Wild birds, as reservoirs of viruses, play active roles in influenza virus transmission and reassortment. To further evaluate the potential infection risk of H6 AIVs from wild birds, we inoculated these viruses into A549 cells, which showed that these wild bird H6 AIVs have acquired the ability to cross interspecific barriers and have the potential to infect mammals. We analysed the level of cytokine expression in response to four H6 AIVs in CEFs and DEFs using real-time PCR. The results showed that the viruses from wild birds could induce a higher level of cytokine expression in CEFs than in DEFs. These findings suggested that H6 AIV in wild birds, especially migratory species, play an indispensable role in the spread and reassortment of AIVs and provide unknown opportunities for mutation and emergence of novel influenza viruses. Additional studies of AIVs originating from wild birds will help to determine viral adaptation and maintenance in alternative hosts.