Opsonization of Streptococcus agalactiae of bovine origin by complement and antibodies against group B polysaccharide

Abstract

The contribution of bovine complement and antibodies (Ab) against the group B polysaccharidic antigen (GBA) to the opsonization of Streptococcus agalactiae isolated from bovine mastitis cases was investigated by using affinity-purified Ab. GBA-specific Ab were not opsonic by themselves, but in the presence of complement (precolostral calf serum) with an opsonization time of 15 min, they exhibited a dose-dependent opsonic activity in a polymorphonuclear leukocyte chemiluminescence assay. Kinetic studies of the deposition of complement component C3 on protein X-bearing nontypeable (NT/X) strains with an enzyme-linked immunosorbent assay showed that C3 was deposited on bacteria in the absence of Ab but that GBA-specific Ab markedly accelerated the process by reducing the lag phase, which extended up to 15 min when Ab were absent. In the absence of Ab, C3 deposition was inhibited by 5 mM salicylaldoxime or heat treatment at 56 degrees C for 3 min and necessitated Mg2+ ions but not Ca2+ ions, suggesting that activation of complement was effected by the alternative pathway only. When GBA-specific Ab were added to complement, the inhibitory treatments lost much of their efficacy, suggesting that the classical pathway was recruited. Deposition of C3 on NT/X strains in the absence of Ab induced chemiluminescence and phagocytic killing. With the addition of GBA-specific Ab, the numbers of surviving bacteria were halved (P < 0.05) compared with killing in the presence of complement alone. It can be concluded that NT/X strains are activators of the alternative pathway of complement and that GBA-specific Ab reinforce the opsonic efficiency of serum by recruiting the classical pathway and slightly enhancing phagocytic killing.