Streptococcus pneumoniae (S. pneumoniae, pneumococcus) is a major cause of morbidity and mortality worldwide. Achieving long-term immunity against S. pneumoniae through immunization is an important public health priority. Long-term protection after immunization is thought to rely both on protective serum antibody levels and immunological memory in the form of antigen-specific memory B cells (MBCs). Although the ability to achieve protective antibody levels shortly after pneumococcal vaccination has been well documented for the various infant immunization schedules currently in use worldwide, the examination of immunological memory in the form of antigen-specific MBCs has been much more limited. Such responses are critical for long-term protection against pneumococcal colonization and disease. This review summarizes the published literature on the MBC response to primary or booster immunization with either pneumococcal polysaccharide vaccine (PPV23) or pneumococcal conjugate vaccines (PCVs), aiming to elucidate the immunological mechanisms that determine the magnitude and longevity of vaccine protection against pneumococcus. There is evidence that PCVs induce the production of antigen-specific MBCs, whereas immunization with PPV23 does not result in the formation of MBCs. Increased understanding of the immunological factors that facilitate the induction, maintenance and recall of MBCs in response to pneumococcal vaccination could enable the use of MBC enumeration as novel correlates of protection against S. pneumoniae. Ongoing studies that examine MBC response to pneumococcal vaccination in high burden settings will be extremely important in our understanding of long-term protection induced by pneumococcal conjugate vaccines.

Antigen‐specific B cell identification by flow cytometry is crucial for investigating their immunophenotype, subset distribution, and kinetics post‐infection or immunization. Methods using biotinylated polysaccharide antigens have been described, but there is still room for improvement regarding sensitivity and applicability. The aim of this study was the development and validation of a multimer bead‐based method for detecting pneumococcal polysaccharide serotypes (PS)‐specific B cells following pneumococcal immunization. PS was chemically biotinylated and mounted on anti‐biotin beads, and labeled with phycoerythrin (PE)‐conjugated anti‐biotin antibody to form a PS‐multimer used for cell staining. Labeled beads were washed to remove excess fluorochrome and diminish non‐specific labeling and background noise. Optimal ratios of PS‐bead conjugate to PE and PS‐multimer to cells were determined with titration assays. Comparison between the PS‐multimer and a PS‐PE monomer revealed enhanced detection of PS‐specific cells and considerable signal amplification, attributed to the multimeric form of the detection probe and increased availability of antigen epitopes. To validate the specificity of the method, a competition assay using unbound PS was performed. Following pre‐incubation with increasing PS concentrations, detection of PS‐specific B cells with the PS‐multimer was inhibited in a stepwise manner. Pre‐incubation with excess PS completely blocked the fluorescent signal. This novel bead‐based flow cytometry approach is a sensitive method demonstrating high specificity. It generated enhanced signals, provided clear‐cut results, and was easily applicable, not requiring B cell pre‐enrichment. It could be modified to adapt other antigens of interest, especially polysaccharides and proteins that could be used to probe antigen‐specific B cell responses. The study of such responses may elucidate the underlying mechanisms involved in the establishment of long‐term protection, provide evidence‐based rationale for improving currently available vaccines and vaccination strategies, and pave the way for future vaccine development.

Pneumococcal vaccination schedules are traditionally assessed based on the antibody response. The Memory B Cell (MBC) response has been less studied, despite its role in the magnitude and longevity of protection. We compared the immune response to different vaccination schedules with the 13-valent Pneumococcal Conjugate Vaccine (PCV13) and investigated the relationship between MBCs and the antibody response. Total and pneumococcal serotype (PS)-specific MBCs, their subsets and PS-specific IgG antibodies induced by a 3 + 0 (group A), 2 + 1 (group B) or 3 + 1 (group C) schedule in healthy infants were studied before and 1 month after the last PCV13. The relatively immature IgM+IgD+ MBC subset was the predominant subset in all groups but was larger in group A compared to group B and group C, indicating that age might be a significant parameter of the composition of the MBC pool. PS-specific MBCs at baseline were higher in group A, but they increased significantly only in the groups receiving the booster schedules (groups B and C). PS-specific IgM-only MBCs at baseline positively corelated with the antibody response and the PS-specific swIg MBCs post-immunization. Our findings illustrate the importance of a booster dose for the enrichment of PS-specific immunological memory. IgM-only MBCs and swIg MBCs may serve as additional correlates of vaccine-induced protection.

Acute otitis media (AOM) is a common disease among children and can be complicated by otorrhea (AOMO). In 2010, the 13-valent Pneumococcal Conjugate Vaccine (PCV13) replaced the 7-valent vaccine (PCV7) in Greece. We aimed to describe the microbiological profile of bacterial ΑOMO among children younger than 16 years across the two PCV periods in a tertiary children’s hospital. Middle ear fluid cultures from 2418 children with AOMO were collected from 2007 to 2022. Otopathogens were isolated and tested for antimicrobial susceptibility. Data were compared between the PCV7- (2007–2011) and PCV13-period (2012–2019). The most common otopathogen over the 16-year period was S. pyogenes (35.4%), followed by H. influenzae (33.8%), S. pneumoniae (26.6%), and M. catarrhalis (4.1%). Pneumococcal resistance to cefotaxime and clindamycin significantly increased from 2% to 4.5% (p = 0.019) and 16.1% to 22.8% (p = 0.039), respectively. Resistance of H. influenzae to ampicillin increased from 6.3% to 13.9% (p < 0.001). A significant reduction in cotrimoxazole-resistant S. pneumoniae from 31% to 22.4% (p = 0.012), and in clindamycin-resistant and erythromycin-resistant S pyogenes, from 17.4% to 9.3% and 21.4% to 10.8%, respectively (p ≤ 0.001), was observed. During 2013–2022, 38 S. pneumoniae serotypes were identified among 250 isolates. Serotype 3 (27.2%) and 19A (13.2%) prevailed, followed by 19F (7%). The most common causes after the shift to PCV13 are S. pyogenes and H. influenzae. However, S. pneumoniae remains an important otopathogen with significant antimicrobial resistance. Serotype 3 was mostly detected, followed by 19A.