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

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.