Various strategies for the production of safe vaccines have been used. This study compared three different inactivation procedures, i.e. treatment with formaldehyde, beta-propiolactone or UV-light using vesicular stomatitis virus (VSV) as a model antigen. All three inactivation procedures drastically impaired induction of neutralizing IgG responses, confirming previous observations [Bachmann et al. (1993) J Virol 67:3917-3922]. This reduction could be overcome using higher doses for all three preparations. Both formaldehyde and beta-propiolactone completely abrogated the induction of VSV-specific cytotoxic T cells (CTLs), whereas UV-inactivated virus was able to induce significant and long-lasting CTL responses. These results may be of practical relevance since induction of neutralizing antibodies alone is often not sufficient for protection and sometimes may even enhance immunopathological responses of vaccinees.