In vitro and in vivo assay systems for study of influenza virus inhibitors.

Evaluation of potential influenza virus inhibitors may utilize multiple steps. First would be to determine if the viral target (e.g. influenza virus neuraminidase) being focused upon will be inhibited in the appropriate assay. Standard in vitro antiviral assays, used next in antiviral evaluations, may utilize inhibition of viral plaques, viral cytopathic effect (CPE), and viral hemagglutinin or other protein, with inhibition of viral yield used in follow-up evaluations. The CPE can be determined visually and by dye uptake. Animal models used for study of potential influenza virus inhibitors include the ferret, the laboratory mouse, and the chicken, with a variety of parameters used to indicate the severity of the infection and its inhibition by therapy. Multiple parameters are recommended in any in vivo antiviral evaluation. The ferret and the mouse infection models have been useful in studying the development of drug resistance and the relative virulence of drug-resistant viruses. The influenza mouse model has also been of value for the evaluation of immunomodulating effects of test compounds and for the study of the utility of antiviral drugs for use against influenza virus infections in the immunocompromised host. In considering the use of any animal model, species differences in drug pharmacology and metabolism must be taken into account. This review has described the systems which have been used most frequently by antiviral investigators, using, as examples, recent studies with the clinically approved influenza virus neuraminidase inhibitors oseltamivir and zanamivir.