Bacterial surface lipoproteins mediate epithelial microinvasion byStreptococcus pneumoniae
Authors: Chan Jia Mun , Ramos-Sevillano Elisa , Betts Modupeh , Wilson Holly U. , Weight Caroline M. , Houhou-Ousalah Ambrine , Pollara Gabriele , Brown Jeremy S. , Heyderman Robert S. 
Published: October 2023

ABSTRACTStreptococcus pneumoniae, a common coloniser of the upper respiratory tract, invades nasopharyngeal epithelial cells without causing disease in healthy people. We hypothesised that surface expression of pneumococcal lipoproteins, recognised by the innate immune receptor TLR2, mediate epithelial microinvasion. Mutation oflgtin serotype 4 (TIGR4) and serotype 6B (BHN418) pneumococcal strains abolishes the ability of the mutants to activate TLR2 signalling. Loss oflgtalso led to concomitant decrease in interferon signalling triggered by the bacterium. However, only BHN418lgt::cmbut not TIGR4lgt::cmwas significantly attenuated in epithelial adherence and microinvasion compared to their respective wild-type strains. To test the hypothesis that differential lipoprotein repertoires in TIGR4 and BHN418 lead to the intraspecies variation in epithelial microinvasion, we employed a motif-based genome analysis and identified an additional 525 a.a. lipoprotein (pneumococcalaccessory lipoproteinA;palA) encoded by BHN418 that is absent in TIGR4. The gene encodingpalAsits within a putative genetic island present in ∼10% of global pneumococcal isolates. WhilepalAwas enriched in carriage and otitis media pneumococcal strains, neither mutation nor overexpression of the gene encoding this lipoprotein significantly changed microinvasion patterns. In conclusion, mutation oflgtattenuates epithelial inflammatory responses during pneumococcal-epithelial interactions, with intraspecies variation in the effect on microinvasion. Differential lipoprotein repertoires encoded by the different strains do not explain these differences in microinvasion. Rather, we postulate that post-translational modifications of lipoproteins may account for the differences in microinvasion.IMPORTANCEStreptococcus pneumoniae(pneumococcus) is an important mucosal pathogen, estimated to cause over 500,000 deaths annually. Nasopharyngeal colonisation is considered a necessary prerequisite for disease, yet many people are transiently and asymptomatically colonised by pneumococci without becoming unwell. It is therefore important to better understand how the colonisation process is controlled at the epithelial surface.Controlled human infection studies revealed the presence of pneumococci within the epithelium of healthy volunteers (microinvasion). In this study, we focused on the regulation of epithelial microinvasion by pneumococcal lipoproteins. We found that pneumococcal lipoproteins induce epithelial inflammation but that differing lipoprotein repertoires do not significantly impact the magnitude of microinvasion. Our results highlight the potential importance of the post-translational modification of lipoproteins in the mediation of epithelial invasion during pneumococcal colonisation. Targeting mucosal innate immunity and epithelial microinvasion alongside the induction of an adaptive immune response may be effective in preventing pneumococcal colonisation and disease.