ISSN 2045-2322 (online). AliA protein of strains D39 and R6 are identical but AmiA protein differs by one amino acid in the predicted binding site, and AliA peptide suppressed growth of D39 but not R6. However, a pathotype of K. pneumoniae, termed hypervirulent K. pneumoniae (hv Kp ), has emerged and is spreading throughout the community, causing severe, often fatal, disease in healthy individuals. Note that the only sequence difference between D39 and S. pseudopneumoniae (inhibited by AmiA peptide ligand) and R6 and S. mitis (not inhibited by AmiA peptide ligand) is at position 54 (indicated by blue arrow), predicted to be within the binding site. Results represent 3 independent experiments, error bars indicate SEM. However, the analysis revealed which amino acids are likely to be at the peptide-protein interaction interface. Bacteria pellets were re-suspended in 50l lysis buffer (8M Urea/100mM TrisHCl pH8, with Roche complete protease inhibitor cocktail) and sonicated with two short bursts of 5s at 10s intervals on ice. Tables 1 and 2 illustrate the demographics, systemic illnesses, clinical features, management details, and visual outcomes of patients with S. pneumoniae endophthalmitis. The Institutional Review Board of Chang Gung Memorial Hospital in Taoyuan, Taiwan approved this retrospective study protocol and waived the need for written informed consent from the study patients. Kuan-Jen Chen. Scientific Reports (Sci Rep) Clin. Robust label-free, quantitative profiling of circulating plasma microparticle (MP) associated proteins. Ophthalmol. L.J.H. Sign up for the Nature Briefing newsletter what matters in science, free to your inbox daily. https://doi.org/10.1038/s41586-021-03819-2 (2021). Statistical analysis was performed using SPSS for Windows version 23 (SPSS Science, Chicago, IL, USA). Schaffner, T. O. et al. Cell. This amino acid difference at position 54 between strains D39 and R6 confirmed the finding of Lanie et al.24. You will be subject to the destination website's privacy policy when you follow the link. Natl. Infect. S. pneumoniaehas various virulence factors, including polysaccharide capsules, pneumolysin, neuraminidases, and zinc metalloproteinases. Rev. Although S. pneumoniae isolates were susceptible to vancomycin, S. pneumoniae endophthalmitis had a very poor visual prognosis. 11, 183. https://doi.org/10.3389/fnins.2017.00183 (2017). Pneumonia is one of the most serious diseases for children, with which lung microbiota are proved to be associated. The need for a higher number of intravitreal injections was significantly associated with a poor final VA7. The peptide ligands of AmiA, AliA and AliB proteins determine pneumococcal phenotypes, such as pneumococcal growth, biofilm production, capsule size, chain length and transformation rate13. J. Ophthalmic Inflamm. It is especially problematic in hospitals, where it causes a range of acute infections. They are usually found in pairs and are nonsporingand nonmotile. Klebsiella spp. The protein concentration was measured by Qubit Protein Assay (Invitrogen). Amino acids highlighted in grey are part of the N-terminal signal peptide. S. pneumoniae resides asymptomatically in healthy carriers, and they typically colonize the respiratory tract, sinuses, andnasal cavity. The staining was performed as described in the manufacturer's protocol. 5, 503. https://doi.org/10.1038/s42003-022-03445-2 (2022). We did this by predicting the peptide-protein interaction interface with PepNN19, shown in Fig. 33, 671674 (2018). https://doi.org/10.1038/s41592-022-01488-1 (2022). (K group, n=76) and S. pneumoniae (S group, n=446) were included. 122, 117 (1996). The most common etiology in the current series was postoperative endophthalmitis; this finding is similar to those in studies conducted in the United States, Europe, Australia, and Thailand5,6,7,8,9,10. include pneumonia, lung abscess, and empyema. Alloing, G., de Philip, P. & Claverys, J.