Pathogenesis of Streptococcus pneumoniae serotype 3 during natural colonization and infections among children and its IgG correlate of protection in a mouse model.

Current licensed pneumococcal conjugate vaccines (PCVs) are effective against pneumococcal diseases caused by the serotypes contained in the PCvs However; several studies evaluating pneumococcal colonization and acute otitis-media (AOM) prevention in young children vaccinated with PCV13, observed less effectiveness against serotype-3. One possible reason for less effectiveness may be release of the capsular polysaccharide (CPS) of serotype-3 (CPS-3) as an immune evasion mechanism. Here we evaluated free CPS-3 levels released from 6 clinical isolates from young children compared to WU2 strain and to serotype-19A CPS (CPS-19A) released in vitro when interacting with nasopharyngeal, middle-ear and lung cell-lines. Clinical serotype-3 strains showed greater release of CPS than WU2 with the interaction to 2 cell-lines and all 6 clinical serotype-19A strains. We next evaluated CPS-3 vs CPS-19A levels in middle-ear fluid (MEF) and the nasopharynx (NP) of young children and found higher levels of CPS-3 compared to CPS-19A in MEF during AOM but not in NP secretions during colonization. With anti-CPS-3 IgG in MEF and NP secretions at time of health and onset of AOM, a significant negative correlation (r = -0.75, p < 0.05) between unbound anti-CPS-3 IgG levels and free- anti-CPS-3 in MEF were found, and a significant lower detection of unbound anti-CPS-3 IgG in NP at the time of health with serotype-3 SPN (p < 0.05) compared to irrelevant SPN serotypes were found. In a mouse model of AOM and pneumonia, we sought a correlate of protection against serotype-3 infection using human serum-derived anti-CPS-3 IgG. We conclude that serotype-3 clinical isolates from children release more capsule than WU2 strains or 19A strains during in vitro testing; release more capsule in the MEF of children during AOM than serotype 19A; unbound anti-CPS-3 IgG levels negatively correlate with free-anti-CPS-3; and a level of 2.8 µg/ml anti-CPS-3 antibody protects mice from AOM and pneumonia but not colonization.