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Infection and Immunity, October 2001, p. 6310-6317, Vol. 69, No. 10
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.10.6310-6317.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Vibrio cholerae-Induced Cellular
Responses of Polarized T84 Intestinal Epithelial Cells Are Dependent on
Production of Cholera Toxin and the RTX Toxin
Karla Jean
Fullner,1,*
Wayne I.
Lencer,2,3 and
John
J.
Mekalanos1,2
Department of Microbiology and Molecular
Genetics1 and the Harvard Digestive
Diseases Center,2 Harvard Medical School, and
Pediatric Gastroenterology, Children's
Hospital,3 Boston, Massachusetts
Received 20 April 2001/Returned for modification 12 June
2001/Accepted 12 July 2001
To study the utility of in vitro-polarized intestinal cell
monolayers for modeling Vibrio cholerae-host cell
interactions, we added live V. cholerae bacteria to the
apical surfaces of polarized T84 cell monolayers and monitored changes
in electrical properties. We found that both classical and El Tor
strains produce cholera toxin after addition to the monolayer, but
induction is most likely due to medium components rather than
bacterium-cell interactions. We also found that the RTX toxin is
produced by El Tor strains. This toxin caused a loss of the barrier
function of the paracellular tight junction that was measured as a
decrease in transepithelial resistance. This decrease occurred when
bacteria were added to either the apical or basolateral surfaces,
indicating that the RTX toxin receptor is expressed on both surfaces.
These results are discussed with regard to the applicability of the
polarized T84 cell monolayers as an in vitro model of host-pathogen interactions.
*
Corresponding author. Present address: Department of
Microbiology-Immunology, 303 E. Chicago Ave., Morton 6-626,
Northwestern University Medical School, Chicago, IL 60611. Phone: (312)
503-2162. Fax: (312) 503-1339. E-mail:
k-fullner{at}northwestern.edu.
Infection and Immunity, October 2001, p. 6310-6317, Vol. 69, No. 10
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.10.6310-6317.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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