title: Binding and invasion of HeLa and MRC-5 cells by Streptococcus agalactiae.
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.
authors.affiliation: Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta, CanadaT6G 2J23.|Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta, CanadaT6G 2J22.|National Centre for Streptococcus, Provincial Laboratory for Public Health (Microbiology), Edmonton, Alberta, CanadaT6G 2J21.
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Tyrrell
authors.affiliation: Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta, CanadaT6G 2J22.|National Centre for Streptococcus, Provincial Laboratory for Public Health (Microbiology), Edmonton, Alberta, CanadaT6G 2J21.
authors.forename: Alexander
authors.initials: A
authors.lastname: Kennedy
authors.affiliation: National Centre for Streptococcus, Provincial Laboratory for Public Health (Microbiology), Edmonton, Alberta, CanadaT6G 2J21.
authors.forename: Sandra E
authors.initials: SE
authors.lastname: Shokoples
authors.affiliation: Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta, CanadaT6G 2J23.
authors.forename: Richard K
authors.initials: RK
authors.lastname: Sherburne
mesh_terms: D000199:Actins / Q000378:metabolism; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001422:Bacterial Adhesion; D002460:Cell Line; D015638:Cytochalasin D; D005347:Fibroblasts / Q000382:microbiology*; D005353:Fibronectins / Q000378:metabolism; D006367:HeLa Cells / Q000382:microbiology*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008855:Microscopy, Electron, Scanning; D008875:Middle Aged; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D013292:Streptococcus agalactiae / Q000378:metabolism* / Q000472:pathogenicity* / Q000502:physiology
chemical_list: D000199:Actins; D005353:Fibronectins; D015638:Cytochalasin D; D019869:Phosphatidylinositol 3-Kinases