Transcriptional activation of nitric oxide synthase-2, and NO-induced cell death, in mouse cerebrovascular endothelium exposed to Neisseria meningitidis.
The site and mechanisms by which meningococci gain access to the CNS are unclear. In this study we determined whether production of nitric oxide (NO) is part of the host (endothelial cell) response to meningococcal cell lysate, and the consequences for endothelial cell viability. Expression of NO synthase type II (NOS-2) mRNA, protein and enzyme activity were investigated in mouse cerebrovascular endothelial cells exposed to sonicated Neisseria meningitidis. The production of nitrite peaked after 48 h of incubation, and this reflected transcriptional activation of the NOS-2 gene and increased expression of the NOS-2 protein. This endothelial response was independent of meningococcal lipopolysaccharide production. Endothelial cell death occurred as a result of NO production, and addition of a NOS inhibitor prevented cell death, but the cells did not exhibit features of apoptosis. However, inhibition of poly (ADP-ribose) polymerase (PARP) decreased the rate of cell death by more than 40%. These data indicate that N. meningitidis increases expression of NOS-2 in endothelial cells and causes cell death. Such an effect could contribute to meningococcal entry into the CNS in situ.
Constantin
Despina
D
Institute of Cell Signalling, Division of Microbiology and Infectious Diseases, Medical School, Queen's Medical Centre, University of Nottingham, UK.
Ala'Aldeent
Dlawer
D
Murphy
Sean
S
D000818:Animals; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000502:physiology; D002560:Cerebrovascular Circulation; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism* / Q000382:microbiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D009345:Neisseria meningitidis / Q000737:chemistry; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000235:genetics / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D009573:Nitrites / Q000378:metabolism; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D012333:RNA, Messenger / Q000096:biosynthesis; D013045:Species Specificity; D013347:Subcellular Fractions / Q000737:chemistry
D004791:Enzyme Inhibitors; D008070:Lipopolysaccharides; D009573:Nitrites; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D012333:RNA, Messenger; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse