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

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)-1alpha, IL-1beta, IL-8, and tumor necrosis factor (TNF)-alpha by use of a quantitative reverse transcriptase-polymerase chain reaction assay. The amount of TNF-alpha secreted was also measured. RESULTS: Treatment with E coli or its products resulted in significant increases in IL-1alpha, IL-8, and TNF-alpha mRNA content in Mac-T cells. This increase was reversible when culture filtrate was incubated with polymyxin B. The amount of IL-1beta 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 IL1alpha, IL-8, and TNF-alpha 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-1beta 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.