The eukaryotic expression plasmid of M2e-Hsp70 is a candidate M2e-based DNA vaccine. In order to evaluate the immunization potential of this construct, Specific Pathogen Free chickens were immunized either intramuscularly or orally. Mutant Salmonella typhimurium was used as carrier for oral delivery of the M2e-Hsp70 construct. M2e-specific humoral and cellular responses were tested by ELISA and Lymphocyte Proliferation Assay, respectively. Our results indicate that both humoral and cellular immune responses are conferred against M2e-Hsp70 plasmid in either of the intramuscular or oral routes of administration; however, these responses are significantly higher in intramuscular injection in contrast to oral administration. When it comes to mass vaccination of commercial chicken flocks oral administration is preferred due to the ease of application as well as its capability of eliciting mucosal, humoral and cellular immune responses; so measurements should be taken to improve the immunization potency of our orally delivered DNA vaccine.