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Volume 148, Issue 12

Binding and invasion of HeLa and MRC-5 cells by Free

Abstract

The interactions of group B streptococci (GBS) with HeLa cells (an epithelial cell line) and MRC-5 cells (a fibroblastic cell line) were explored. A host-cell invasion assay using GBS strains from all serotypes revealed that GBS invaded HeLa cells to a greater extent than MRC-5 cells. One strain, a serotype V (NCS13), was highly invasive against HeLa cells. All strains were poorly invasive against MRC-5 cells. Further characterization of the binding of NCS13 to HeLa and MRC-5 cell surfaces showed that the lack of recoverable c.f.u. from MRC-5 cells was due to a lack of binding of NCS13 to the MRC-5 cell surface in comparison to HeLa cells. Although fibronectin had been reported to bind to GBS, fibronectin assays showed 2·7-fold more fibronectin on the MRC-5 cell surface in comparison to HeLa cells, suggesting that other extracellular matrix proteins besides fibronectin may be involved in GBS binding. Scanning electron microscopy of NCS13 and HeLa cells over a 6 h time period showed increased numbers of NCS13 on the HeLa cell surface over time until cell death at 6 h. Direct contact of the HeLa cell surface by NCS13 was found to be necessary for cell death to occur. Further scanning electron microscopy studies found that, once GBS are bound to the HeLa cell surface, HeLa cell microvilli entwine the bacteria, which then enter the HeLa cell in a polar fashion. Cytoskeletal actin is involved, as this process is disrupted by cytochalasin D, and recruitment of actin is visible at the site of adherent chains of GBS. Also, the host-cell signalling enzyme, PI 3-kinase, is involved in the GBS internalization process, since the PI 3-kinase inhibitor, wortmannin, inhibited NCS13 invasion of HeLa cells in a dose-dependent manner.

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Keyword(s): actin , GBS, group B streptococci , NCS, National Centre for Streptococcus , phosphatidylinositol 3-kinase , SBA, sheep blood agar , scanning electron microscopy , SEM, scanning electron microscopy , streptococci and TEM, transmission electron microscopy
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2002-12-01
2024-04-25
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References

