Animal models are essential to examine the pathogenesis and transmission of influenza viruses and for preclinical evaluation of influenza virus vaccines. Among the animal models used in influenza virus research, the domestic ferret (Mustela putorius furo) is the gold standard. As seen in humans, infection with influenza virus or immunization with an influenza virus vaccine induces humoral and cellular immunity in ferrets that provides protection against infection by an antigenically similar influenza virus. Antibodies against the globular head domain of the influenza hemagglutinin can provide sterilizing immunity against virus infection by blocking receptor binding. However, antibodies that bind the stalk region of the hemagglutinin also confer protection by several mechanisms including antibody-dependent cellular cytotoxicity or phagocytosis. Recently, the antigenically and structurally conserved hemagglutinin stalk has become an attractive target for the development of universal influenza virus vaccines that hold the promise to provide protection against influenza epidemics and pandemics. Herein, in vivo and in vitro assays, including optimization of assay conditions to examine hemagglutinin stalk-specific antibody responses in small animal models, are described.
Keywords: ELISA; ELISpot; Ferret; Flow cytometry; HI assay; Hemagglutinin stalk immunity; Influenza virus.