α-GlcNAc-1→2-α-Glc, the Salmonella Homologue of a Conserved Lipopolysaccharide Motif in the Enterobacteriaceae, Elicits Broadly Cross-Reactive Antibodies

To define cross-reactive epitopes in Salmonella lipopolysaccharide (LPS), antisera designated anti-S, anti-Ra, and anti-Re were generated against smooth (S), complete-core (Ra), and deep-core mutant (Re) strains, respectively, and characterized immunochemically. The reactivities of anti-Ra and anti-S with rough LPS (rLPS) chemotypes in enzyme-linked immunosorbent assays (ELISA) decreased progressively with increasing truncation of the complete-core oligosaccharide (e.g., Ra > Rb(1) >…Re), while that of anti-Re increased (Ra < Rb(1) <…Re). Anti-Ra was relatively more reactive with nonhomologous smooth LPS (sLPS) than anti-S, which in turn was more reactive than anti-Re. This order reflected the relative reactivities of these sera with outer-core rLPS but not those with inner-core rLPS, which suggests that the cross-reactivities of all three sera with sLPS were mediated by antibodies which bind outer-core determinants. Anti-Ra, but not anti-S or anti-Re, reacted with molecules substituted by O chains in immunoblots and revealed ladder-like patterns in sLPSs of various serospecificities. Anti-Ra, however, did not react with O-antigen-specific neoglycoconjugates in ELISA, thus demonstrating specificity for core epitopes. Ra and Rb(1) but not other Salmonella core chemotypes inhibited the reactivity of anti-Ra with sLPS in ELISA, which showed that the terminal outer-core disaccharide, α-GlcNAc-1→2-α-Glc (GlcNAc→Glc), was the major epitope of cross-reactive antibodies in the serum. GlcNAc→Glc represents the conserved motif α-hexose-1→2-α-hexose in cores of the Enterobacteriaceae, other homologues of which should likewise be cross-reactive. These results demonstrate that S or Re strains do not elicit cross-reactive antibodies and indicate that immunization with Ra strains may represent a general strategy for eliciting cross-reactive antibodies against LPSs from enteric bacteria.