Alternative complement pathway activation by Salmonella O polysaccharide as a virulence determinant in the mouse

The quality of Salmonella O polysaccharide (the O antigen) is a virulence factor in mouse salmonellosis. It affects the rate by which these bacteria are phagocytosed and by which they activate the alternative complement pathway in a manner inversely proportional to their virulence, suggesting that the rate of complement activation is crucial for the fate of the bacteria in the mouse. The effector mechanism has, however, remained open since Salmonellae survive and multiply in the macrophages of the mouse. We show in this study that although the least virulent O-6,7 Salmonellae multiply in the liver macrophages they are rapidly killed in the peritoneal cavity by the local resident macrophages. Electron microscopy showed a striking morphological feature--a 35 nm thick homogenous electron-dense deposit--on all the bacteria found in association with the macrophages but absent from all non-cell-associated bacteria. A similar precipitate was formed by incubating the bacteria in fresh mouse serum and was dependent on heat-labile serum components and bound anti-C3. The least virulent O-6,7 bacteria acquired this deposit more rapidly and in a lower concentration of serum than the more virulent O-4,12 bacteria consistent with the previously demonstrated difference between these bacteria in their rate of complement activation via the alternative pathway. Preincubation of the O-4,12 bacteria in fresh mouse serum leading to complement deposition on 80% of the bacteria effectively opsonized them for rapid killing in the peritoneal cavity. These data for the first time demonstrate how the rate of complement activation determines the virulence of Salmonellae.