  1. Allardice J. G., Baskett T. F., Seshia M. M., Bowman N., Malazdrewicz R. 1982; Perinatal group B streptococcal colonization and infection. Am J Obstet Gynecol 142:617–620
     [Google Scholar]
  2. Anthony B., Eisenstadt R., Carter J., Kim K., Hobel C. 1981; Genital and intestinal carriage of group B streptococci during pregnancy. J Infect Dis 143:761–766 [CrossRef]
     [Google Scholar]
  3. Beckmann C., Waggoner J. D., Harris T. O., Tamura G. S., Rubens C. E. 2002; Identification of novel adhesins from group B streptococci by use of phage display reveals that C5a peptidase mediates fibronectin binding. Infect Immun 70:2869–2876 [CrossRef]
     [Google Scholar]
  4. Campbell J. R., Hillier S. L., Krohn M. A., Ferrieri P., Zaleznik D. F., Baker C. J. 2000; Group B streptococcal colonization and serotype-specific immunity in pregnant women at delivery. Obstet Gynecol 96:498–503 [CrossRef]
     [Google Scholar]
  5. Cheng Q., Stafslien S., Purushothaman S. P., Cleary P. 2002; The group B streptococcal C5a peptidase is both a specific protease and an invasin. Infect Immun 70:2408–2413 [CrossRef]
     [Google Scholar]
  6. Edwards M., Baker C. 1995; Streptococcus agalactiae (Group B Streptococcus ). In Principles and Practices of Infectious Diseases pp 2156–2167 Edited by Mandell G. Philadelphia,PA: Churchill Livingstone;
     [Google Scholar]
  7. Farley M., Harvey R., Stull T., Smith J., Schuchat A., Wenger J., Stephens D. 1993; A population-based assessment of invasive disease due to group B Streptococcus in nonpregnant adults. N Engl J Med 328:1807–1811 [CrossRef]
     [Google Scholar]
  8. Gibson R., Lee M., Soderland C., Chi E. Y., Rubens C. E. 1993; Group B streptococci invade endothelial cells: type III capsular polysaccharide attenuates invasion. Infect Immun 61:478–485
     [Google Scholar]
  9. Hulse M., Smith S., Chi E. Y., Pham A., Rubens C. E. 1993; Effect of type III group B streptococcal capsular polysaccharide on invasion of respiratory epithelial cells. Infect Immun 61:4835–4841
     [Google Scholar]
  10. Kallman J., Kihlstrom E. 1997; Penetration of Group B streptococci through polarized Madin–Darby canine kidney cells. Pediatr Res 42:799–804 [CrossRef]
     [Google Scholar]
  11. Knutton S., Baldwin T., Williams P. H., McNeish A. S. 1989; Actin accumulation at sites of bacterial adhesion to tissue culture cells: basis of a new diagnostic test for enteropathogenic and enterohemorrhagic Escherichia coli . Infect Immun 57:1290–1298
     [Google Scholar]
  12. Lalonde M., Segura M., Lacoutre S., Gottschalk M. 2000; Interactions between Streptococcus suis serotype 2 and different epithelial cell lines. Microbiology 146:1913–1921
     [Google Scholar]
  13. Marchlewicz B. A., Duncan J. L. 1980; Properties of a hemolysin produced by group B streptococci. Infect Immun 30:805–813
     [Google Scholar]
  14. Nizet V., Gibson R., Chi E. Y., Framson P., Hulse M., Rubens C. E. 1996; Group B streptococcal β-hemolysin expression is associated with injury of lung epithelial cells. Infect Immun 64:3818–3826
     [Google Scholar]
  15. Nizet V., Kim K. S., Stins M., Jonas M., Chi E. Y., Nguyen D., Rubens C. E. 1997; Invasion of brain microvascular endothelial cells by group B streptococci. Infect Immun 65:5074–5081
     [Google Scholar]
  16. Ozeri V., Rosenshine I., Ben-ZeEv A., Bokoch G. M., Jou T., Hanski E. 2001; De novo formation of focal complex-like structures in host cells by invading streptococci. Mol Microbiol 41:561–573 [CrossRef]
     [Google Scholar]
  17. Perch B., Kjems E., Henrichsen J. 1979; New seroytpes of group B streptococci isolated from human sources. J Clin Microbiol 10:109–110
     [Google Scholar]
  18. Platt M. W. 1995; In vivo hemolytic activity of group B Streptococcus is dependent on erythrocyte–bacteria contact and independent of a carrier molecule. Curr Microbiol 31:5–9 [CrossRef]
     [Google Scholar]
  19. Rubens C. E., Raff H. V., Jackson J. C., Chi E. Y., Bielitzki J. T., Hillier S. L. 1991; Pathophysiology and histopathology of group B streptococcal sepsis in Macaca nemestrina primates induced after intraamniotic inoculation: evidence for bacterial cellular invasion. J Infect Dis 164:320–330 [CrossRef]
     [Google Scholar]
  20. Rubens C. E., Smith S., Hulse M., Chi E. Y., van Belle G. 1992; Respiratory epithelial cell invasion by group B streptococci. Infect Immun 60:5157–5163
     [Google Scholar]
  21. Schuchat A., Oxtoby M., Cochi S., Sikes R. K., Hightower A., Plikaytis B. D., Broome C. V. 1990; Population-based risk factors for neonatal group B streptococcal disease: results of a cohort study in metropolitan Atlanta. J Infect Dis 162:672–677 [CrossRef]
     [Google Scholar]
  22. Schuchat A., Deaver-Robinson K., Plikaytis B. D., Zangwell K. M., Mohle-Boetani J., Wenger J. D. 1994; Multistate case-control study of maternal risk factors for neonatal group B streptococcal disease. The Active Surveillance Study Group. Pediatr Infect Dis J 13:623–629 [CrossRef]
     [Google Scholar]
  23. Spellerberg B., Rozdzinski E., Martin S., Weber-Heynemann J., Schnitzler N., Lutticken R., Podbielski A. 1999; Lmb, a protein with similarities to the Lra1 adhesion family, mediates attachment of Streptococcus agalactiae to human laminin. Infect Immun 67:871–878
     [Google Scholar]
  24. Spellerberg B., Rozdzinski E., Martin S., Weber-Heyneman J., Luticken R. 2002; rgf encodes a novel two-component signal transduction system of Streptococcus agalactiae . Infect Immun 70:2434–2440 [CrossRef]
     [Google Scholar]
  25. Tamura G. S., Rubens C. E. 1994; Host–bacterial interactions in the pathogenesis of group B streptococcal infections. Curr Opin Infect Dis 7:317–322 [CrossRef]
     [Google Scholar]
  26. Tamura G. S., Rubens C. E. 1995; Group B streptococci adhere to a variant of fibronectin attached to a solid phase. Mol Microbiol 15:581–589 [CrossRef]
     [Google Scholar]
  27. Tamura G. S., Nittayajarn A. 2000; Group B streptococci and other Gram-positive cocci bind to cytokeratin 8. Infect Immun 68:2129–2134 [CrossRef]
     [Google Scholar]
  28. Tamura G. S., Nittayajain A., Schoentag D. L. 2002; A glutamine transport gene, glnQ , is required for fibronectin adherence and virulence of group B streptococci. Infect Immun 70:2877–2885 [CrossRef]
     [Google Scholar]
  29. Tyrrell G. J., Senzilet L. D., Spika J. S., Kertesz D. A., Alagaratnam M., Lovgren M., Talbot J. A. the Sentinel Health Unit Surveillance System Site Coordinators 2000; Invasive disease due to group B streptococcal infection in adults: results from a Canadian, population-based, active laboratory surveillance study–1996. J Infect Dis 182:168–173 [CrossRef]
     [Google Scholar]
  30. Valentin-Weigand P., Chhatwal G. S. 1995; Correlation of epithelial cell invasiveness of group B streptococci with clinical source of isolation. Microb Pathog 19:83–91 [CrossRef]
     [Google Scholar]
  31. Valentin-Weigand P., Benkel P., Rhode M., Chhatwal G. S. 1996; Entry and intracellular survival of group B streptococci in J774 macrophages. Infect Immun 64:2467–2473
     [Google Scholar]
  32. Valentin-Weigand P., Jungnitz H., Zock A., Rhode M., Chhatwal G. S. 1997; Characterization of group B streptococcal invasion in HEp-2 epithelial cells. FEMS Microbiol Lett 147:69–74 [CrossRef]
     [Google Scholar]
  33. Wagner M., Murai T., Wagner B., Gunther E., Jelinkova J. 1994; JM9 strains, a new type of group B streptococci from Japan. Zentbl Bakteriol 280:488–496 [CrossRef]
     [Google Scholar]
  34. Wilkinson H. W. 1997; Nontypable group B streptococci isolated from human sources. J Clin Microbiol 6:183–184
     [Google Scholar]
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Binding and invasion of HeLa and MRC-5 cells by Streptococcus agalactiae
Microbiology 148, 3921 (2002); https://doi.org/10.1099/00221287-148-12-3921
/content/journal/micro/10.1099/00221287-148-12-3921
/content/journal/micro/10.1099/00221287-148-12-3921
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