Outer membrane protein A expression in Enterobacter sakazakii is required to induce microtubule condensation in human brain microvascular endothelial cells for invasion.
Enterobacter sakazakii (ES) causes neonatal meningitis and necrotizing enterocolitis with case-fatality rates among infected infants ranging from 40 to 80%. Very little is known about the mechanisms by which these organisms cause disease. Here, we demonstrate that ES invades human brain microvascular endothelial cells (HBMEC) with higher frequency when compared with epithelial cells and endothelial cells from different origins. The entry of ES into HBMEC requires the expression of outer membrane protein A (OmpA), as the OmpA-deletion mutant was sevenfold less invasive than the wild type ES and the bacterium does not multiply inside HBMEC. Anti-OmpA antibodies generated against the OmpA of Escherichia coli K1, which also recognize the OmpA of ES, did not prevent the invasion of ES in HBMEC. ES invasion depends on microtubule condensation in HBMEC and is independent of actin filament reorganization. Both PI3-kinase and PKC-alpha were activated during ES entry into HBMEC between 15 min and 30 min of infection. Concomitantly, overexpression of dominant negative forms of PI3-kinase and PKC-alpha significantly inhibited the invasion of ES into HBMEC. In summary, ES invasion of HBMEC is dependent on the expression of OmpA similar to that of E. coli K1; however, the epitopes involved in the interaction with HBMEC appears to be different.
Singamsetty
Vijay K
VK
Division of Infectious Diseases, The Saban Research Institute, Childrens Hospital Los Angeles, Los Angeles, CA 90027, USA.
Wang
Ying
Y
Shimada
Hiroyuki
H
Prasadarao
Nemani V
NV
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D001921:Brain / Q000201:enzymology / Q000382:microbiology*; D002478:Cells, Cultured; D044083:Cronobacter sakazakii / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity*; D042783:Endothelial Cells / Q000378:metabolism / Q000382:microbiology; D004730:Endothelium, Vascular / Q000201:enzymology / Q000382:microbiology; D004756:Enterobacteriaceae Infections / Q000201:enzymology / Q000378:metabolism / Q000382:microbiology*; D004847:Epithelial Cells / Q000201:enzymology / Q000378:metabolism / Q000382:microbiology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008870:Microtubules / Q000201:enzymology / Q000378:metabolism* / Q000382:microbiology; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics / Q000378:metabolism; D051571:Protein Kinase C-alpha / Q000235:genetics / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D019869:Phosphatidylinositol 3-Kinases; D051571:Protein Kinase C-alpha; C078167:OMPA outer membrane proteins