Abstract: In order to evaluate the antibody dynamic in different immune routes and immune protection of the recombinant fowlpox virus coexpressing cytokine IL6 gene and HA gene of H5 subtype avian influenza virus (rFPVAIH5AIL6). Groups of 10dayold SPF chickens were inoculated by subcutaneous route in the back of neck and wings route with inoculums containing rFPVAIH5AIL6. There was no significant difference in body weight between the group of rFPVAIH5AIL6 and the normal control group, but the body weight in group of wild type strain was lower than that of the normal control group. The result of HI antibodies showed that the chickens inoculated subcutaneously by wing route produced higher level HI antibody. HI antibody induced by rFPVAIH5AIL6 was higher than that induced by rFPVAIH5A. HI antibody reached a peak at 21 days post vaccination，decreased at 28 days post vaccination，and remained at a certain level at 49 days post vaccination. Another Group of 10dayold SPF chickens was inoculated by subcutaneous route in wings with inoculums containing rFPVAIH5AIL6. The chickens were challenged with H5 subtype avian influenza virus at 21 days post vaccination. The protection indexes of groups immunized with rFPVAIH5AIL6 and killed vaccine were 95%, significantly higher than that of other groups. The result of virus shedding showed that the shedding rate of chickens immunized with rFPVAIH5AIL6 was the lowest. In summary, chicken IL6 could improve the immune efficacy against AI when it was coexpressed with HA gene in recombinant fowlpox virus. This study paves the way for further development of a new AIV recombinant vaccine.