Preparation of Anti-Aspergillus fumigatus Antibodies in egg laying hens and their protective efficacy in Cyclosporine A treated BALB-c mice - by Kohar Annie Berge Kissoyan.

Abstract

Background: Morbidity and mortality rates of Aspergillus fumigatus induced diseases in the immunocompromised population are on the rise. Limited availability of anti- fungal therapeutic agents and the emergence of resistant strains highlight the need of advancements in the field. While human vaccines for A. fumigatus are not available yet, promising results in animal models have been reported.Aim: Earlier we reported on the protective and therapeutic effects of hen-produced anti-A. fumigatus IgY antibodies in BALB-c mice. The present study dealt with its effect in cyclosporine A treated-BALB-c mice.Methods: Two egg-laying hens were injected subcutaneously (sc) with heat-killed A. fumigatus emulsified with Freund’s complete adjuvant, followed 12 days later with a booster shot of the heat-killed A. fumigatus emulsified with Freund’s incomplete adjuvant. Eggs laid were collected before and after immunization. IgY was extracted from the yolk, pooled, dialyzed and lyophilized. The presence of IgY anti-A. fumigatus antibodies in the post-immunization preparation was confirmed by ELISA and Polyacrylamide Gel Electrophoresis (PAGE).Daily intraperitoneal (ip) injections of cyclosporine A were administered to BALB-c mice for a period of 10 days. One group of cyclosporine treated-mice was given an ip injection of post- secondary immunization IgY 24 hours prior to, another group was given the injection simultaneously with, and a third group 24 hours after, challenge with an LD50 of A. fumigatus. A control cyclosporine A treated group was given post-primary immunization IgY. Other control groups included mice treated with cyclosporine A or diluent. Mice were monitored daily and the number of deaths in each group was recorded. To confirm that deaths were due to the fungus dead mice were dissected, lungs and kidneys were surgically removed and homogenates were cultured on Sabouraud Dextrose Agar.Results: All the mice died in the groups treated with cyclosporine A or cyclosporine A and post-primary-immunized IgY. Fi