Integrin-like RGD-dependent binding mechanism involved in the spreading response of circulating molluscan phagocytes

Circulating phagocytic cells (hemocytes) of the snail Biomphalaria glabrata, intermediate host of the human blood fluke Schistosoma mansoni, were treated with the tetrapeptide, arg-gly-asp-ser (RGDS), an integrin-specific adhesion inhibitor, and assessed for their ability to adhere and spread on uncoated and snail plasma protein-coated glass slides. Although cells were capable of adherence, RGDS significantly inhibited the spreading ability of hemocytes in both a time and RGDS concentration-dependent fashion regardless of plasma protein coating. The inhibition of hemocyte spreading by RGDS was a specific response, since treatment of cells dth a glutamic acid-substituted control peptide (RGES) did not exert the same inhibitory effect. A comparison of RGDS-responses between hemocytes of two strains of B. glabrata, one resistant (R; 13-16-R1 strain) and the other susceptible (S; NMRI strain) to infection by S. mansoni, revealed several snail strain-specific differences. At concentrations of 0.5 mM RGDS, R snail hemocyte spreading was unaffected, whereas a significant depression of spreading was seen in cells of the S snail. Moreover, we observed that R strain hemocytes spread more rapidly on homologous plasma-coated surfaces than the S snail strain following peptide pretreatment and removal. These data suggest that hemocytes from S and R snails may differ either in the number of RGDS-binding receptors or in their affinity for the PODS peptide. In order to identify the type(s) of integrin-like POD-binding receptors that may be present on the surface of snail immunocytes, washed hemocytes were placed on various mammalian extracellular matrix proteins and evaluated for their spreading function in the presence of specific or non-specific peptides. Hemocyte aggregation or clumping was observed on all test protein substrates, and this aggregation behavior was specifically inhibited by PODS. Thus, RGD-binding receptors appear to play a critical role in cellular motility on matrix-coated surfaces and/or cell-cell binding. Our data provide functional evidence for an integrin-like receptor on circulating phagocytes of snails, and for an RGD-binding mechanism involved in cell-substrate interactions. (C) 1998 Published by Elsevier Science Ltd. All rights reserved.