Antiparasite activity of sea-anemone cytolysins on Giardia duodenalis and specific targeting with anti-Giardia antibodies

The killing activity of sea-anemone cytolysins on Giardia duodenalis was investigated. Three different toxins, sticho lysin I and II from Stichodactyla helianthus (St I and St II) and equinatoxin II from Actinia equina (EqtII) were all found to be active in an acute test, with a C-50 in the nanomolar range (St I, 0.5 nM; St II, 1.6 nM; and EqtII, 0.8 nM). A method to target the cytolysin activity more specifically towards the parasite cells by using anti-Giardia antibodies was then investigated. Parasite cells were sensitised with a primary murine monoclonal or polyclonal antibody followed by a biotinylated secondary anti-mouse-IgG monoclonal antibody. Subsequently, avidin and a biotinylated EqtII mutant were added, either in two separate steps or as a pre-formed conjugate. When the monoclonal antibody was used, the C-50 of biotinylated EqtII was 1.3 nM with sensitised cells and 5 nM with non-sensitised cells, indicating a four-fold enhancement of activity with the eel treatment. Treatment with the polyclonal antibody was somehow more effective than with the monoclonal antibody in an acute test. This indicates that sea-anemone cytolysins can efficiently kill Giardia cells, and that it is possible to improve, to a certain extent, the anti-parasite specificity of these toxins with anti-Giardia antibodies. However, the feasibility of this approach "in vivo" remains to be demonstrated. (C) 1999 Australian Society for Parasitology. Published by Elsevier Science Ltd. All rights reserved.