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

Int J Parasitol. 1999 Mar;29(3):489-98. doi: 10.1016/s0020-7519(98)00220-3.

Abstract

The killing activity of sea-anemone cytolysins on Giardia duodenalis was investigated. Three different toxins, sticholysin 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 C50 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 C50 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 cell 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / immunology
  • Antibodies, Protozoan / immunology*
  • Antibodies, Protozoan / metabolism
  • Avidin / metabolism
  • Biotin / metabolism
  • Biotinylation
  • Cnidarian Venoms / immunology*
  • Cnidarian Venoms / metabolism
  • Cnidarian Venoms / pharmacology
  • Cytotoxicity, Immunologic
  • Cytotoxins / immunology*
  • Cytotoxins / metabolism
  • Cytotoxins / pharmacology
  • Giardia lamblia / drug effects
  • Giardia lamblia / immunology*
  • Giardiasis / parasitology
  • Immunotoxins / immunology*
  • Immunotoxins / metabolism
  • Mice
  • Sea Anemones*

Substances

  • Antibodies, Monoclonal
  • Antibodies, Protozoan
  • Cnidarian Venoms
  • Cytotoxins
  • Immunotoxins
  • Avidin
  • equinatoxin
  • Biotin