An enzyme-release assay for the assessment of the lytic activities of complement or antimicrobial peptides on extracellular Toxoplasma gondii.
A method is described which allows the evaluation of the membrane lytic activity of either complement or antimicrobial peptides against the extracellular stage of the human protozoan parasite Toxoplasma gondii. The assay is based on lacZ transgenic parasites, determining the activity of released cytoplasmic beta-galactosidase into the culture supernatant upon membrane disintegration. This method was used to evaluate the lytic activities of (i) complement which is a natural defense mechanism in infected hosts against extracellular parasites, and (ii) antimicrobial peptides which have not been evaluated against T. gondii before. The results show that the assay provides a simple and convenient way to assess the membrane lytic activity of such compounds and that T. gondii, like other protozoan parasites, is vulnerable to the membrane-lytic effect of antimicrobial peptides.
Seeber
F
F
FB Biologie/Parasitologie, Philipps-Universität Marburg, Germany. seeber@mailer.uni-marburg.de
D000595:Amino Acid Sequence; D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology; D003165:Complement System Proteins / Q000276:immunology / Q000494:pharmacology*; D006801:Humans; D008555:Melitten / Q000737:chemistry; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D014122:Toxoplasma / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics / Q000276:immunology; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism*
D000981:Antiprotozoal Agents; D010455:Peptides; D008555:Melitten; D003165:Complement System Proteins; D001616:beta-Galactosidase