-P. Three highly homologous membrane-bound lipoproteins participate in oligopeptide transport by the ami system of the gram-positive Streptococcus pneumoniae. You can review and change the way we collect information below. Thank you for taking the time to confirm your preferences. All patients with postcataract endophthalmitis developed acute-onset endophthalmitis (median, 3days; range, 1 to 8days). The treatment strategies were determined by the individual treating physicians and did not follow a standardized protocol. J. Proteome Res. 34, 184193. Am. The cultures were incubated in a water bath at 37C and dilutions streaked out on CSBA plates at the timepoint of inoculation, after 3 and 4.5h. The CSBA plates were incubated at 37C and 5% CO2 and colonies counted the next day. The main clinical features were postcataract endophthalmitis (n=13, 34%) and endophthalmitis associated with corneal ulcer (n=12, 32%), trauma (n=6, 16%), endogenous etiology (n=4, 11%), trabeculectomy (n=2, 5%), and pterygium excision-related scleral ulcer (n=1, 3%). Visualization in the posterior segment view was poor in all eyes due to severe anterior chamber inflammation and severe vitritis. This information was used in the following sections to determine whether any amino acid difference between strains or species might affect binding sites. A study in Semarang, Indonesia, found nasopharyngeal colonization of K. pneumoniae in 7% of healthy children (660 months old) and 15% of adults (4570 years old)8. Positive reactions included optochin, bile solubility, inulin and alfa hemolysis, and negative reactions included catalase and oxidase. Monocultures were prepared by combining 500l of CDM with 500l of bacterial suspension, co-cultures by combining 500l of the K. pneumoniae and 500l of the S. pneumoniae bacterial suspensions. The samples were acidified to a concentration of 0.1% in formic acid. If you need to go back and make any changes, you can always do so by going to our Privacy Policy page. This study investigated the clinical details, antibiotic susceptibility, and visual outcomes of patients with culture-proven endophthalmitis caused by S. pneumoniae infection at a tertiary referral center in Taiwan. 187, 83408349. On the binding mechanism of the peptide receptor of the oligopeptide transport system of Lactococcus lactis. Hilty, M. et al. Visual acuity (VA) test results were as follows: hand motions (HM; n=14/38, 37%), light perception (LP; n=12/38, 32%), no light perception (NLP; n=7/38, 18%), counting fingers (4/38, 11%), and undetermined VA in one 2-year-old patient. In other studies, the most common causes of S. pneumoniae endophthalmitis were surgery and trauma5,9,10,11,12. Genomic DNA was extracted from S. pneumoniae strains D39, R6 and ATCC 17619 with the QIAamp DNA Mini Kit (Qiagen) according to the manufacturer's protocol. Front. 14, 210210. They also have a high tendency to. Conclusion: Human spermatozoa act as . S3). Cookies used to make website functionality more relevant to you. J. Ophthalmic Inflamm. Clinical strains of S. pneumoniae of various genetic backgrounds were used (serotypes in brackets): 111.46 (19F), 202.58 (1), 208.41 (7F), 307.14 (18C). These cookies perform functions like remembering presentation options or choices and, in some cases, delivery of web content that based on self-identified area of interests. Chen, K. J. et al. This made statistical analysis difficult to perform. PubMedGoogle Scholar. Sensing peptides derived from other bacteria and having steady but persistent growth might be an advantage in colonization in a hostile environment or when nutrient availability is limited13. Streptococcus pneumoniae (pneumococcus) is aGram-positive, spherical bacteria. In conclusion, the most common cause in S. pneumoniae endophthalmitis infection was postcataract status. Lux, J., Holivololona, L., San Millan Gutierrez, R. et al. Article The peptide-protein interaction interface was visualized with PyMol (The PyMol Molecular Graphics System, Version 2.5.2 Schrdinger, LLC). The Centers for Disease Control and Prevention (CDC) cannot attest to the accuracy of a non-federal website. https://doi.org/10.1093/femsre/fuy043 (2018). Amino acids highlighted in red were predicted to be at the AliA peptide ligand-AliA protein interaction interface by PepNN, interaction propensity/probability of0.5. Methods 14, 513520. Protein abundance ratios between strains ATCC 17619 and D39 were calculated based on the sum of all peptide MS1 signals from DDA and averaged MS2 fragment signals from DIA data. Image Copyright: ktsdesign / Shutterstock S. pneumoniae is. The slides were viewed using a Zeiss Axio Imager M1 fluorescence microscope with a 40objective. Centers for Disease Control and Prevention. Presenting visual acuity ranged from counting fingers to no light perception. The amino acids of AmiA and AliA that are predicted to be at the peptide-protein binding interface are located in what we hypothesize are the binding pockets. AlphaFold models of oligopeptide-binding proteins AmiA and AliA from 110.58 were used together with the sequence of AmiA and AliA peptide ligand respectively to predict the peptide-binding interface with PepNN. Podschun, R., Pietsch, S., Hller, C. & Ullmann, U. C. collected and analyzed the data; H.