Immunization with attenuated non-transformable pneumococcal pep27 and comD mutant provides serotype-independent protection against pneumococcal infection

Streptococcus pneumoniae is a well-known pathogenic bacterium with a high mortality rate. Currently, a 23-valent pneumococcal polysaccharide vaccine (PPV23) and protein-conjugate vaccines (PCVs) are available on the market. However, both of these vaccines have limitations; specifically, PPV23 produces weak antibody responses in children younger than 2 years and PCVs only partially protect against secondary infection. Previously, we showed serotype-nonspecific protection by Δpep27 vaccine, but the reversion of Δpep27 to the wild type serotype during immunization cannot be excluded. To ensure the safety of the Δpep27 vaccine, comD, an important protein that activates competence, was inactivated, and the transformability of the double mutant (Δpep27ΔcomD) was determined. The transformation ability of this double mutant was successfully abolished. Δpep27ΔcomD immunization significantly increased the survival time after heterologous challenge(s), and diminished colonization levels independent of serotype, including a non-typeable strain (NCC1). Moreover, the double mutant was found to be highly safe in both normal and immunocompromised mice. In conclusion, this pneumococcal Δpep27ΔcomD vaccine appears to be a highly feasible and safe vaccine to prevent various types of pneumococcal infections.