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In Streptococcus pneumonia, phosphoenolpyruvate protein phosphotransferase (PtsA) is an intracellular protein of the monosaccharide phosphotransferase systems. Biochemical and immunostaining methods were applied to show that PtsA also localizes to the bacterial cell-wall. Thus, it was suspected that PtsA has functions other than its main cytoplasmic enzymatic role. Indeed, recombinant PtsA and anti-rPtsA antiserum were shown to inhibit adhesion of S. pneumoniae to cultured human lung adenocarcinoma A549 cells. Screening of a combinatorial peptide library expressed in a filamentous phage with rPtsA identified epitopes that were capable of inhibiting S. pneumoniae adhesion to A549 cells. The insert peptides in the phages were sequenced, and homologous sequences were found in human BMPER, multimerin1, protocadherin19, integrinβ4, epsin1 and collagen type VIIα1 proteins, all of which can be found in A549 cells except the latter. Six peptides, synthesized according to the homologous sequences in the human proteins, specifically bound rPtsA in the micromolar range and significantly inhibited pneumococcal adhesion in vitro to lung- and tracheal-derived cell lines. In addition, the tested peptides inhibited lung colonization after intranasal inoculation of mice with S. pneumoniae. Immunization with rPtsA protected the mice against a sublethal intranasal and a lethal intravenous pneumococcal challenge. In addition, mouse anti rPtsA antiserum reduced bacterial virulence in the intravenous inoculation mouse model. These findings showed that the surface-localized PtsA functions as an adhesin, PtsA binding peptides derived from its putative target molecules can be considered for future development of therapeutics, and rPtsA should be regarded as a candidate for vaccine development.

BACKGROUND: Kingella kingae, a normal component of the upper respiratory flora, is being increasingly recognized as an important invasive pathogen in young children. Genetic diversity of this species has not been studied. METHODS: We analyzed 103 strains from different countries and clinical origins by a new multilocus sequence-typing (MLST) schema. Putative virulence gene rtxA, encoding an RTX toxin, was also sequenced, and experimental virulence of representative strains was assessed in a juvenile-rat model. RESULTS: Thirty-six sequence-types (ST) and nine ST-complexes (STc) were detected. The main STc 6, 14 and 23 comprised 23, 17 and 20 strains respectively, and were internationally distributed. rtxA sequencing results were mostly congruent with MLST, and showed horizontal transfer events. Of interest, all members of the distantly related ST-6 (n = 22) and ST-5 (n = 4) harboured a 33 bp duplication or triplication in their rtxA sequence, suggesting that this genetic trait arose through selective advantage. The animal model revealed significant differences in virulence among strains of the species. CONCLUSION: MLST analysis reveals international spread of ST-complexes and will help to decipher acquisition and evolution of virulence traits and diversity of pathogenicity among K. kingae strains, for which an experimental animal model is now available.

BACKGROUND: Outbreaks and sporadic cases of pneumococcal illness occur among young adults in confined settings. Our aim was to characterize pneumococcal acquisition and carriage among healthy young adults in Israel during military training in confined settings. METHODS: During the years 2007-2008, an observational longitudinal study was conducted in three cohorts of healthy soldiers, during a 7-month basic training period. Epidemiological data, oropharyngeal and nasopharyngeal cultures were sampled on 5 occasions: before and 3, 6, 12 and 24 weeks after start of training. Samples were processed within 2-18 hours. Relatedness of isolates was investigated by capsular typing of all isolates and pulsed-field gel electrophoresis to determine acquisition and transmission. Carriage and acquisition patterns were analyzed and multivariable logistic regression analysis was performed to assess the impact of time on acquisition after mixing, controlling for other covariates. RESULTS: Pneumococci were recovered on 202 of 1872 visits among 742 individuals, including 40 different serotypes. Mean carriage prevalence increased in all visits following training initiation. Acquisition during training was high, as 36.9% of individuals acquired pneumococci at least once during training, and for almost one fourth of the whole population this occurred during the first 6 weeks. Significant clustering was noted. Sharing drinking glass/bottle was found to be a significant and common risk factor for pneumococcal acquisition. CONCLUSIONS: Pneumococcal acquisition is highly frequent when young adults live in close contact in confined settings, especially early after mixing.