Classical swine fever (CSF), which is caused by classical swine fever virus (CSFV), causes significant losses in pig industry in many countries in Asia and Europe. The E2 glycoprotein of CSFV is the main target for neutralizing antibodies. In this study, a recombinant replication-defective human adenovirus expressing the CSFV E2 gene (rAdV-E2) was generated and evaluated for the immunogenicity in rabbits. The results showed that the rabbits immunized with rAdV-E2 developed high-level CSFV-specific antibodies. The rAdV-E2-immunized rabbits were all free of the regular fever and the viral replication in the spleen upon challenge with C-strain, which were seen in the rabbits immunized with the parent adenovirus of rAdV-E2. This indicates that the recombinant adenovirus can be an attractive candidate vaccine against CSF.