Biological nanofactories target and activate epithelial cell surfaces for modulating bacterial quorum sensing and interspecies signaling

ACS Nano. 2010 Nov 23;4(11):6923-31. doi: 10.1021/nn1013066. Epub 2010 Oct 28.

Abstract

In order to control the behavior of bacteria present at the surface of human epithelial cells, we have created a biological "nanofactory" construct that "coats" the epithelial cells and "activates" the surface to produce the bacterial quorum sensing signaling molecule, autoinducer-2 (AI-2). Specifically, we demonstrate directed modulation of signaling among Escherichia coli cells grown over the surface of human epithelial (Caco-2) cells through site-directed attachment of biological nanofactories. These "factories" comprise a fusion protein expressed and purified from E. coli containing two AI-2 bacterial synthases (Pfs and LuxS), a protein G IgG binding domain, and affinity ligands for purification. The final factory is fabricated ex vivo by incubating with an anti-CD26 antibody that binds the fusion protein and specifically targets the CD26 dipeptidyl peptidase found on the outer surface of Caco-2 cells. This is the first report of the intentional "in vitro" synthesis of bacterial autoinducers at the surface of epithelial cells for the redirection of quorum sensing behaviors of bacteria. We envision tools such as this will be useful for interrogating, interpreting, and disrupting signaling events associated with the microbiome localized in human intestine and other environments.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Caco-2 Cells
  • Coculture Techniques
  • Dipeptidyl Peptidase 4 / immunology
  • Epithelial Cells / cytology*
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism*
  • Escherichia coli / cytology*
  • Escherichia coli / drug effects
  • Escherichia coli / metabolism
  • Homoserine / analogs & derivatives
  • Homoserine / biosynthesis
  • Humans
  • Immunoglobulin G / immunology
  • Immunoglobulin G / metabolism
  • Lactones
  • Nanotechnology / methods*
  • Quorum Sensing / drug effects*
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Fusion Proteins / pharmacology
  • Vibrio / cytology*
  • Vibrio / drug effects
  • Vibrio / metabolism

Substances

  • Immunoglobulin G
  • Lactones
  • N-octanoylhomoserine lactone
  • Recombinant Fusion Proteins
  • Homoserine
  • Dipeptidyl Peptidase 4