Gastroenterology

Gastroenterology

Volume 126, Issue 2, February 2004, Pages 488-498
Gastroenterology

Basic-alimentary tract
High-molecular-weight polyethylene glycol prevents lethal sepsis due to intestinal Pseudomonas aeruginosa

https://doi.org/10.1053/j.gastro.2003.11.011Get rights and content

Abstract

Background & Aims: During stress, erosion of protective intestinal mucus occurs in association with adherence to and disruption of the intestinal epithelial barrier by invading opportunistic microbial pathogens. The aims of this study were to test the ability of a high-molecular-weight polyethylene glycol compound, polyethylene glycol 15–20, to protect the intestinal epithelium against microbial invasion during stress. Methods: The ability of polyethylene glycol 15–20 to protect the intestinal epithelium against the opportunistic pathogen Pseudomonas aeruginosa was tested in cultured Caco-2 cells. Bacterial virulence gene expression, bacterial adherence, and transepithelial electrical resistance were examined in response to apical inoculation of P. aeruginosa onto Caco-2 cells. Complementary in vivo studies were performed in a murine model of lethal sepsis due to intestinal P. aeruginosa in which surgical stress (30% hepatectomy) was combined with direct inoculation of P. aeruginosa into the cecum. Results: High-molecular-weight polyethylene glycol (polyethylene glycol 15–20) conferred complete protection against the barrier-dysregulating effects of P. aeruginosa in Caco-2 cells. Intestinal application of polyethylene glycol 15–20 in stressed mice protected against the lethal effects of intestinal P. aeruginosa. Mechanisms of this effect seem to involve the ability of polyethylene glycol 15–20 to distance P. aeruginosa from the intestinal epithelium and render it completely insensate to key environmental stimuli that activate its virulence. Conclusions: High-molecular-weight polyethylene glycol has the potential to function as a surrogate mucin within the intestinal tract of a stressed host by inhibiting key interactive events between colonizing microbes and their epithelial cell targets.

Section snippets

Bacterial strains, epithelial cell lines, and polyethylene glycol solutions

P. aeruginosa strain 27853 (PA27853; American Type Culture Collection, Manassas, VA) is a nonmucoid clinical strain originally isolated from a blood culture. All experiments were performed with Caco-2/C2bbe, or C2, cells (passage 51–68) and were obtained as a generous gift of Dr. Mark Mooseker (Yale University, New Haven, CT). C2 cells are Caco-2 subclones that are highly differentiated intestinal epithelial cells.10 Two PEGs of different molecular weights were purchased from Sigma (St. Louis,

Results

PEG 15–20 protects against lethal gut-derived sepsis due to P. aeruginosa in mice after 30% hepatectomy. Direct cecal injection of PA27853 into mice who underwent a 30% surgical hepatectomy resulted in a state of clinical sepsis with no survivors at 48 hours (Figure 2A). Mice undergoing sham laparotomy without hepatectomy (controls), similarly injected with P. aeruginosa, survived completely without any clinical signs of sepsis (Figure 2A). To determine the ability of PEG solutions to prevent

Discussion

In this study, the ability of PEG 15–20 to protect the intestinal epithelium against an opportunistic colonizing pathogen was assessed. Specifically, the ability of PEG to inactivate the lethal effects of intestinal P. aeruginosa, one of the more common causes of fatal gut-derived sepsis in an immunocompromised host, was tested.19, 20, 21 Data from this show that there are striking effects of PEG 15–20 that parallel the function of intestinal mucus. Because intestinal mucus exerts its effects

Acknowledgements

The authors thank Dr. Klaus Winzer and Dr. Paul Williams, University of Nottingham, for helpful technical advice; Raphael Lee, M.D., Sc.D., director of the Center for Molecular Repair, University of Chicago, and William Cromie, M.D., for helpful conversations; and Shirley Bond, Cancer Center Digital Light Microscopy Facility at the University of Chicago, for technical assistance. The Escherichia coli/Pseudomonas aeruginosa shuttle vector pUCP24 was a gift of Dr. H. P. Schweizer, Colorado State

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    Supported by National Institutes of Health Grants RO1 GM62344-01 (to J.C.A.) and DK619131 (to J.R.T.), Digestive Diseases Research Core Center Grant DK 47722 (to E.B.C.), the David and Lucile Pachard Foundation Grant 99-1465 (to K.Y.C.L.), and the University of Chicago Materials Research Science and Engineering Centers Program of the National Science Foundation under Award DMR 9808595 (to K.Y.C.L.).

    1

    L.W. and O.Z. contributed equally to this work.

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