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Reversal of Pathogen-Induced Barrier Defects in Intestinal Epithelial Cells by Contra-pathogenicity Agents

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Abstract

Background

Environmental enteropathy (EE) is associated with stunting, impairment of responses to oral vaccines, and other adverse health consequences in young children throughout the developing world. EE is characterized by chronic low-grade intestinal inflammation and disrupted epithelial barrier integrity, partly resulting from dysregulation of tight junction proteins, observed in other enteropathies such as celiac disease. During EE, this dysregulation of tight junction expression amplifies translocation of pathogenic bacteria across the intestinal mucosa.

Aims

The aim was to determine whether enteropathogen-mediated epithelial barrier failure can be ameliorated using contra-pathogenicity therapies.

Methods

Intestinal epithelial barrier damage was assessed in Caco-2 cells incubated with three important enteropathogens identified in EE patients: Enteropathogenic Escherichia coli (EPEC), Citrobacter rodentium (C. rodentium), and Cryptosporidium parvum (C. parvum). Potential therapeutic molecules were tested to detect effects on transepithelial resistance (TER), bacterial translocation (BT), claudin-4 expression, and regulation of the inflammatory cytokine response.

Results

All three enteropathogens compared to uninfected cells, reduced TER (EPEC; p < 0.0001, C. rodentium; p < 0.0001, C. parvum; p < 0.0007), reduced claudin-4 expression, and permitted BT (EPEC; p < 0.0001, C. rodentium; p < 0.0001, C. parvum; p < 0.0003) through the monolayer. Zinc, colostrum, epidermal growth factor, trefoil factor 3, resistin-like molecule-β, hydrocortisone, and the myosin light chain kinase inhibitor ML7 (Hexahydro-1-[(5-iodo-1-naphthalenyl)sulfonyl]-1H-1,4-diazepine hydrochloride); ML7) improved TER (up to 70%) and decreased BT (as much as 96%). Only zinc demonstrated modest antimicrobial activity.

Conclusion

The enteropathogens impaired intestinal–epithelial barrier integrity with dysregulation of claudin-4 and increased bacterial translocation. Enteropathogen-mediated damage was reduced using contra-pathogenicity agents which mitigated the effects of pathogens without direct antimicrobial activity.

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Acknowledgments

We gratefully acknowledge funding from Barts and The London Charity, and David Wareham (Blizard Institute, Barts and The London School of Medicine, UK) for a gift of E. coli K12.

Funding

Funding for this study was from Bart’s Charity and provided support in the form of salaries for authors [NC] but did not have any additional role in the study design, data collection, and analysis, decision to publish, or preparation of the manuscript. ME is an employee of Takeda Pharmaceuticals—This funder provided support in the form of salaries for authors [ME] but did not have any additional role in the study design, data collection, and analysis, decision to publish, or preparation of the manuscript. GJS is currently in receipt of funding from Takeda Pharmaceuticals, BBSRC together with 31, Benevolent and the Dunhill Foundation. He acts as a scientific advisor to 3 Takeda Pharmaceuticals and Zealand Pharma. “These funders provided support in the form of salaries for authors [GJS] but did not have any additional role in the study design, data collection, and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. Barts and The London School of Medicine, Queen Mary, University of London, London, E1 2AT, UK

    Naheed Choudhry, Flora Scott, Gareth J. Sanger & Paul Kelly

  2. Gastroenterology Drug Discovery Unit, Takeda California, Inc., 10410 Science Center Drive, San Diego, CA, 92121, USA

    Meghan Edgar & Gareth J. Sanger

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  1. Naheed Choudhry
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  2. Flora Scott
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  3. Meghan Edgar
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Correspondence to Naheed Choudhry.

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Choudhry, N., Scott, F., Edgar, M. et al. Reversal of Pathogen-Induced Barrier Defects in Intestinal Epithelial Cells by Contra-pathogenicity Agents. Dig Dis Sci 66, 88–104 (2021). https://doi.org/10.1007/s10620-020-06121-9

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