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. 1993 May;61(5):1743–1749. doi: 10.1128/iai.61.5.1743-1749.1993

Effect of growth conditions on expression and antigenicity of Staphylococcus epidermidis RP62A cell envelope proteins.

K P McDermid 1, D W Morck 1, M E Olson 1, M K Dasgupta 1, J W Costerton 1
PMCID: PMC280760  PMID: 8478063

Abstract

Staphylococcus epidermidis RP62A (ATCC 35984) was grown in tryptic soy broth (TSB), iron-depleted TSB (TSB-Fe), iron-reconstituted TSB-Fe (TSB+Fe), a chemically defined medium, and fetal calf serum (FCS) and on silastic disks in chambers that were sutured to the pig peritoneal wall. Bacterial cell wall proteins were extracted by digestion with recombinant lysostaphin, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and detected by silver staining. Cell wall proteins from TSB-, chemically defined medium-, or FCS-grown cells had a complex profile of greater than 25 protein bands spanning the full molecular mass range. By contrast, a digest obtained from in vivo-grown cells had only five major proteins of 40 kDa or greater. Proteins of 130 and 106 kDa were present in the cell envelopes of TSB-Fe- and in vivo-grown cells but not in those grown in TSB or TSB+Fe. A 43-kDa protein expressed by in vitro-grown cells and 52- and 96-kDa proteins expressed by in vivo-grown cells reacted with antisera from pigs with the chamber implants and from catheterized, paracatheter-inoculated pigs but not with hyperimmune sera from pigs immunized with TSB-grown cells. The data indicate that S. epidermidis, growing under in vivo conditions, expresses antigens distinct from those that are grown in vitro.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aitchison E. J., Lambert P. A., Farrell I. D. Antigenic composition of an endocarditis-associated isolate of Streptococcus faecalis and identification of its glycoprotein antigens by ligand blotting with lectins. J Med Microbiol. 1986 Mar;21(2):161–167. doi: 10.1099/00222615-21-2-161. [DOI] [PubMed] [Google Scholar]
  2. Bayer A. S., Speert D. P., Park S., Tu J., Witt M., Nast C. C., Norman D. C. Functional role of mucoid exopolysaccharide (alginate) in antibiotic-induced and polymorphonuclear leukocyte-mediated killing of Pseudomonas aeruginosa. Infect Immun. 1991 Jan;59(1):302–308. doi: 10.1128/iai.59.1.302-308.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Cheung A. L., Fischetti V. A. Variation in the expression of cell wall proteins of Staphylococcus aureus grown on solid and liquid media. Infect Immun. 1988 May;56(5):1061–1065. doi: 10.1128/iai.56.5.1061-1065.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Christensen G. D., Baddour L. M., Madison B. M., Parisi J. T., Abraham S. N., Hasty D. L., Lowrance J. H., Josephs J. A., Simpson W. A. Colonial morphology of staphylococci on Memphis agar: phase variation of slime production, resistance to beta-lactam antibiotics, and virulence. J Infect Dis. 1990 Jun;161(6):1153–1169. doi: 10.1093/infdis/161.6.1153. [DOI] [PubMed] [Google Scholar]
  6. Christensen G. D., Baddour L. M., Simpson W. A. Phenotypic variation of Staphylococcus epidermidis slime production in vitro and in vivo. Infect Immun. 1987 Dec;55(12):2870–2877. doi: 10.1128/iai.55.12.2870-2877.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Costerton J. W., Cheng K. J., Geesey G. G., Ladd T. I., Nickel J. C., Dasgupta M., Marrie T. J. Bacterial biofilms in nature and disease. Annu Rev Microbiol. 1987;41:435–464. doi: 10.1146/annurev.mi.41.100187.002251. [DOI] [PubMed] [Google Scholar]
  8. Finlay B. B., Falkow S. Common themes in microbial pathogenicity. Microbiol Rev. 1989 Jun;53(2):210–230. doi: 10.1128/mr.53.2.210-230.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gorby G. L., Robinson E. N., Jr, Barley L. R., Clemens C. M., McGee Z. A. Microbial invasion: a covert activity? Can J Microbiol. 1988 Apr;34(4):507–512. doi: 10.1139/m88-087. [DOI] [PubMed] [Google Scholar]
