Interleukin 8 response by bovine mammary epithelial cells to lipopolysaccharide stimulation
To determine whether an established bovine mammary epithelial cell line expresses interleukin 8 (IL-8) mRNA and synthesizes antigenic IL-8 in response to lipopolysaccharide (LPS) stimulation. A bovine mammary epithelial cell line (MAC-T). mRNA was isolated from cells stimulated with graded concentra...
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Published in | American journal of veterinary research Vol. 59; no. 12; p. 1563 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
United States
01.12.1998
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Subjects | |
Online Access | Get more information |
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Summary: | To determine whether an established bovine mammary epithelial cell line expresses interleukin 8 (IL-8) mRNA and synthesizes antigenic IL-8 in response to lipopolysaccharide (LPS) stimulation.
A bovine mammary epithelial cell line (MAC-T).
mRNA was isolated from cells stimulated with graded concentrations of LPS. The first strand of IL-8 cDNA was synthesized, using a reverse transcriptase (RT) reaction with a specific oligonucleotide. Amplification of IL-8 cDNA was obtained by use of polymerase chain reaction (PCR). The MAC-T-derived antigenic IL-8 was quantified by use of a commercial anti-human IL-8 kit in a sandwich ELISA.
RT-PCR revealed expression of MAC-T-derived mRNA within the first hour after stimulation with LPS. Expression of IL-8 mRNA was correlated to production of IL-8 protein detected in medium by use of the sandwich ELISA. Amounts of antigenic IL-8 increased in a dose- and time-dependent manner, and were maximal (57 pg/ml) at 48 hours after stimulation with 20 microg of LPS/ml.
MAC-T cells secrete IL-8 in response to stimulation with LPS in a dose- and time- dependent manner. The results were consistent with our hypothesis that mammary gland epithelial cells can be a source of IL-8 during the early stage of mastitis. Therefore, IL-8 may have a pivotal role in resolving bacterial infections. |
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Bibliography: | L70 L10 1999009536 |
ISSN: | 0002-9645 1943-5681 |
DOI: | 10.2460/ajvr.1998.59.12.1563 |