Although there has been great progress in treatment and prevention of infectious diseases in the past years, effective vaccines against some of the most serious infections, are urgently needed. The vesicular stomatitis virus (VSV) has proven to be a potent viral vector vaccine candidate.However, translation of these promising studies into clinical application has so far been limited by VSVs neurotropism. VSV pseudotyped with the glycoprotein GP of the lymphocytic choriomeningitis virus (VSV-GP) has completely lost neurotropism and is perfectly safe in mouse models, while maintaining replication fitness.Here, the potential of VSV-GP as a vaccine vector was analyzed with ovalbumin (OVA) as a model antigen. VSV-GP-OVA was directly compared to the VSV encoding OVA (VSV-OVA). In mice, VSV-OVA and VSV-GP-OVA induced equal levels of OVA-specific humoral and cellular immune responses upon a single immunization. However, boosting with the same vector was only possible for VSV-GP-OVA, as this vector did not induce vector specific humoral immunity, even after multiple applications. In contrast, neutralizing antibodies limited the immunogenicity of the VSV-OVA boost. In addition, OVA specific CTL responses induced by VSV-GP-OVA were at least as potent as those observed for a gold-standard adenoviral vector. Prophylactic vaccination with VSV-GP-OVA was highly efficient and completely protected mice in a Listeria monocytogenes challenge model.Taken together, VSV-GP can serve as a platform technology for the development of novel vaccines against a broad spectrum of diseases.