Characterization of cytokine expression induced by avian influenza virus infection with real-time RT-PCR

Abstract

Knowledge of how birds react to infection from avian influenza virus is critical to understanding disease pathogenesis and host response. The use of real-time (R) RT-PCR to measure innate immunity, including cytokine and interferon gene expression, has become a standard technique employed by avian immunologists interested in examining these responses. This technique utilizes nucleotide primers and fluorescent reporter molecules to measure amplification of the gene of interest. The use of RRT-PCR negates the need for northern blot analysis or DNA sequencing. It is simple, specific and sensitive for the gene of interest. However, it is dependent on knowing the target sequence prior to testing so that the optimal primers can be designed. The recent publication of genomic sequences of Gallus gallus, Meleagris gallopavo, and Anas platyrhynchos species makes it possible to measure cytokine expression in chicken, turkey, and duck species, respectively. Although these tests do not measure functionally expressed protein, the lack of antibodies to identify and quantify avian cytokines from different avian species makes this technique critical to any characterization of innate immune responses through cytokine and interferon activation or repression.