RN4220 was mapped to the cap5Egene by genetic complementation. The cap5H–Kgenes were shown to be responsible for CP5 serotype specificity by transduction and complementation.

Staphylococcus aureus expresses at least eight distinct serotypes of capsular polysaccharide (CP). Gene clusters involved in the expression of serotypes 1, 5 and 8 have been cloned and sequenced. In this report we describe the isolation and analysis of serotype 5 capsular polysaccharide-defective mutants. A naturally occurring cap mutation in the laboratory strains 8325-4 and RN4220 was mapped to the cap5E gene by genetic complementation. The cap5H–K genes were shown to be responsible for CP5 serotype specificity by transduction and complementation.