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Porcine epidemic diarrhea virus (PEDV) is the causative agent of porcine epidemic diarrhea, a highly contagious enteric disease of swine.The Spike (S) protein is one of the main structural proteins of PEDV capable of inducing neutralizing antibodies in vivo.Herein, we generated three distinct DNA constructs in the eukaryotic expression plasmid pVAX1;one encoding the S protein [pVAX1-(PEDV-S)], the second encoding the N-terminal fragment (S1) [pVAX1-(PEDV-S1)] containing potent antigenic sites, and the third expressing the porcine interleukin-18 (pIL-18) [pVAX1-(IL-18)].Immunofluorescence assays in BHK-21 cells demonstrated successful protein expression from all 3 constructs.Kunming mice were injected separately with each of these construct or with a pVAX1-(PEDV-S1)/pVAX1-(IL-18) combination, an attenuated PEDV vaccine, or vector only control.Animals were examined for T lymphocyte proliferation, anti-PEDV antibodies, IFN-γ and IL-4 protein levels, and cytotoxic T cell function in mouse spleens and peripheral blood.In all cases, results showed that pVAX1-(PEDV-S) and the combination of pVAX1-(PEDV-S1) with pVAX1-(IL-18) induced the strongest responses but that pIL-18 had no adjuvant effects when given in combination with pVAX1-(PEDV-S1).