CHARACTERIZATION OF THE CHEMILUMINESCENCE MEASURED IN HEMOCYTES OF THE EASTERN OYSTER, CRASSOSTREA-VIRGINICA

Hemocytes of the Eastern oysters, Crassostrea virginica, produce react ive oxygen intermediates (ROIs) during phagocytosis to destroy foreign cells. Although evidence suggests that oyster hemocytes generate supe roxide anions (Oz) following phagocytic stimulation, the production of other ROI species such as hydrogen peroxide (H2O2), singlet oxygen (O -1(2)), hydroxyl radical (. OH), and hypochlorous acid (HOCl) has not been directly investigated. In this study, ROI production by oyster he mocytes was measured by quantifying the production of chemiluminescenc e (CL) amplified with different luminescent probes. The ROIs involved in CL production were identified by testing the effects of scavengers and inhibitors of particular ROI species on CL. Taurine completely inh ibited luminol-dependent CL, indicating that HOCl production was prima rily responsible for the CL generated by hemocytes during phagocytosis of yeast granules. In addition, azide strongly inhibited luminol CL, implicating the involvement of myeloperoxidase in the production of HO Cl and the resulting CL. Superoxide dismutase partially inhibited CL, indicating that superoxide ions were also produced and contributed to CL. Luminol CL was significantly higher in hemocytes from oysters heav ily infected with the parasite Perkinsus marinrcs and remained complet ely inhibitable by taurine suggesting that HOCl production was enhance d by infection. Last, the concentration of taurine measured in oyster hemolymph was sufficient to quench a significant amount of the HOCl ge nerated by the hemocytes and may reduce the effectiveness of the oyste r's defense response to infections. (C) 1995 Wiley-Liss, Inc.