In vitro effects of lactoferrin on lipopolysaccharide-induced proliferation, gene expression, and prostanoid production by bovine peripheral blood mononuclear cells

Maisie E. Dawes Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri, Columbia, MO 65211.

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Jeff W. Tyler Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, University of Missouri, Columbia, MO 65211.

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Antoinette E. Marsh Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri, Columbia, MO 65211.

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Robert L. Larson Department of Veterinary Extension, College of Veterinary Medicine, University of Missouri, Columbia, MO 65211.

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Barry J. Steevens Department of Animal Sciences, College of Agriculture, Food and Natural Resources, University of Missouri, Columbia, MO 65211.

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Jeffrey Lakritz Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, University of Missouri, Columbia, MO 65211.

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Abstract

Objective—To evaluate the effect of lactoferrin on lipopolysaccharide (LPS)-induced proliferation of bovine peripheral blood mononuclear cells (PBMCs), gene expression of inflammatory mediators, and production of prostanoids in vitro.

Sample Population—PBMCs isolated from 15 Holstein bull calves.

Procedures—Mixed populations of PBMCs were isolated by differential centrifugation. Proliferation assays were conducted in 96-well plates designed to allow addition of lactoferrin (200 ng/mL) with and without LPS (1 μg/mL) in a checkerboard design. Incorporation of 3H-thymidine was used to determine proliferation of PBMCs. Prostaglandin E2 production was determined in culture-conditioned medium by use of enzyme immunoassay. Effects of lactoferrin on LPS-induced gene expression of cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 were monitored by use of PCR assays.

Results—Lactoferrin supplementation significantly reduced LPS-induced incorporation of 3H-thymidine and production of prostaglandin E2 by PBMCs. Lactoferrin reduced LPS-induced expression of COX-2 and MMP-9 mRNA.

Conclusions and Clinical Relevance—Lactoferrin reduced LPS-induced cellular proliferation, inflammatory mediator gene expression, and prostaglandin E2 production by bovine PBMCs in vitro. These effects may be beneficial in reducing the impact of endotoxemia in neonates.

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