title: Disease and Carrier Isolates of Neisseria meningitidis Cause G1 Cell Cycle Arrest in Human Epithelial Cells.
abstract: Microbial pathogens have developed several mechanisms to modulate and interfere with host cell cycle progression. In this study, we analyzed the effect of the human pathogen Neisseria meningitidis on the cell cycle of epithelial cells. Two pathogenic isolates, as well as two carrier isolates, were tested for their ability to adhere to and invade into the epithelial cell lines Detroit 562 and NP69 and to modulate the cell cycle. We found that all isolates adhered equally well to both Detroit 562 and NP69 cells, whereas the carrier isolates were significantly less invasive. Using propidium iodide staining and 5-ethynyl-2'-deoxyuridine pulse-labeling, we provide evidence that meningococcal infection arrested cells in the G1 phase of the cell cycle at 24 h postinfection. In parallel, a significant decrease of cells in the S phase was observed. Interestingly, G1-phase arrest was only induced after infection with live bacteria but not with heat-killed bacteria. By Western blotting we demonstrate that bacterial infection resulted in a decreased protein level of the cell cycle regulator cyclin D1, whereas cyclin E expression levels were increased. Furthermore, N. meningitidis infection induced an accumulation of the cyclin-dependent kinase inhibitor (CKI) p21(WAF1/CIP1) that was accompanied by a redistribution of this CKI to the cell nucleus, as shown by immunofluorescence analysis. Moreover, the p27(CIP1) CKI was redistributed and showed punctate foci in infected cells. In summary, we present data that N. meningitidis can interfere with the processes of host cell cycle regulation.
authors.affiliation: Institute of Hygiene and Microbiology, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: von Papen
authors.affiliation: Institute of Hygiene and Microbiology, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Wilhelm F
authors.initials: WF
authors.lastname: Oosthuysen
authors.affiliation: Institute of Hygiene and Microbiology, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Becam
authors.affiliation: Institute of Hygiene and Microbiology, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Heike
authors.initials: H
authors.lastname: Claus
authors.affiliation: Institute of Hygiene and Microbiology, University of Wuerzburg, Wuerzburg, Germany aunkmeir@hygiene.uni-wuerzburg.de.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Schubert-Unkmeir
mesh_terms: D001422:Bacterial Adhesion / Q000502:physiology; D015153:Blotting, Western; D059447:Cell Cycle Checkpoints; D002467:Cell Nucleus / Q000378:metabolism; D019938:Cyclin D1 / Q000378:metabolism; D019927:Cyclin E / Q000378:metabolism; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D004847:Epithelial Cells / Q000382:microbiology* / Q000502:physiology; D054884:Host-Pathogen Interactions; D006801:Humans; D008589:Meningococcal Infections / Q000378:metabolism / Q000382:microbiology*; D009345:Neisseria meningitidis / Q000378:metabolism / Q000502:physiology*
chemical_list: D019927:Cyclin E; D050759:Cyclin-Dependent Kinase Inhibitor p21; D019938:Cyclin D1