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Infection and Immunity, November 1999, p. 5775-5783, Vol. 67, No. 11
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Outer Membrane Protein A-Promoted Actin
Condensation of Brain Microvascular Endothelial Cells Is Required
for Escherichia coli Invasion
Nemani V.
Prasadarao,1,2,*
Carol
A.
Wass,1
Monique F.
Stins,1
Hiroyuki
Shimada,2,3 and
Kwang Sik
Kim1,2
Division of Infectious
Diseases1 and Department of
Pathology,3 Childrens Hospital Los Angeles,
and University of Southern California School of
Medicine,2 Los Angeles, California 90027
Received 20 May 1999/Returned for modification 29 July
1999/Accepted 26 August 1999
Escherichia coli is the most common gram-negative
bacterium that causes meningitis during the neonatal period. We have
previously shown that the entry of circulating E. coli
organisms into the central nervous system is due to their ability to
invade the blood-brain barrier, which is composed of a layer of brain
microvascular endothelial cells (BMEC). In this report, we show by
transmission electron microscopy that E. coli transmigrates
through BMEC in an enclosed vacuole without intracellular
multiplication. The microfilament-disrupting agents cytochalasin D and
latrunculin A completely blocked E. coli invasion of BMEC.
Cells treated with the microtubule inhibitors nocodazole, colchicine,
vincristin, and vinblastine and the microtubule-stabilizing agent taxol
also exhibited 50 to 60% inhibition of E. coli invasion. Confocal laser scanning fluorescence microscopy showed F-actin condensation associated with the invasive E. coli but no
alterations in microtubule distribution. These results suggest that
E. coli uses a microfilament-dependent phagocytosis-like
endocytic mechanism for invasion of BMEC. Previously we showed that
OmpA expression significantly enhances the E. coli
invasion of BMEC. We therefore examined whether OmpA expression is
related to the recruitment of F-actin. OmpA+ E. coli induced the accumulation of actin in BMEC to a level similar
to that induced by the parental strain, whereas OmpA
E. coli did not. Despite the presence of OmpA, a
noninvasive E. coli isolate, however, did not show F-actin
condensation. OmpA+-E. coli-associated
condensation of F-actin was blocked by synthetic peptides corresponding
to the N-terminal extracellular domains of OmpA as well as BMEC
receptor analogues for OmpA, chitooligomers (GlcNAc
1-4GlcNAc
oligomers). These findings suggest that OmpA interaction is critical
for the expression or modulation of other bacterial proteins that will
subsequently cause actin accumulation for the uptake of bacteria.
*
Corresponding author. Mailing address: Division of
Infectious Diseases, Mail Stop no. 51, Childrens Hospital Los Angeles, 4650 Sunset Blvd., Los Angeles, CA 90027. Phone: (323) 669-5622. Fax:
(323) 660-2661. E-mail: pnemani{at}chla.usc.edu.
Infection and Immunity, November 1999, p. 5775-5783, Vol. 67, No. 11
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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