Capsule Type of Determines Growth Phenotype

English version České info

The polysaccharide capsule of Streptococcus pneumoniae defines over ninety serotypes, which differ in their carriage prevalence and invasiveness for poorly understood reasons. Recently, an inverse correlation between carriage prevalence and oligosaccharide structure of a given capsule has been described. Our previous work suggested a link between serotype and growth in vitro. Here we investigate whether capsule production interferes with growth in vitro and whether this predicts carriage prevalence in vivo. Eighty-one capsule switch mutants were constructed representing nine different serotypes, five of low (4, 7F, 14, 15, 18C) and four of high carriage prevalence (6B, 9V, 19F, 23F). Growth (length of lag phase, maximum optical density) of wildtype strains, nontypeable mutants and capsule switch mutants was studied in nutrient-restricted Lacks medium (MLM) and in rich undefined brain heart infusion broth supplemented with 5% foetal calf serum (BHI+FCS). In MLM growth phenotype depended on, and was transferred with, capsule operon type. Colonization efficiency of mouse nasopharynx also depended on, and was transferred with, capsule operon type. Capsule production interfered with growth, which correlated inversely with serotype-specific carriage prevalence. Serotypes with better growth and higher carriage prevalence produced thicker capsules (by electron microscopy, FITC-dextran exclusion assays and HPLC) than serotypes with delayed growth and low carriage prevalence. However, expression of cpsA, the first capsule gene, (by quantitative RT-PCR) correlated inversely with capsule thickness. Energy spent for capsule production (incorporation of H3-glucose) relative to amount of capsule produced was higher for serotypes with low carriage prevalence. Experiments in BHI+FCS showed overall better bacterial growth and more capsule production than growth in MLM and differences between serotypes were no longer apparent. Production of polysaccharide capsule in S. pneumoniae interferes with growth in nutrient-limiting conditions probably by competition for energy against the central metabolism. Serotype-specific nasopharyngeal carriage prevalence in vivo is predicted by the growth phenotype.

Vyšlo v časopise: Capsule Type of Determines Growth Phenotype. PLoS Pathog 8(3): e32767. doi:10.1371/journal.ppat.1002574
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002574

Souhrn

Zdroje

1. JinPKongFXiaoMOftadehSZhouF 2009 First report of putative Streptococcus pneumoniae serotype 6D among nasopharyngeal isolates from Fijian children. J Infect Dis 200 1375 1380

2. BentleySAanensenDMavroidiASaundersDRabbinowitschE 2006 Genetic analysis of the capsular biosynthetic locus from all 90 pneumococcal serotypes. PLoS Genet 2 e31

3. MoxonEKrollJ 1990 The role of bacterial polysaccharide capsules as virulence factors. Curr Top Microbiol Immunol 150 65 85

4. WatsonDMusherD 1999 A brief history of the pneumococcus in biomedical research. Semin Respir Infect 14 198 208

5. BrueggemannAPetoTCrookDButlerJKristinssonK 2004 Temporal and geographic stability of the serogroup-specific invasive disease potential of Streptococcus pneumoniae in children. J Infect Dis 190 1203 1211

6. KronenbergAZucsPDrozSMuhlemannK 2006 Distribution and invasiveness of Streptococcus pneumoniae serotypes in Switzerland, a country with low antibiotic selection pressure, from 2001 to 2004. J Clin Micro 44 2032 2038

7. WeinbergerDTrzcinskiKLuYBogaertDBrandesA 2009 Pneumococcal capsular polysaccharide structure predicts serotype prevalence. PLoS Pathog 5 e1000476

8. BattigPHathawayLHoferSMuhlemannK 2006 Serotype-specific invasiveness and colonization prevalence in Streptococcus pneumoniae correlate with the lag phase during in vitro growth. Microbes Infect 8 2612 2617

9. BattigPMuhlemannK 2007 Capsule genes of Streptococcus pneumoniae influence growth in vitro. FEMS Immunol Med Microbiol 50 324 329

10. HathawayLBattigPMuhlemannK 2007 In vitro expression of the first capsule gene of Streptococcus pneumoniae, cpsA, is associated with serotype-specific colonization prevalence and invasiveness. Microbiology 153 2465 2471

11. IshizakaAWatanabeMYamashiaTOgawaYKohH 2001 New bronchoscopic microsample probe to measure the biochemical constituents in epithelial lining fluid of patients with acute respiratory distress syndrome. Crit Care Med 29 896 898

