Cytotoxicity of Echinococcus granulosus cyst fluid in vitro.

In a cytotoxicity assay, using rat spleen cells as target cells, Echinococcus granulosus cyst fluid from cattle exerted a marked degree of cytotoxicity in vitro. When trypan blue exclusion or [3H]thymidine incorporation by concanavalin A stimulated spleen cells was used as a measure of cell viability, dialyzed cyst fluid showed maximum cell destruction up to a 1:8 dilution. The effect was dose and time dependent, cells being affected by 24 h after exposure to cyst fluid. The components responsible for cytotoxicity of cyst fluid were heat stable and could be recovered using gel chromatography on Sephadex G 50 and G 15 as a single low molecular weight fraction. It is assumed that the toxic products released by the living parasite can readily penetrate through the cyst wall into the surrounding host tissue. The interference of such substances with immunocompetent cells might account for the long-term survival of the parasite in the intermediate host.

[Simple method for obtaining Toxocara canis antigen for the indirect immunofluorescence technic]. / Eine einfache Methode zur Gewinnung von Toxocara canis-Antigen für die indirekte Immunofluoreszenz-Technik

A simple method for the preparation of Toxocara canis antigen for the indirect immunofluorescent test is described: Embryonated Toxocara eggs are treated for 12 hours at room temperature with a 1:1 mixture of 2% NaHO and sodium hypochlorite (NaClO) solution with a concentration of 2% free chlorine in order to remove the outer layers of the egg shells. The larvae which are still enclosed in the lipid membrane are freed by mild ultrasonic treatment. Thereafter, the suspension of larvae is washed and purified in a modified Baermann apparatus. In this way large numbers of larvae in pure suspension were gained and used for the production of frozen sections for the indirect immunofluorescent test. Rabbits and mice experimentally infected with embryonated Toxocara canis eggs showed a positive serological reaction (titers between 1/10 to 1/320) in this test with Toxocara larvae as antigen, while in uninfected control animals no antibodies could be detected. The larval antigen exhibited only a weak cross reaction with sera of animals infected with Ascaris suum eggs.