Bovine lactoferrin digested with human gastrointestinal enzymes inhibits replication of human echovirus 5 in cell culture |
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Authors: | Camilla B. Furlund Anja B. Kristoffersen Tove G. Devold Gerd E. Vegarud Christine M. Jonassen |
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Affiliation: | 1. Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, 1432 Aas, Norway;2. Department of Medical Microbiology, Akershus University Hospital, 1478 Loerenskog, Norway;3. Department of Laboratory Services, The Norwegian Veterinary Institute, 0106 Oslo, Norway;4. Department of Health Surveillance, The Norwegian Veterinary Institute, 0106 Oslo, Norway |
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Abstract: | Many infant formulas are enriched with lactoferrin (Lf) because of its claimed beneficial effects on health. Native bovine Lf (bLf) is known to inhibit in vitro replication of human enteroviruses, a group of pathogenic viruses that replicate in the gut as their primary infection site. On the basis of a model digestion and human gastrointestinal enzymes, we hypothesized that bLf could retain its antiviral properties against enterovirus in the gastrointestinal tract, either as an intact protein or through bioactive peptide fragments released by digestive enzymes. To test our hypothesis, bLf was digested with human gastric juice and duodenal juice in a 2-step in vitro digestion model. Two gastric pH levels and reduction conditions were used to simulate physiological conditions in adults and infants. The antiviral activity of native bLf and of the digested fractions was studied on echovirus 5 in vitro, using various assay conditions, addressing several mechanisms for replication inhibition. Both native and digested bLf fractions revealed a significant inhibitory effect, when added before or simultaneously with the virus onto the cells. Furthermore, a significant stronger sustained antiviral effect was observed when bLf was fully digested in the gastric phase with fast pH reduction to 2.5, compared with native bLf, suggesting the release of antiviral peptides from bLf during the human digestion process. In conclusion, this study demonstrates that bLf may have a role in the prevention of human gastrointestinal virus infection under physiological conditions and that food containing bLf may protect against infection in vivo. |
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Keywords: | bLf, bovine lactoferrin CPE, cytopathic effect Ct, cycle threshold DPBS, Dulbecco phosphate-buffered saline F, fast reduction in gastric pH HCMV, human cytomegalovirus HDJ, human duodenal juice HGJ, human gastric juice HPV, human papillomavirus HS, heparan sulfate HSV, herpes simplex virus Lf, lactoferrin Lfcin, lactoferricin LfR, lactoferrin receptors LMW, low molecular weight MEM, Eagle minimal essential medium MOI, multiplicity of infection S, slow reduction in gastric pH SDS-PAGE, sodium dodecyl sulfate&ndash polyacrylamide gel electrophoresis Si-LfR, small intestinal lactoferrin receptor |
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