An experimental platform using human intestinal epithelial cell lines to differentiate between hazardous and non-hazardous proteins
| Autor: | Marc S. Harper, Raymond Layton, Waheed Pirzai, Bryan P. Hurley, Cindi Zimmermann, Bryan Delaney, Alex D. Eaton, Jason M. Roper, Gregory S. Ladics |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Bacterial Toxins Biology medicine.disease_cause Toxicology 03 medical and health sciences 0404 agricultural biotechnology Listeria monocytogenes Lectins Enterotoxigenic Escherichia coli medicine Animals Humans Neoplasms Glandular and Epithelial Cytotoxicity Cells Cultured Cell Proliferation Venoms Toxin Albumin Listeriolysin O 04 agricultural and veterinary sciences General Medicine Allergens 040401 food science Intestines 030104 developmental biology Biochemistry Mastoparan Cytokines Cattle Streptolysin Dietary Proteins Food Science |
| Zdroj: | Food and Chemical Toxicology. 92:75-87 |
| ISSN: | 0278-6915 |
| DOI: | 10.1016/j.fct.2016.04.003 |
| Popis: | Human intestinal epithelial cell lines (T84, Caco-2, and HCT-8) grown on permeable Transwell™ filters serve as models of the gastrointestinal barrier. In this study, this in vitro model system was evaluated for effectiveness at distinguishing between hazardous and non-hazardous proteins. Indicators of cytotoxicity (LDH release, MTT conversion), monolayer barrier integrity ([(3)H]-inulin flux, horseradish peroxidase flux, trans-epithelial electrical resistance [TEER]), and inflammation (IL-8, IL-6 release) were monitored following exposure to hazardous or non-hazardous proteins. The hazardous proteins examined include streptolysin O (from Streptococcus pyogenes), Clostridium difficile Toxins A and B, heat-labile toxin from enterotoxigenic Escherichia coli, listeriolysin O (from Listeria monocytogenes), melittin (from bee venom), and mastoparan (from wasp venom). Non-hazardous proteins included bovine and porcine serum albumin, bovine fibronectin, and ribulose bisphosphate carboxylase/oxygenase (RuBisco) from spinach. Food allergenic proteins bovine milk β-lactoglobulin and peanut Ara h 2 were also tested as was the anti-nutritive food protein wheat germ agglutinin. Results demonstrated that this model system effectively distinguished between hazardous and non-hazardous proteins through combined analysis of multiple cells lines and assays. This experimental strategy may represent a useful adjunct to multi-component analysis of proteins with unknown hazard profiles. |
| Databáze: | OpenAIRE |
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