-C. C.,C.-C. L., M.-H. S., Y.-P. C., N.-K. W., Y.-S. H., A.-N. C., W.-C. W., L. Y., C.-C. S., L. L., Y.-H. C. and H.-D. C. all provided patients data and assisted in data collection. Am. Mol. ADS In the endogenous etiology group, endogenous causes included infective endocarditis and pneumonia, and two patients had unidentified primary sources of infection. Data collected and reviewed included demographic information, medical history, presenting signs and symptoms, duration of symptoms before diagnosis of endophthalmitis, intervals between event and diagnosis of endophthalmitis, culture sites, antibiotic sensitivities and resistance patterns, management administered, and final VA. Intraocular fluids were collected and plated on blood agar (5% sheep), chocolate agar, fungus culture media, and thioglycolate broth. PubMedGoogle Scholar. Yospaiboon, Y. et al. PubMed Central Doctors consider some of these infections invasive. Invasive disease means that germs invade parts of the body that are normally free from germs. Chen, K. J. et al. We thank Carlo Casanova (Institute for Infectious Diseases, University of Bern, Switzerland) for kindly providing reference strains of H. influenzae, S. aureus, M. catarrhalis and K. pneumoniae. Get the most important science stories of the day, free in your inbox. Jindal, A. et al. https://doi.org/10.1186/s12866-014-0210-x (2014). S. pneumoniaeis still a main cause of ocular infectious diseases, such as conjunctivitis, keratitis, and endophthalmitis1,2. In three patients with polymicrobial endophthalmitis, Proteus mirabilis, Klebsiella oxytoca, and Serratia marcescens were identified. In addition, we previously proposed that pneumococcus can detect bacteria within the same microbiota via short peptide fragments derived from other bacterial species that bind to the Ami-AliA/AliB permease11,12,13,14. wrote the first draft of the manuscript. Details of all genomic differences between D39 and R6 strains are shown in Supplementary Table S3. Experts refer to them as Gram-negative, encapsulated, and nonmobile bacteria. Streptococcus pneumoniae endophthalmitis: clinical settings, antibiotic susceptibility, and visual outcomes. In the present study, only three eyes (3 out of 38, 8%) achieved vision of 20/400 or better. Biol. Nat. CDC is not responsible for Section 508 compliance (accessibility) on other federal or private website. Environ. 2). Ophthalmol. All research was performed in accordance with the relevant guidelines and regulations. Patients with endophthalmitis caused by S. pneumoniae or P. aeruginosa infection have a higher risk of requiring evisceration or enucleation13,15. Mirdita, M. et al. Streptococcus pneumoniae detects and responds to foreign bacterial peptide fragments in its environment. Concentrations of peptide ranged from 0 to 0.5 mg/ml. Correspondence to Get the most important science stories of the day, free in your inbox. 5a), are highlighted in red in the multiple sequence alignment in Fig. Chen et al.16 demonstrated that prevention of evisceration or enucleation in patients with endogenous bacterial panophthalmitis, no light perception, and scleral abscess can be achieved through multiple intravitreal and periocular injections of antibiotics and dexamethasone. Here, we aimed to determine whether the AmiA, AliA and AliB peptide ligands are indeed secreted by K. pneumoniae. Bacterial culture isolates were identified through conventional microbiological methods (20002013) and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOFMS; 20142019). The sequence EMQPIVDRQ was included in 7 out of these 8 peptides and therefore seems to be a consensus sequence. Kerr, A. R. et al. Hathaway, L. J. et al. Klebsiella pneumoniae is a member of the Klebsiella genus of Enterobacteriaceae and belongs to the normal flora of the human mouth and intestine. Sci. This causes significant morbidity and mortality annually, especially in young children, the elderly and immunocompromised2. Ophthalmol. Front. https://doi.org/10.1093/emboj/19.14.3649 (2000). Proteus mirabilis, Klebsiella . Benton, A. H. & Marquart, M. E. The role of pneumococcal virulence factors in ocular infectious diseases. Clinical presentations, microbiology and management outcomes of culture-proven endogenous endophthalmitis in India. https://doi.org/10.1128/JB.182.6.1600-1608.2000 (2000). Pneumococcal reference strains ATCC 700903, ATCC 17619, D39 and R6 were also used28,29 and a D39 knock out mutant of AmiA (D39AmiA)13,16, as were reference strains of the other bacterial species: H. influenzae ATCC 49247, S. aureus ATCC 29213, M. catarrhalis ATCC 8176 and Klebsiella pneumoniae ATCC BAA-1706. 