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Infect Immun, June 1998, p. 2410-2419, Vol. 66, No. 6
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Cytochalasin-Induced Actin Disruption of Polarized Enterocytes Can Augment Internalization of Bacteria

Carol L. Wells,1,2,* Elisabeth M. A. van de Westerlo,1 Robert P. Jechorek,1 Holly M. Haines,1 and Stanley L. Erlandsen3

Departments of Laboratory Medicine and Pathology,1 Surgery,2 and Cell Biology and Neuroanatomy,3 University of Minnesota, Minneapolis, Minnesota 55455-0385

Received 7 January 1998/Returned for modification 20 February 1998/Accepted 4 March 1998

Cytochalasin-induced actin disruption has often been associated with decreased bacterial internalization by cultured epithelial cells, although polarized enterocytes have not been systematically studied. In assays using confluent polarized HT-29 enterocytes, cytochalasin D appeared to increase internalization of wild-type Salmonella typhimurium, Proteus mirabilis, and Escherichia coli. HeLa and HEp-2 epithelial cells, as well as HT-29 and Caco-2 enterocytes, were used to clarify this unexpected observation. Resulting data showed that cytochalasin D was associated with increased internalization of S. typhimurium and P. mirabilis by both HT-29 and Caco-2 enterocytes and with increased internalization of E. coli by HT-29 enterocytes; with either HeLa or HEp-2 cells, cytochalasin was associated with no change or a decrease in internalization of these same bacterial strains. Cytochalasin caused decreased internalization of Listeria monocytogenes by HT-29, Caco-2, HeLa, and HEp-2 cells, indicating that cytochalasin did not consistently augment bacterial internalization by polarized enterocytes. Fluorescein-labeled phalloidin confirmed marked disruption of filamentous actin in cytochalasin-treated HT-29, Caco-2, HeLa, and HEp-2 cells. Cytochalasin had no noticeable effect on epithelial viability but caused distorted apical microvilli, cell rounding, and separation of adjacent enterocytes in confluent cultures (with a corresponding decrease in transepithelial electrical resistance). Scanning electron microscopy showed that cytochalasin-induced enhanced bacterial internalization was associated with preferential bacterial adherence on the exposed enterocyte lateral surface. Colchicine, used to disrupt microtubules, had no noticeable effect on bacterial internalization by HT-29 or Caco-2 enterocytes. These data indicated that for HT-29 and Caco-2 enterocytes, cytochalasin-induced disruption of filamentous actin might augment internalization of some bacterial species by a mechanism that appeared to involve exposure of the enterocyte lateral surface.

* Corresponding author. Mailing address: Department of Laboratory Medicine and Pathology, Box 609 UMHC, 420 Delaware St., SE, University of Minnesota, Minneapolis, MN 55455-0385. Phone: (612) 625-5951. Fax: (612) 625-5622. E-mail: wells002{at}maroon.tc.umn.edu.

Infect Immun, June 1998, p. 2410-2419, Vol. 66, No. 6
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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