Meta Kuehn (Primary)
Duke Box 3711, Durham, NC 27710
Research Drive, 220A Nanaline H Duke, Durham, NC 27710
Enterotoxigenic E. coli (ETEC) causes traveler's diarrhea and infant mortality in underdeveloped countries, and Pseudomonas aeruginosa is an opportunistic pathogen for immunocompromised patients. Like all gram negative bacteria studied to date, ETEC and P. aeruginosa produce small outer membrane vesicles that can serve as delivery "bombs" to host tissues. Vesicles contain a subset of outer membrane and soluble periplasmic proteins and lipids. In tissues and sera of infected hosts, vesicles have been observed to bud from the pathogen and come in close contact with epithelial cells. Despite their association with disease, the ability of pathogenic bacteria to distribute an arsenal of virulence factors to the host cells via vesicles remains relatively unexplored.
In our lab, we focus on the genetic, biochemical and functional features of bacterial vesicle production. Using a genetic screen, we have identified genes essential in the vesiculation process, we have identified specific proteins that are enriched in vesicles, and we have identified critical molecules that govern the internalization of vesicles into host cells. Using biochemical analysis of purified vesicles from cell-free culture supernatants, we have found that heat-labile enterotoxin, an important virulence factor of ETEC, is exported from the cells bound to the external surface of vesicles. Presented in this context, it is able to mediate the entry of the entire ETEC vesicle into human colorectal tissue culture cells. We have also discovered that the ability of vesicles to bind to specific cell types depends on their strain of origin: for example, P. aeruginosa vesicles produced by a strain that was cultured from the lungs of a patient with Cystic Fibrosis adhered better to lung than to gut epithelial cells, whereas a strain that was isolated from sera showed no such preference for lung cells. The vesicles stimulate epithelial cells and macrophages to elicit a cytokine response that is distinct from that of LPS (a major component of the vesicles) alone.
These studies will provide new insights into the membrane dynamics of gram-negative bacteria and consequently aid in the identification of new therapeutic targets for important human pathogens.
- Investigators in Pathogenesis of Infectious Diseases. Burroughs Wellcome Fund. 2002
- Triggers and Consequences of Bacterial Envelope Stress awarded by National Institutes of Health2012 - 2016
- Enterotoxin targeting and delivery mechanisms awarded by National Institutes of Health 2006 - 2012
- Heat-Labile Enterotoxin Secretion awarded by National Institutes of Health 2005 - 2008
- Detection of in vivo ETEC vesicle production awarded by National Institutes of Health 2003 - 2006
- Production and function of E Coli Vesicles awarded by National Institutes of Health 2002 - 2004
- Howard Hughes Postdoctoral Research Fellow, Biochemistry And Molecular Biolovy, University of California at Berkeley 1994 - 1997
- Postdoctoral Fellow, Molecular Microbiology, Washington University 1993
- Ph.D., Washington University 1993
- B.S., University of Washington 1986
- Bonnington, KE, and Kuehn, MJ. "Protein selection and export via outer membrane vesicles." Biochim Biophys Acta 1843, no. 8 (August 2014): 1612-1619. (Review) Full Text Link to Item
- Bonnington, KE, and Kuehn, MJ. "Protein selection and export via outer membrane vesicles." Biochimica et Biophysica Acta - Molecular Cell Research 1843, no. 8 (2014): 1612-1619. Full Text
- Schwechheimer, C, and Kuehn, MJ. "Synthetic effect between envelope stress and lack of outer membrane vesicle production in Escherichia coli." J Bacteriol 195, no. 18 (September 2013): 4161-4173. Full Text Link to Item
- Schwechheimer, C, Sullivan, CJ, and Kuehn, MJ. "Envelope control of outer membrane vesicle production in Gram-negative bacteria." Biochemistry 52, no. 18 (May 7, 2013): 3031-3040. Full Text Link to Item
- Chutkan, H, MacDonald, I, Manning, A, and Kuehn, MJ. "Quantitative and qualitative preparations of bacterial outer membrane vesicles." Methods in Molecular Biology 966 (2013): 259-272. Full Text