Synovial fluid inhibits killing of Staphylococcus aureus by neutrophils

Serum in the extracellular environment promotes neutrophil bactericidal activity apart from its opsonizing properties. We examined the effect of non-inflammatory osteoarthritic synovial fluid on serum-mediated neutrophil killing of Staphylococcus aureus. This was done to evaluate the effect of synovial fluid on neutrophil bactericidal activity independent of opsonin concentration. With an initial inoculum of 5 X 10(6) CFU/ml, 1.47 +/- 0.14% bacteria survived after 120 min of incubation with 10% serum and neutrophils. In contrast, 4.07 +/- 0.33% bacteria survived after incubation in serum plus synovial fluid (P less than 0.001). This inhibitory effect was directly related to the concentration of synovial fluid in the incubation mixture. Increasing the concentration of synovial fluid resulted in an increased percent survival. Studies utilizing preopsonized bacteria and radiolabeled organisms demonstrated that synovial fluid did not interfere with opsonization or phagocytosis. Intracellular bactericidal activity was assayed separately from phagocytosis by utilizing a brief ingestion period followed by the removal of extracellular bacteria by either differential centrifugation or lysostaphin treatment. The reincubation of cells and associated bacteria with serum or serum plus synovial fluid revealed that synovial fluid significantly inhibited the promoting effect of serum on neutrophil bactericidal activity. After 60 min of incubation with 10% serum, 13.0 +/- 1.2% bacteria survived, whereas 21.5 +/- 2.3% survived after incubation in serum plus synovial fluid (P less than 0.005). Superoxide production was not affected by the presence of synovial fluid. These findings suggest that the inhibitory effect of synovial fluid is due to an interaction between synovial fluid and the serum factors that promote intracellular killing.