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Infection and Immunity, November 1998, p. 5260-5267, Vol. 66, No. 11
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Interaction of Listeria monocytogenes
with Human Brain Microvascular Endothelial Cells: InlB-Dependent
Invasion, Long-Term Intracellular Growth, and Spread from Macrophages
to Endothelial Cells
Lars
Greiffenberg,1
Werner
Goebel,1
Kwang Sik
Kim,2,3
Inge
Weiglein,1
Andreas
Bubert,1
Fredi
Engelbrecht,1
Monique
Stins,2 and
Michael
Kuhn1,*
Lehrstuhl für Mikrobiologie,
Theodor-Boveri-Institut für Biowissenschaften der
Universität Würzburg, 97074 Würzburg,
Germany,1 and
Division of Infectious
Diseases, Childrens Hospital Los Angeles,2 and
Departments of Pediatrics, Molecular Microbiology, and
Immunology, USC School of Medicine,3 Los
Angeles, California 90027
Received 8 April 1998/Returned for modification 17 June
1998/Accepted 6 August 1998
Invasion of endothelial tissues may be crucial in a Listeria
monocytogenes infection leading to meningitis and/or
encephalitis. Internalization of L. monocytogenes into
endothelial cells has been previously demonstrated by using human
umbilical vein endothelial cells as a model system. However, during the
crossing of the blood-brain barrier, L. monocytogenes most
likely encounters brain microvascular endothelial cells which are
strikingly different from macrovascular or umbilical vein endothelial
cells. In the present study human brain microvascular endothelial cells
(HBMEC) were used to study the interaction of L. monocytogenes with endothelial cells, which closely resemble
native microvascular endothelial cells of the brain. We show that
L. monocytogenes invades HBMEC in an InlB-dependent and
wortmannin-insensitive manner. Once within the HBMEC, L. monocytogenes replicates efficiently over a period of at least
18 h, moves intracellularly by inducing actin tail formation, and
spreads from cell to cell. Using a green fluorescent protein-expressing
L. monocytogenes strain, we present direct evidence that
HBMEC are highly resistant to damage by intracellularly growing
L. monocytogenes. Infection of HBMEC with L. monocytogenes results in foci of heavily infected, but largely
undamaged endothelial cells. Heterologous plaque assays with L. monocytogenes-infected P388D1 macrophages as vectors
demonstrate efficient spreading of L. monocytogenes into
HBMEC, fibroblasts, hepatocytes, and epithelial cells, and this
phenomenon is independent of the inlC gene product.
*
Corresponding author. Mailing address: Lehrstuhl
für Mikrobiologie, Theodor-Boveri-Institut für
Biowissenschaften der Universität Würzburg, Am Hubland,
97074, Würzburg, Germany. Phone: (49)-931-8884421. Fax:
(49)-931-8884402. E-mail:
kuhn{at}biozentrum.uni-wuerzburg.de.
Infection and Immunity, November 1998, p. 5260-5267, Vol. 66, No. 11
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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