A recombinant protein comprising a listeriolysin O (LLO) protein, wherein the LLO protein comprises a mutation, or deletion of 2-11 amino acid residues within the cholesterol-binding domain (CBD) of the LLO protein, and wherein the recombinant protein exhibits a greater than 100-fold reduction in hemolytic activity relative to wild-type LLO. The recombinant protein may further comprise a heterologous peptide, such as an antigenic peptide.