Enterotoxigenic Escherichia coli (ETEC) is one of the major causes of childhood morbidity and mortality in developing countries. In 2010, annual mortality from illness due to ETEC was estimated at 157,000 deaths (9 percent of all deaths attributed to diarrhea) and approximately 1 percent of all deaths in children 28 days to 5 years of age. ETEC cause disease by colonizing the small intestine with colonization factor antigens and cause secretory diarrhea through the actions of heat-labile (LT) or heat-stable (ST) enterotoxins. Development of natural immunity to ETEC requires years of recurrent infections, which raises the question as to what about this organism prevents the development of immunity. LT is highly immunogenic and induces antibodies to itself when given to human volunteers. On the other hand ST is a small non-immunogenic secreted peptide. We have recently begun to investigate the contribution of ST on ETEC pathogenesis. Our preliminary data indicate (1) ST expression is higher in defined 4AA media as compared to LB, sta1 gene (encoding STh) is up-regulated under anaerobic conditions, as could be found within the host intestinal tract, and iron-depletion decreases ST expression (2) ST binds both iron and iron- sulfur (FeS) clusters under these anaerobic conditions, in vitro (3) iron-bound forms of ST elicit less secretion than native ST in an in vivo model and less cGMP from intestinal epithelial cells in cell culture than native ST, and (4) ST suppresses fecal IgA and Th17 immune responses against ETEC H10407 (whole cell and O78- LPS). Based on the preliminary data, the hypotheses are that (1) sta1 is up-regulated in response to anaerobic conditions by specific cis and trans factors (2) ST can bind iron and FeS clusters, which regulate its toxicity, and (3) ST suppresses mucosal immunity to ETEC antigens thereby maintaining susceptibility of the host to recurrent infections by the same organism mediated by iron sensing.
The specific aims are 1) Determine the environmental conditions that regulate transcription of genes encoding ST, 2) Characterize the iron and iron- sulfur cluster binding properties of ST and how this controls ST enterotoxicity, and 3) Determine how ST modifies mucosal immune responses to ETEC antigens. At the conclusion of these studies, growth conditions that result in enhanced ST expression and secretion will be elucidated. In addition, the role of iron and FeS clusters on ST toxicity will be clarified. Lastly, mechanisms concerning ST-mediated suppression of mucosal immune responses to ETEC antigens will provide compelling evidence for why it takes years of recurrent ETEC infections to develop natural immunity and inform vaccine development.
Enterotoxigenic Escherichia coli (ETEC) is one of the top four pathogens causing moderate to severe diarrhea in children aged 0-60 months. ETEC is a significant global health threat causing mortality in over 150,000 children per year. While there is no licensed ETEC vaccine, this project is designed to elucidate the role of the heat-stable enterotoxin (ST) in ETEC pathogenesis to inform vaccine development.
