pneumoniae has been observed to form biofilms both in vitro and in vivo [9, Akt inhibitor 12–14, 24, 30, 33, 34]; although during invasive disease, pneumococci in the bloodstream and sputum seem to be exclusively diplococci. While a large body of work has been published on the characteristics of pneumococcal biofilm formation in vitro as well as the genes involved in this process, little is known about the host immune response to pneumococcal

biofilms and how this differs with respect to planktonic bacteria. This is a significant lapse as pneumococcal biofilms are now recognized to be present in the nasopharynx of colonized selleck inhibitor humans. In the present study, we identified the differential protein profile of S. pneumoniae serotype 4, strain TIGR4 in a mature 3-day old biofilm versus during planktonic exponential growth. As expected, we observed considerable differences in the protein profiles of planktonic and biofilm TIGR4 with the vast majority of detected proteins being produced in diminished quantities. Notably, our proteomic findings are in disagreement with those of Allegrucci et al. which described a dramatic increase in the number of detectable proteins in 9 day-old biofilms including phosphoglyceromutase, phosphoglycerate kinase, 30S ribosomal protein S1, translation elongation factor Tu, 50S ribosomal protein

L1, enolase, DnaK protein, and pyruvate oxidase, among many other proteins [24]. This discrepancy may be due to the different strains used, the different age Epigenetics inhibitor of the biofilms examined, alternatively, due to our strict criteria

for protein identification combined with the fact that that a large portion of mature biofilm is Tangeritin composed of dead and presumably degraded bacterial components. Importantly, our findings are in agreement with the generally accepted notion that the synthetic and metabolic activity of bacteria are reduced during biofilm growth [15, 16], as well as with previous studies examining the transcriptional changes incurred during pneumococcal biofilm growth which showed down-regulation of the genes encoding many of these proteins [17, 25, 30, 35]. Due to the altered protein profiles, unsurprisingly, but also previously undocumented, convalescent sera only robustly recognized planktonic cell lysates. Likewise, sera from biofilm-immunized mice weakly recognized cell lysates from planktonic pneumococci. Together, these results support the notion that invasive pneumococcal disease is predominantly caused by the planktonic phenotype. They also suggest that the antibody response and potentially the T-cell response generated against S. pneumoniae during nasopharyngeal colonization would be of limited utility against planktonic bacteria during invasive disease. This latter notion is supported by our finding that immunization with ethanol-killed TIGR4 biofilm pneumococci failed to protect against invasive disease caused by a serotype 3 isolate.