ISOLATION AND FUNCTIONAL-CHARACTERIZATION OF SNAIL HEMOCYTE-MODULATING POLYPEPTIDE FROM PRIMARY SPOROCYSTS OF SCHISTOSOMA-MANSONI

Infection with larval trematodes has been shown to inhibit several snail-host defences, including hemocyte phagocytosis, cytotoxicity, motility, and adherence. Certain plasma factors which mediate snail defence responses, and which may be produced by host hemocytes, also appear to be altered by these parasites. In this study we present protocols for the isolation of 2 proteins from larval Schistosoma mansoni excretory-secretory (ES) products and detail the effects of these components on Biomphalaria glabrata hemocyte protein synthetic/secretory (S/S) activity. Schistosome ES proteins, separated with a combination of membrane ultrafiltration, size exclusion, and ion exchange chromatography, were tested for their in vitro effect on cultured snail hemocytes, in the presence and absence of homologous plasma. A high-molecular-weight ultrafiltration fraction of parasite ES products (H30), in combination with plasma, was found to differentially affect susceptible (M-line) and resistant (10-R2) snail hemocytes. Secretion of metabolically labeled polypeptides by M-line cells was inhibited significantly while the S/S response of 10-R2 hemocyte polypeptides was not affected. In the absence of homologous plasma, little or no differential affect of ES polypeptides on hemocyte S/S activity was seen. Much of the inhibitory activity of H30 was attributable to a-partially purified fraction, Peak I (PkI), of ES products. Evidence suggests that, in its native state, PkI is a high-molecular-weight protein aggregate comprising subunits of approximately 22-24 kDa. Thus, PkI, in the presence of homologous plasma components, is a potential mediator of schistosome-induced suppression of polypeptide synthesis or secretion in hemocytes of susceptible snails. In combination with other parasite and host factors, PkI may be involved in the host-parasite interaction which leads to the state of susceptibility or resistance found in our strains of B. glabrata.