Novel Dietary Proteins Selectively Affect Intestinal Health In Vitro after Clostridium difficile-Secreted Toxin A Exposure.

Paulus G M Jochems,Johan Garssen,Jeroen Van Bergenhenegouwen,Harry J Wichers,Pascale C S Rietveld,Coen Govers,Monic M M Tomassen,Rosalinde Masereeuw

doi:10.3390/nu12092782

Abstract

Bacterial gastroenteritis forms a burden on a global scale, both socially and economically. The Gram-positive bacterium Clostridium difficile is an inducer of gastrointestinal bacterial infections, often triggered following disruption of the microbiota by broad-spectrum antibiotics to treat other conditions. The clinical manifestatiaons, e.g., diarrhea, are driven by its toxins secretion, toxin A (TcdA) and toxin B (TcdB). Current therapies are focused on discontinuing patient medication, including antibiotics. However, relapse rates upon therapy are high (20–25%). Here, eighteen dietary proteins were evaluated for their capacity to restore gut health upon C. difficile-derived TcdA exposure. We used bioengineered intestinal tubules to assess proteins for their beneficial effects by examining the epithelial barrier, cell viability, brush-border enzyme activity, IL-6 secretion, IL-8 secretion and nitric oxide (NO) levels upon TcdA challenge. TcdA effectively disrupted the epithelial barrier, increased mitochondrial activity, but did not affect alkaline phosphatase activity, IL-6, IL-8 and NO levels. Intervention with dietary proteins did not show a protective effect on epithelial barrier integrity or mitochondrial activity. However, bovine plasma and potato protein increased alkaline phosphatase activity, egg-white protein increased IL-6 and IL-8 release and wheat, lesser mealworm and yeast protein increased NO levels after TcdA exposure. Hence, dietary proteins can influence parameters involved in intestinal physiology and immune activation suggesting that supplementation with specific dietary proteins may be of benefit during C. difficile infections.

Highlights

Bacterial gastroenteritis is a common pathology worldwide, is associated with 1.5–2.5 million deaths per year and forms a great burden for society on a social and economic scale [1]
None of the potential proteins could prevent TcdA-induced barrier impairment (Figure S1 (Supplementary Materials)). To investigate their impact on intestinal epithelial health in more detail, these eighteen dietary protein sources were subdivided into animal, plantor alternative-based sources and evaluated for their recovery effect after TcdA-induced disruption
Experiment-ready bioengineered intestinal tubules were viable and demonstrated to have a functional epithelial barrier including a clear zonula occludens-1 (ZO-1) network and increased alkaline phosphatase activity compared to the negative control (Figure 1A,C–E)

Summary

Introduction

Bacterial gastroenteritis is a common pathology worldwide, is associated with 1.5–2.5 million deaths per year and forms a great burden for society on a social and economic scale [1]. Clinical manifestations of gastrointestinal (GI) bacterial infections are vomiting, abdominal pain, nausea and diarrhea [1]. This can be induced by various mechanism such as mucosal invasion by pathogens and bacterial toxin production [1]. C. Difficile infections are a healthcare-associated problem and estimated to increase hospitalization costs by approximately $1.5 billion in the United States alone [3]. Many of the clinical manifestations are the result of the C. difficile release of toxin A and toxin

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