Protection against systemic fatal pneumococcal infection by maternal intranasal immunization with pneumococcal surface protein A (PspA).

Streptococcus pneumoniae is a leading causative pathogen responsible for various types of bacterial infectious diseases in children. The aim of this study was to evaluate the protection conferred against fatal pneumococcal infections during infancy by maternal intranasal immunization with pneumococcal surface protein A (PspA). Four-week-old female BALB/c mice were immunized with PspA mixed with, or without, cholera toxin B (CTB) intranasally twice a week for 3 weeks. After the final immunization, they were mated with male mice to obtain offspring. Offspring at 10 days old were intraperitoneally inoculated with a pneumococcus strain, TIGR4, serotype 4. After the infections their survival periods were monitored. Anti-PspA-specific IgG antibody was induced in sera and breast milk at birth and maintained for 14 days during nursing periods in the PspA-immunized mother mice. At birth, offspring delivered from PspA-immunized mother mice had levels of anti-PspA-specific IgG antibody in sera same to those in their mothers on the day of birth. The survival times to death of offspring delivered from PspA-immunized mother mice after systemic fatal pneumococcal infections were significantly extended compared to those of controls. These findings suggest that maternal intranasal immunization with PspA could be an attractive procedure to employ against pneumococcal infections in early childhood.

Determination of pneumococcal serotypes/genotypes in nasopharyngeal secretions of otitis media children by multiplex PCR.

The appropriate clinical applications of pneumococcal polysaccharide vaccines against recent increases in antimicrobial resistant Streptococcus pneumoniae (S. pneumoniae) urgently require accurate analytical methodologies for determining and characterizing the serotypes. The results of current immunological determinations of serotypes with anti-capsular polysaccharide-specific sera are difficult to interpret in terms of quellung changes of the pneumococci. In this study, we applied the multiplex PCR technique for the rapid identification of pneumococci and simultaneous rapid determinations of their serotypes and genotypes that directly correlated with antimicrobial susceptibilities from nasopharyngeal secretions (NPS). Serogroups 6, 19F and 23F were the predominant capsular types of S. pnuemoniae in the NPS samples. Strains of serotypes 19F and 23F frequently had mutations in pbp1a, pbp2x and pbp2b and expressed ermB and mefA; they also were mostly resistant to both penicillin G (PCG) and clarithromycin (CAM). Two NPS samples contained the strain of serotype 19F together with the strain of serotype 23F, although only the strain of serotype 19F was identified by a conventional bacterial culture. Pneumococci were identified in six NPS samples and their serotypes determined by the multiplex PCR, while a conventional bacterial culture failed to identify the pathogens. Our findings suggest that PCR-based serotyping and genotyping can provide an accurate and rapid distribution of pneumococcal serotypes and antimicrobial resistance. The relatively minor populations in the nasopharynx may be determined using molecular techniques.