Naturally acquired antibodies against four Enterococcus faecalis capsular polysaccharides in healthy human sera

Abstract

Healthy human sera (HHS) contain naturally acquired enterococcal antibodies which promote neutrophil-mediated killing. The target antigens remain unknown. The present study used a capsular polysaccharide (CPS)-enzyme-linked immunosorbent assay (ELISA) to investigate whether the HHS antibodies of 12 healthy donors bound to the CPS of four E. faecalis serotypes (CPS-A to CPS-D) and then employed an opsonic-killing assay to determine if these antibodies mediated phagocyte-dependent killing. All HHS contained immunoglobulin G (IgG) and IgM antibodies directed against capsular polysaccharides of the four serotypes. Absorption of the sera with homologous and heterologous strains showed a majority of antibodies to be cross-reactive among the prototype strains. The susceptibility of the four prototype strains to opsonic killing varied. Opsonic killing of CPS-A and CPS-B strains was significantly higher than killing of CPS-C and CPS-D strains. Absorption studies revealed that the opsonic killing of HHS was only partially type specific, with cross-reactivity between CPS-A and CPS-B strains and between CPS-C and CPS-D strains. These data indicate that healthy individuals possess opsonic antibodies specific for CPS-A and CPS-B but only low titers of opsonic antibodies against CPS-C and CPS-D. Titers of opsonic antibodies did not correlate with antibody titers measured by ELISA. Whether this lack of correlation is due to the low frequency of opsonic antibodies or to increased resistance to the opsonophagocytic killing of some serotypes remains to be determined.

FIG. 1.

IgG (A) and IgM (B) concentrations (CPS-ELISA) against the capsular polysaccharides of four E. faecalis prototype strains in the sera of 12 healthy humans. Horizontal lines show the average antibody concentrations. The average concentrations for both anti-CPS IgG and anti-CPS IgM among the four capsular serotypes are not statistically different (P > 0.05).

FIG. 2.

Opsonophagocytic killing of six representative human sera (serum dilution of 1:1,000) of healthy volunteers against the four E. faecalis prototype strains. Horizontal lines show the average percent killing.