Variation in expression of Biomphalaria glabrata SOD1: a potential controlling factor in susceptibility/resistance to Schistosoma mansoni.

The snail Biomphalaria glabrata kills the blood fluke Schistosoma mansoni by a mechanism involving production of hydrogen peroxide, the enzymatic product of cytosolic Cu/Zn superoxide dismutase (SOD1). This enzyme exhibits higher activity in blood cells (hemocytes) from a predominantly resistant strain of B. glabrata than in hemocytes from a susceptible strain. Additionally, B. glabrata SOD1 polymorphisms have been associated with susceptibility/resistance to the parasite. To address the hypothesis that SOD1 transcription levels differ in accordance with variation at the SOD1 locus, quantitative PCR was performed using hemocyte-derived cDNA prepared from SOD1-genotyped snails. Here we report that individuals possessing the allele previously associated with resistance to S. mansoni express significantly higher levels of hemocyte SOD1 transcripts than individuals lacking this allele. A causal relationship between SOD1 expression and susceptibility/resistance to S. mansoni is supported by the correlation of transcript quantity with data (from a previous study) on the probability of infection.

Biomphalaria glabrata cytosolic copper/zinc superoxide dismutase (SOD1) gene: association of SOD1 alleles with resistance/susceptibility to Schistosoma mansoni.

Variation in susceptibility of the snail Biomphalaria glabrata to infection by the parasite Schistosoma mansoni is, at least in part, genetically determined. Functional studies have demonstrated that hemocyte-mediated killing of the parasite involves hydrogen peroxide, the enzymatic product of superoxide dismutase (SOD). The present study identifies alleles of the gene coding for cytosolic copper/zinc SOD (SOD1). The resistance/susceptibility phenotypes and SOD1 genotypes were determined for 354 snails of the predominantly resistant 13-16-R1 strain of B. glabrata. Resistance to the parasite was found to be significantly associated with one allele of the SOD1 gene. Conversely, a separate SOD1 allele was significantly associated with susceptibility.

Respiratory burst of Biomphalaria glabrata hemocytes: Schistosoma mansoni-resistant snails produce more extracellular H2O2 than susceptible snails.

The production of reactive oxygen species by hemocytes from the gastropod Biomphalaria glabrata has been linked to their ability to kill the trematode parasite Schistosoma mansoni. For 2 laboratory strains of the snail, 1 resistant (13-16-R1) and 1 susceptible (MO) to the PR1 strain of S. mansoni, we compared hemocyte production of extracellular hydrogen peroxide when stimulated with the protein kinase C agonist phorbol myristate acetate (PMA). The time course of the PMA-induced response is similar in both strains with respect to onset, peak production, and termination of the respiratory burst. However, the magnitude of the response differs between strains, in that hemocytes from resistant snails generate significantly more hydrogen peroxide. These findings suggest that the capacity to produce hydrogen peroxide could be critical in determining susceptibility or resistance to S. mansoni.

Constitutive differences in Cu/Zn superoxide dismutase mRNA levels and activity in hemocytes of Biomphalaria glabrata (Mollusca) that are either susceptible or resistant to Schistosoma mansoni (Trematoda).

Genetic strains of the snail Biomphalaria glabrata vary in their resistance to the parasite Schistosoma mansoni. Phagocytic cells (hemocytes) circulating in the hemolymph of B. glabrata play an essential role in the snail's innate immune response. Hemocytes of resistant B. glabrata kill S. mansoni in vitro via a mechanism which involves a respiratory burst. Reactive oxygen species (ROS), which are products of the respiratory burst, can act as mediators of both oxidative damage and of immune-related intracellular signaling pathways. One specific ROS, hydrogen peroxide (H2O2), has been shown to be involved in hemocyte-mediated sporocyst killing. We tested the hypothesis that Cu/Zn superoxide dismutase (SOD), a cytosolic enzyme that catalyzes the conversion of superoxide anion to H2O2, is somehow different between resistant and susceptible snail strains. We report a hemocyte transcript with all the features of a typical cytosolic Cu/Zn SOD (GenBank accession numbers AY505496 and AY505497). The amount of Cu/Zn SOD mRNA in hemocytes from resistant snails was double that of hemocytes from susceptible snails, and this correlated directly with an increased Cu/Zn SOD enzymatic activity in resistant hemocytes. Additional experiments determined that in vitro interaction/encapsulation of sporocysts did not influence Cu/Zn SOD mRNA levels in hemocytes from either snail strain. Thus, resistance in this host-parasite system does not appear to depend on a transcriptional response of hemocyte Cu/Zn SOD, but may be due, at least in part, to a constitutively elevated enzymatic level of Cu/Zn SOD.

Schistosoma mansoni sporocysts in culture: host plasma hemoglobin contributes to in vitro oxidative stress.

The initiation and promotion of sporocyst propagation and subsequent production of cercariae by intramolluscan larval stages of digenic trematodes are thought to depend on mollusc-derived factors. The ability to investigate this using in vitro cultures of Schistosoma mansoni sporocysts has been impeded by the fact that plasma from the host, Biomphalaria glabrata, becomes toxic to the parasite in long-term cultures. The present study identifies hemoglobin as the plasma component responsible for this toxicity. The addition of the enzyme catalase to sporocyst cultures neutralized the toxic effects of both purified hemoglobin and whole plasma, suggesting that the generation of H2O2 as a consequence of hemoglobin oxidation is the mechanism of plasma toxicity. Furthermore, cultures incubated in unconditioned schistosome medium with plasma plus catalase yielded significantly higher numbers of daughter sporocysts than cultures with media or plasma alone, but not higher than cultures with catalase alone. These latter results suggest that the oxidative environment and the antioxidant capacity of the media are critical factors for in vitro propagation of S. mansoni sporocysts.