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Infection and Immunity, July 2004, p. 4240-4248, Vol. 72, No. 7
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.7.4240-4248.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Transcriptional Responses of Intestinal Epithelial Cells to Infection with Vibrio cholerae

Neil R. Stokes,1,2,{dagger} Xin Zhou,1,2 Stephen J. Meltzer,3,4 and James B. Kaper1,2*

Center for Vaccine Development,1 Department of Microbiology and Immunology,2 Division of Gastroenterology, Department of Medicine,3 Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, Maryland4

Received 1 February 2004/ Accepted 23 March 2004

Vibrio cholerae is a noninvasive enteric bacterium that causes the severe diarrheal disease cholera. Candidate cholera vaccines have been engineered by deleting genes encoding known virulence factors in V. cholerae; however, many of these attenuated strains were still reactogenic in human volunteers. In this study, DNA arrays were utilized to monitor the transcriptional responses of human intestinal epithelial cells (T84) to eight strains of V. cholerae, including attenuated, toxigenic, and environmental isolates. cDNA probes generated from host RNA samples were hybridized against low- and high-density gene arrays. V. cholerae induced the transcription of a variety of host genes and repressed the expression of a lower number of genes. Expression patterns were confirmed for certain genes by reverse transcriptase PCR and enzyme-linked immunosorbent assays. A core subset of genes was found to be differentially regulated in all experiments. These genes included genes involved in innate mucosal immunity, intracellular signaling, and cellular proliferation. Reactogenic vaccine strains induced greater expression of genes for certain proinflammatory cytokines than nonreactogenic strains. Wild-type and attenuated derivatives induced and repressed many genes in common, although there were differences in the transcription profiles. These results indicate that the types of host genes modulated by attenuated V. cholerae, and the extent of their induction, may mediate the symptoms seen with reactogenic cholera vaccine strains.

* Corresponding author. Mailing address: Center for Vaccine Development, University of Maryland School of Medicine, 685 West Baltimore St., Baltimore, MD 21201. Phone: (410) 706-2344. Fax: (410) 706-0182. E-mail: jkaper{at}umaryland.edu.

Editor: V. J. DiRita

{dagger} Present address: Prolysis Limited, Yarnton, Oxfordshire OX5 1PF, United Kingdom.

Infection and Immunity, July 2004, p. 4240-4248, Vol. 72, No. 7
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.7.4240-4248.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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