Aim:To investigate a new strategy to enhance the efficacy of a recombinantpertussis DNA vaccine.The strategy is co-injection with cytokine plasmids asprime,and boosted with purified homologous proteins.Method:A recombinantpertussis DNA vaccine containing the pertussis toxin subunit 1(PTS 1),fragmentsof the filamentous hemagglutinin(FHA)gene and pertactin(PRN)gene encodingfilamentous hemagglutinin and pertactin were constructed.Balb/c mice were im-munized with several DNA vaccines and antigen-specific antibodies anti-PTS1,anti-PRN,anti-FHA,cytokines interleukin(IL)-10,IL-4,IFN-γ,TNF-α,and spleno-cyte-proliferation assay were used to describe immune responses.Results:Therecombinant DNA vaccine could elicit similar immune responses in mice as that ofseparate plasmids encoding the 3 fragments,respectively.Mice immunized withDNA and boosted with the corresponding protein elicited more antibodies thanthose that received DNA as boost.In particular,when the mice were co-immu-nized with murine granulocyte-macrophage colony-stimulating factor plasmidsand boosted with proteins,all 4 cytokines and the 3 antigen-specific antibodieswere significantly increased compared to the pVAX1 group.Anti-PTS1,anti-FHA,IL-4 and TNF-α elicited in the colony stimulating factor(CSF)prime-proteinboost group showed significant increase compared to all the other groups.Conclusion:This prime and boost strategy has proven to be very useful in im-proving the immunogenicity of DNA vaccines against pertussis. Aim: To investigate a new strategy to enhance the efficacy of a recombinantpertussis DNA vaccine. The strategy is co-injection with cytokine plasmids asprime, and boosted with purified homologous proteins. Method: A recombinantpertussis DNA vaccine containing the pertussis toxin subunit 1 (PTS 1 ) gene, pertactin (PRN) gene encoding filamentous hemagglutinin and pertactin were constructed. Balb / c mice were im-munized with several DNA vaccines and antigen-specific antibodies anti-PTS1, anti-PRN, anti- FHA, cytokines interleukin (IL) -10, IL-4, IFN-γ, TNF-α, and spleno-cyte-proliferation assay were used to describe immune responses. Results: Therecombinant DNA vaccine could elicit similar immune responses in mice as that ofseparate plasmids encoding the 3 fragments, respectively. Mice immunized with DNA and boosted with the corresponding protein elicited more antibodies thanthose that DNA as boost.In particular, when the mice were co-immu-nized with murin e granulocyte-macrophage colony-stimulating factor plasmids and boosted with proteins, all 4 cytokines and the 3 antigen-specific antibodies were significantly increased compared to the pVAX1 group. Anti-PTS1, anti-FHA, IL-4 and TNF-α elicited in the colony stimulating factor (CSF) prime-proteinboost group showed significant increase compared to all the other groups. Confluence: This prime and boost strategy has proven to be very useful in im-proving the immunogenicity of DNA vaccines against pertussis.