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Cytokine response of bovine mammary gland epithelial cells to Escherichia coli, coliform culture filtrate, or lipopolysaccharide

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  • 1 Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI 53706.
  • | 2 Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI 53706.
  • | 3 Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI 53706.

Abstract

Objective—To define the cytokine response of a cultured mammary gland epithelial cell line (ie, Mac-T) when incubated with Escherichia coli or its products.

Sample Population—Mac-T cells and E coli from cows with mastitis.

Procedure—Mac-T cells were incubated with E coli or its products. The cytokine response of Mac-T cells to these treatments was quantified by measuring mRNA content of interleukin (IL)-1α, IL-1β, IL-8, and tumor necrosis factor (TNF)-α by use of a quantitative reverse transcriptase-polymerase chain reaction assay. The amount of TNF-α secreted was also measured.

Results—Treatment with E coli or its products resulted in significant increases in IL-1α, IL-8, and TNF-α mRNA content in Mac-T cells. This increase was reversible when culture filtrate was incubated with polymyxin B. The amount of IL-1β mRNA in Mac-T cells increased only slightly over baseline after treatment with E coli or its products, but this increase was not diminished by incubation of E coli filtrate with polymyxin B.

Conclusions and Clinical Relevance—Incubation of Mac-T cells with E coli or its products resulted in increased amounts of IL-1α, IL-8, and TNF-α mRNA. Inhibition of this response by incubation of culture filtrate with polymyxin B suggested that lipopolysaccharide was the main bacterial product that stimulated the cytokine response. The small increase in IL-1β content in Mac-T cells incubated with E coli or its products suggested that this cytokine had a smaller role in the Mac-T cell response to E coli. (Am J Vet Res 2005;66:1590–1597)

Abstract

Objective—To define the cytokine response of a cultured mammary gland epithelial cell line (ie, Mac-T) when incubated with Escherichia coli or its products.

Sample Population—Mac-T cells and E coli from cows with mastitis.

Procedure—Mac-T cells were incubated with E coli or its products. The cytokine response of Mac-T cells to these treatments was quantified by measuring mRNA content of interleukin (IL)-1α, IL-1β, IL-8, and tumor necrosis factor (TNF)-α by use of a quantitative reverse transcriptase-polymerase chain reaction assay. The amount of TNF-α secreted was also measured.

Results—Treatment with E coli or its products resulted in significant increases in IL-1α, IL-8, and TNF-α mRNA content in Mac-T cells. This increase was reversible when culture filtrate was incubated with polymyxin B. The amount of IL-1β mRNA in Mac-T cells increased only slightly over baseline after treatment with E coli or its products, but this increase was not diminished by incubation of E coli filtrate with polymyxin B.

Conclusions and Clinical Relevance—Incubation of Mac-T cells with E coli or its products resulted in increased amounts of IL-1α, IL-8, and TNF-α mRNA. Inhibition of this response by incubation of culture filtrate with polymyxin B suggested that lipopolysaccharide was the main bacterial product that stimulated the cytokine response. The small increase in IL-1β content in Mac-T cells incubated with E coli or its products suggested that this cytokine had a smaller role in the Mac-T cell response to E coli. (Am J Vet Res 2005;66:1590–1597)