  10. Hogt A. H., Dankert J., Hulstaert C. E., Feijen J. Cell surface characteristics of coagulase-negative staphylococci and their adherence to fluorinated poly(ethylenepropylene). Infect Immun. 1986 Jan;51(1):294–301. doi: 10.1128/iai.51.1.294-301.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hussain M., Hastings J. G., White P. J. A chemically defined medium for slime production by coagulase-negative staphylococci. J Med Microbiol. 1991 Mar;34(3):143–147. doi: 10.1099/00222615-34-3-143. [DOI] [PubMed] [Google Scholar]
  12. Jensen E. T., Kharazmi A., Lam K., Costerton J. W., Høiby N. Human polymorphonuclear leukocyte response to Pseudomonas aeruginosa grown in biofilms. Infect Immun. 1990 Jul;58(7):2383–2385. doi: 10.1128/iai.58.7.2383-2385.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Lam J. S., Mutharia L. M., Hancock R. E., Høiby N., Lam K., Baek L., Costerton J. W. Immunogenicity of Pseudomonas aeruginosa outer membrane antigens examined by crossed immunoelectrophoresis. Infect Immun. 1983 Oct;42(1):88–98. doi: 10.1128/iai.42.1.88-98.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lambert P. A., Shorrock P. J., Aitchison E. J., Domingue P. A., Power M. E., Costerton J. W. Effect of in vivo growth conditions upon expression of surface protein antigens in Enterococcus faecalis. FEMS Microbiol Immunol. 1990 May;2(1):51–54. doi: 10.1111/j.1574-6968.1990.tb03478.x. [DOI] [PubMed] [Google Scholar]
  16. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  17. Patrick C. C., Plaunt M. R., Sweet S. M., Patrick G. S. Defining Staphylococcus epidermidis cell wall proteins. J Clin Microbiol. 1990 Dec;28(12):2757–2760. doi: 10.1128/jcm.28.12.2757-2760.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Smith D. G., Wilcox M. H., Williams P., Finch R. G., Denyer S. P. Characterization of cell envelope proteins of Staphylococcus epidermidis cultured in human peritoneal dialysate. Infect Immun. 1991 Feb;59(2):617–624. doi: 10.1128/iai.59.2.617-624.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Thomson-Carter F. M., Pennington T. H. Characterization of coagulase-negative staphylococci by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analyses. J Clin Microbiol. 1989 Oct;27(10):2199–2203. doi: 10.1128/jcm.27.10.2199-2203.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tojo M., Yamashita N., Goldmann D. A., Pier G. B. Isolation and characterization of a capsular polysaccharide adhesin from Staphylococcus epidermidis. J Infect Dis. 1988 Apr;157(4):713–722. doi: 10.1093/infdis/157.4.713. [DOI] [PubMed] [Google Scholar]
  21. Ward K. H., Olson M. E., Lam K., Costerton J. W. Mechanism of persistent infection associated with peritoneal implants. J Med Microbiol. 1992 Jun;36(6):406–413. doi: 10.1099/00222615-36-6-406. [DOI] [PubMed] [Google Scholar]
  22. Wilcox M. H., Smith D. G., Evans J. A., Denyer S. P., Finch R. G., Williams P. Influence of carbon dioxide on growth and antibiotic susceptibility of coagulase-negative staphylococci cultured in human peritoneal dialysate. J Clin Microbiol. 1990 Oct;28(10):2183–2186. doi: 10.1128/jcm.28.10.2183-2186.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wilcox M. H., Williams P., Smith D. G., Modun B., Finch R. G., Denyer S. P. Variation in the expression of cell envelope proteins of coagulase-negative staphylococci cultured under iron-restricted conditions in human peritoneal dialysate. J Gen Microbiol. 1991 Nov;137(11):2561–2570. doi: 10.1099/00221287-137-11-2561. [DOI] [PubMed] [Google Scholar]
  24. Williams P. Role of the cell envelope in bacterial adaptation to growth in vivo in infections. Biochimie. 1988 Aug;70(8):987–1011. doi: 10.1016/0300-9084(88)90263-5. [DOI] [PubMed] [Google Scholar]
  25. Younger J. J., Christensen G. D., Bartley D. L., Simmons J. C., Barrett F. F. Coagulase-negative staphylococci isolated from cerebrospinal fluid shunts: importance of slime production, species identification, and shunt removal to clinical outcome. J Infect Dis. 1987 Oct;156(4):548–554. doi: 10.1093/infdis/156.4.548. [DOI] [PubMed] [Google Scholar]

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