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Transcriptional Profiling of Vibrio cholerae Recovered Directly from Patient Specimens during Early and Late Stages of Human Infection

Division of Infectious Diseases, Massachusetts General Hospital,¹ Department of Medicine,² Department of Microbiology and Molecular Genetics, Harvard Medical School,³ Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston,⁶ Department of Biostatistics, Harvard University, Cambridge, Massachusetts,⁴ International Centre for Diarrhoeal Disease Research, Dhaka, Bangladesh⁵

Received 17 November 2004/ Returned for modification 16 January 2005/ Accepted 15 March 2005

Understanding gene expression by bacteria during the actual course of human infection may provide important insights into microbial pathogenesis. In this study, we evaluated the transcriptional profile of Vibrio cholerae, the causative agent of cholera, in clinical specimens from cholera patients. We collected samples of human stool and vomitus that were positive by dark-field microscopy for abundant vibrios and used a microarray to compare gene expression in organisms recovered directly from specimens collected during the early and late stages of human infection. Our results reveal that V. cholerae gene expression within the human host environment differs from patterns defined in in vitro models of pathogenesis. tcpA, the major subunit of the essential V. cholerae colonization factor, was significantly more highly expressed in early than in late stages of infection; however, the genes encoding cholera toxin were not highly expressed in either phase of human infection. Furthermore, expression of the virulence regulators toxRS and tcpPH was uncoupled. Interestingly, the pattern of gene expression indicates that the human upper intestine may be a uniquely suitable environment for the transfer of genetic elements that are important in the evolution of pathogenic strains of V. cholerae. These findings provide a more detailed assessment of the transcriptome of V. cholerae in the human host than previous studies of organisms in stool alone and have implications for cholera control and the design of improved vaccines.

Editor: W. A. Petri, Jr.

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