The production of toxic oxygen metabolites by hemocytes of different snail species.

The phagocytic hemocytes of four snail species were investigated for their ability to generate reactive oxygen metabolites upon stimulation by foreign material. Hemocytes of the pond snail Lymnaea stagnalis and of the garden snail Helix aspersa showed a luminol-dependent chemiluminescence (CL) when they phagocytosed zymosan particles. This CL was inhibited by superoxide dismutase (SOD) and sodium azide, indicating the involvement of oxygen intermediates. Hemocytes of the planorbid snails Planorbarius corneus and Biomphalaria glabrata did not give a detectable CL response. This is probably due to the presence of hemoglobin in the hemolymph; after isolation of the cells and subsequent stimulation, however, still no CL could be measured. Hemocytes of all four snail species showed a SOD-sensitive nitroblue tetrazolium (NBT) reduction, indicating the generation of superoxide anions. Regarding the NBT reaction, no differences were observed between strains of B. glabrata that were susceptible or resistant to PR-1 Schistosoma mansoni; neither did exposure to the parasite have an effect on the ability of the hemocytes to reduce NBT. Also, hemocytes from Trichobilharzia ocellata-infected L. stagnalis did not react differently from hemocytes of uninfected snails. It is now clear that phagocytically stimulated hemocytes of several molluscan species can generate reactive forms of oxygen; the relevance of this fact for the phylogeny of killing systems operative in leukocytes is discussed.