12. NelsonARocheAGouldJChimKRatnerA 2007 Capsule enhances pneumococcal colonization by limiting mucus-mediated clearance. Infect Immun 75 83 90

13. MoronaJPatonJMillerDMoronaR 2000 Tyrosine phosphorylation of CpsD negatively regulates capsular polysaccharide biosynthesis in Streptococcus pneumoniae. Mol Microbiol 35 1431 1442

14. BenderMCarteeRYotherJ 2003 Positive correlation between tyrosine phosphorylation of CpsD and capsular polysaccharide production in Streptococcus pneumoniae. J Bact 185 6057 6066

15. KadiogluABrewinHHärtelTBrittanJKleinM 2010 Pneumococcal protein PavA is important for nasopharyngeal carriage and development of sepsis. Mol Oral Microbiol 25 50 60

16. RichardsLFerreiraDMiyajiEAndrewPKadiogluA 2010 The immunising effect of pneumococcal nasopharyngeal colonisation; protection against future colonisation and fatal invasive disease. Immunobiology 215 251 263

17. KadiogluATaylorSIannelliFPozziGMitchellT 2002 Upper and lower respiratory tract infection by Streptococcus pneumoniae is affected by pneumolysin deficiency and differences in capsule type. Infect Immun 70 2886 2890

18. KadiogluAGinglesNGrattanKKerrAMitchellT 2000 Host cellular immune response to pneumococcal lung infection in mice. Infect Immun 68 492 501

19. MuhlemannKMatterHTauberMBodmerT 2003 Nationwide surveillance of nasopharyngeal Streptococcus pneumoniae isolates from children with respiratory infection, Switzerland, 1998–1999. J Infect Dis 187 589 596

20. SungCLiHClaverysJMorrisonD 2001 An rpsL cassette, Janus, for gene replacement through negative selection in Streptococcus pneumoniae. Appl Environ Microbiol 67 5190 5196

21. TrzcinskiKThompsonCLipsitchM 2003 Construction of otherwise isogenic serotype 6B, 7F, 14, and 19F capsular variants of Streptococcus pneumoniae strain TIGR4. Appl Environ Microbiol 69 7364 7370

22. MeierPUtzSAebiSMuhlemannK 2003 Low-level resistance to rifampin in Streptococcus pneumoniae. Antimicrob Agents Chemother 47 863 868

23. LacksS 1966 Integration efficiency and genetic recombination in pneumococcal transformation. Genetics 53 207 235

24. BrewsterJ 2003 A simple micro-growth assay for enumerating bacteria. J Microbiol Methods 53 77 86

25. XayarathBYotherJ 2007 Mutations blocking side chain assembly, polymerization, or transport of a Wzy-dependent Streptococcus pneumoniae capsule are lethal in the absence of suppressor mutations and can affect polymer transfer to the cell wall. J Bact 189 3369 3381

26. GoncalvesVMTakagiMLimaRBMassaldiHGiordanoRC 2003 Purification of capsular polysaccharide from Streptococcus pneumoniae serotype 23F by a procedure suitable for scale-up. Biotechnol Appl Biochem 37 283 287

27. AnumulaKR 1994 Quantitative determination of monosaccharides in glycoproteins by high-performance liquid chromatography with highly sensitive fluorescence detection. Anal Biochem 220 275 283

28. AnumulaKRDhumeST 1998 High resolution and high sensitivity methods for oligosaccharide mapping and characterization by normal phase high performance liquid chromatography following derivatization with highly fluorescent anthranilic acid. Glycobiology 8 685 694

29. SaddicGNEbertMBDhumeSTAnumulaKR 2002 Carbohydrate composition analysis of glycoproteins using highly sensitive fluorescence detection methods. Methods Mol Biol 194 23 36

30. StuderDGraberWAl-AmoudiAEggliP 2001 A new approach for cryofixation by high-pressure freezing. J Microsc 203 285 294

31. HunzikerEGraberW 1986 Differential extraction of proteoglycans from cartilage tissue matrix compartments in isotonic buffer salt solutions and commercial tissue-culture media. J Histochem Cytochem 34 1149 1153

32. HyamsCCamberleinECohenJBaxKBrownJ 2010 The Streptococcus pneumoniae capsule inhibits complement activity and neutrophil phagocytosis by multiple mechanism. Infect Immun 78 704 715

33. GatesMThorkildsonPKozelT 2004 Molecular architecture of the Cryptococcus neoformans capsule. Mol Microbiol 52 13 24