Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 100, Issue 14, Pages 8508-8513Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1431769100
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Funding
- FIC NIH HHS [D43 TW005572, D43 TW05572] Funding Source: Medline
- NIAID NIH HHS [R01 AI44487, R01 AI040725, R01 AI25096, R01 AI40725, R01 AI025096] Funding Source: Medline
- NICHD NIH HHS [U01 HD039165, UO1 HD39165] Funding Source: Medline
- NIDCR NIH HHS [R01 DE013523, R01 DE13523] Funding Source: Medline
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In vivo-induced antigen technology is a method to identify proteins expressed by pathogenic bacteria during human infection. Sera from 10 patients convalescing from cholera infection in Bangladesh were pooled, adsorbed against in vitro-grown El Tor Vibrio cholerae 01, and used to probe a genomic expression library in Escherichia coli constructed from El Tor V. cholerae 01 strain N16961. We identified 38 positive clones in the screen, encoding pili (PilA and TcpA), cell membrane proteins (PilQ, MshO, MshP, and CapK), methyl-accepting chemotaxis proteins, chemotaxis and motility proteins (CheA and CheR), a quorum-sensing protein (LuxP), and four hypothetical proteins. Analysis of immune responses to purified PilA and TcpA in individual patients demonstrated that the majority seroconverted to these proteins, confirming results with pooled sera. These results suggest that PilA and its outer membrane secretin, PilQ, are expressed during human infection and may be involved in colonization of the gastrointestinal tract. These results also demonstrate substantial immune responses to TcpA in patients infected with El Tor V. cholerae O1. In vivo-induced antigen technology provides a simple method for identifying microbial proteins expressed during human infection, but not during in vitro growth.
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