Intranasal Administration of Recombinant Influenza Vaccines in Chimeric Mouse Models to Study Mucosal Immunity

The overall goal of this procedure is to describe a system to evaluate the physiological initiation of vaccine specific adaptive immunity. This is accomplished first, using reverse genetics to generate live, attenuated influenza vaccines containing a specific immunogenic peptide, and then using a chick embryo to multiply the virus. Next cells are harvested from the lymph nodes of vaccinated mice and fax analysis is used to track dendritic cell migration.

In the next assay, CD eight positive T cells reactive to the immunogenic peptide are labeled and injected into vaccinated mice, followed by fax analysis to measure the proliferation of those cells to analyze the immune response in more detail. A competitive bone marrow chime mirror is made by infusing diptheria toxin receptor expressing bone marrow cells into irradiated congenic mice, which are then exposed to diptheria toxin. After a successful depletion, the mouse is vaccinated and labeled antibodies and tetramers are used to identify CD eight positive T cells.