Immune status assessment by abundance of IFN-alpha and IFN-gamma mRNA in chicken blood

Avian diseases, including such viral infection as infectious bursal disease , infectious anemia, and Marek's disease, often cause immunosuppression, le ading to more severe infection, problems with secondary infection, and inad equate responses to vaccination. Immunosuppression thus causes serious econ omic losses in commercial poultry production. To date, methods for assessin g immune status have been too slow to be of practical help. Reasoning that immunosuppression should be reflected by reduced production of interferons (IFN) in response to a viral antigen, we have developed competitive nucleic acid hybridization microtiter plate assays for chicken IFN-alpha (ChIFN-al pha) and ChIFN-gamma mRNA. To evaluate the assay, chickens were challenged with inactivated Newcastle disease virus (iNDV). Whole blood samples were c ollected at various times subsequently and preserved with a cationic deterg ent. Later, total RNA was extracted, and mRNA for both ChIFN-alpha and ChIF N-gamma, was measured. Both rose from undetectable levels to reach a peak b y 4 h, remained high for about 3 days, and fell to undetectable levels by d ay 5. Results were similar in chickens aged between 1 and 28 days. In later experiments, blood was collected 4 h after viral challenge. When chickens were immunosuppressed by administering 4-5 mg cyclophosphamide (CY) daily f or 3 days and challenged with iNDV, they transcribed less ChIFN-alpha and C hIFN-gamma mRNA, and their antibody response was impaired. Our results sugg est that suspected immunosuppression in a commercial flock could be assesse d within 2-3 days by challenging birds with iNDV and measuring the amounts of ChIFN-alpha and ChIFN-gamma mRNA in blood obtained 2-4 h later.