1), indicating that K. pneumoniae had reduced the growth of S. pneumoniae. Methods 53, 7786. We speculated that there must be mutations in other parts of the R6 genome that affect the signaling of the AliA peptide ligand and the growth-inhibitory effect. Primary evisceration was performed on six eyes, and secondary evisceration or enucleation was performed on four eyes. The bacteria may be isolated from the nasopharynx of 5-90% of healthy persons, depending on the population and setting: 5-10% of adults without children are carriers 20-60% of school-aged children may be carriers This work was funded by grant number 192067 from the Swiss National Science Foundation to L.J.H. . Han, D. P. et al. For AliA protein we could not identify a single amino acid change causing loss of binding although we did note amino acid changes in the AliA protein of S. pneumoniae ATCC 17619, S. pseudopneumoniae and S. mitis compared to D39. PubMed Central A study on the peptide receptor of the Opp oligopeptide transport system (Ami-AliA/AliB homologue) of Lactococcus lactis suggests that small peptides are completely enclosed by the binding protein, whereas longer peptides might partially protrude from the binding pocket21. By mass spectrometry, we identified such peptides in the Klebsiella pneumoniae secretome. Cite this article. Because of the limitations inherent in a retrospective study, the antibiotic susceptibility testing of S. pneumoniae isolates was performed in some selected antibiotics, including vancomycin, penicillin, ceftriaxone, levofloxacin, and moxifloxacin. 4). Streptococcus pneumoniae endophthalmitis is clinically more severe, more difficult to treat, and carry a higher risk of vision loss, evisceration, or enucleation. Enucleation or evisceration was performed when the globe was not salvageable due to corneal or scleral necrotic laceration. In other studies of streptococcal endophthalmitis, S. pneumoniae led to the worst visual outcomes out of the culture-proved organisms examined3,5,6,7,8. Growth was measured in peptide-free chemically defined medium (CDM) by measuring optical density (OD) over time. Our study demonstrated that the MIC90 of vancomycin for S. pneumoniae was 0.5g/mL. INTRODUCTION. Google Scholar. The *.pdb files obtained together with the AmiA or AliA peptide ligand sequence were used to predict the peptide-protein interaction interface with PepNN-Struct. The data for three biological replicates was analyzed. Some have capsules surrounding their cells,. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate. The percent homologies on nucleotide and amino acid level compared to strain D39 are shown in Table 3. Streptococcus pneumoniae endophthalmitis is clinically more severe, more difficult to treat, and carry a higher risk of vision loss, evisceration, or enucleation. Vaccines 8, 132. https://doi.org/10.3390/vaccines8010132 (2020). Historically, K. pneumoniae has caused serious infection primarily in immunocompromised individuals, but the recent emergence and spread of hypervirulent strains have broadened the number of people susceptible to infections to include those who are healthy . https://doi.org/10.1128/JB.01148-06 (2007). 8, 1012. https://doi.org/10.1371/journal.pone.0169603 (2017). 9, 320. https://doi.org/10.3389/fcimb.2019.00320 (2019). The structural basis of peptide-protein binding strategies. We then used the protein structure files and the AmiA or AliA peptide ligand sequences to predict the localization of putative peptide binding sites. When the isolates were identified through MALDI-TOF MS (Bruker Daltonics, Germany), the microbial film was overlaid with 1 L of 70% formic acid. Therefore, we tested whether AmiA and AliA peptide ligands were able to inhibit growth of genetically diverse clinical pneumococcal isolates, including antibiotic-resistant strains. 241, 4458. performed the experimental research, J.L. In terms of sex, 25 of the patients (66%) were male and 13 patients (34%) were female. Dave, T. V. et al. BLAST analysis indicated that AmiA and AliA proteins are highly conserved between pneumococcal strains. Pars plana vitrectomy with intravitreal antibiotics was performed in 17 eyes (39%) in primary or secondary treatments. Archiv fr pathologische Anatomie und Physiologie und fr klinische Medicin 87, 319324 (2005). Jumper, J. et al. J. Bacteriol. STAPH/STREP COMBINATION- klebsiella pneumoniae, enterococcus faecalis, pneumococcinum, streptococcus agalactiae, strepto. Klebsiella pneumoniae bacteremia was defined as the isolation of K pneumoniae in 1 or more blood cultures in association with the clinical features of bacteremia and/or sepsis. Antibiotic susceptibility testing and minimal inhibitory concentration (MIC) breakpoints of antibiotics are summarized in Tables 3 and 4. We therefore propose their potential value as future therapeutic agents against pneumococcal diseases. https://doi.org/10.1073/pnas.0702377104 (2007). Kurniawan, E. D., Rocke, J. R., Sandhu, S. S. & Allen, P. J. Predictors of visual outcome and the role of early vitrectomy in streptococcal endophthalmitis. Biomol. Performance standards for antimicrobial susceptibility testing; eighteenth informational supplement. One patient with scleral ulcerrelated endophthalmitis underwent a complicated pterygium excision with subsequent scleral grafts. To obtain https://doi.org/10.1016/j.str.2009.11.012 (2010). PLoS ONE 12, e0169603. M.Hi. 5l of stained bacteria suspension were pipetted onto a microscope slide and the coverslip added firmly. Data acquisition was in data-dependent mode on the top 15 peaks with mass-to-charge ratio (m/z) between 360 and 1400, charge state 27, and no exclusion time set. Presenting symptoms of pain and decreased VA were identified in 11 of 13 eyes (96%) and in 13 of 13 eyes (100%) and had mean durations of 1.9 and 2.1days, respectively. 9, 9. https://doi.org/10.1186/s12348-019-0174-y (2019). Swiss clinical S. pneumoniae strains for this study originated from a strain collection obtained from a nationwide surveillance for nasopharyngeal S. pneumoniae isolates from children with respiratory infection27. AmiA and AliA peptide ligands are secreted by Klebsiella pneumoniae and inhibit growth of Streptococcus pneumoniae Janine Lux, Lalaina Holivololona, Raquel San Millan Gutierrez, Markus. Ten eyes (26%) underwent evisceration or enucleation. and JavaScript. Correspondence to One patient with traumatic endophthalmitis due to IOFB underwent removal of an anterior chamber IOFB and anterior chamber irrigation, and they were given antibiotics intravitreally. J. Mol. After the sample had dried again, microorganism identification and data analysis were performed, for which the Bruker LT Microflex MALDI-TOF MS and Bruker Biotyper 3.0 system software programs were used. Carriage of pneumococci Pneumococci are common inhabitants of the respiratory tract. In the meantime, to ensure continued support, we are displaying the site without styles Google Scholar. Cultures from 35 eyes (92%) yielded a single microorganism. Search parameters were as follows: precursor and fragment mass tolerances of 10ppm and 0.4Da, isotope error on first and second isotope, strict trypsin cleavage rule with a maximum of two missed cleavages per peptide, oxidation of Met and protein N-terminal acetylation as variable and fixed carbamidomethylation on Cys modifications. Whole Genome Shotgun project has been deposited at DDBJ/ENA/GenBank under accessions JAMLJP000000000 (S. pneumoniae ATCC 17619), JAMLJQ000000000 (S. pneumoniae R6), and JAMLJR000000000 (S. pneumoniae D39). Nasher, F., Kwun, M. J., Croucher, N. J., Heller, M. & Hathaway, L. J. Peptide occurring in enterobacteriaceae triggers Streptococcus pneumoniae cell death. At exponential growth phase OD600nm of 0.5, the analysis timepoint for peptide extraction, K. pneumoniae cells were stained using the LIVE/DEAD BacLight assay kit (Thermo Fisher Scientific). This study is to investigate the clinical settings, antibiotic susceptibility, and visual outcomes of S. pneumoniae endophthalmitis at a tertiary referral center in Taiwan. Klebsiella pneumoniae, a Gram-negative bacterium, was first described following isolation from the lungs of a pneumonia patient in 18824.
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