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Differential gene expression of the toll-like receptor-4 cascade and neutrophil function in early- and mid-lactating dairy cows

Mieke Stevens (UGent) , Luc Peelman (UGent) , Bart De Spiegeleer (UGent) , Adel Pezeshki (UGent) , Gerlinde Van de Walle (UGent) , Luc Duchateau (UGent) and Christian Burvenich (UGent)
(2011) JOURNAL OF DAIRY SCIENCE. 94(3). p.1277-1288
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Abstract
During early lactation, neutrophils display several reduced immune functions. Particularly, a delayed recruitment of neutrophils into the infected udder seems to be one of the underlying events involved in the severity of postpartum Escherichia coli intramammary infections. The purpose of this study was to analyze the effect of in vitro chemotaxis and diapedesis on the expression of toll-like receptor-4 (TLR4)-related genes in bovine blood neutrophils isolated from 10 early-lactating (EL) and 10 mid-lactating (ML) cows. Functional characterization of the neutrophil population was performed by measuring phagocytosis and production of reactive oxygen species (chemiluminescence). Messenger RNA was extracted from neutrophils, and the expression of TLR4 and associated genes in EL and ML cows was analyzed by reverse-transcription quantitative PCR. To study the effect of chemotaxis and diapedesis on the expression of genes of the TLR4 cascade, neutrophils were stimulated to (trans) migrate in response to C5a using in vitro models. Our salient findings were that both neutrophil migration in vitro and lactation stage induced significant changes in the expression of several genes of the TLR4 signaling cascade. Before migration, expression of TRAF6, ATF3, RELA, IL8, and C5aR were lower in EL than in ML cows. Diapedesis and chemotaxis induced an increase in expression of TLR4, ATF3, and IL8 in both EL and ML cows. Diapedesis resulted in a downregulation of Syk, a TLR4-associated gene, in ML cows. This study shows that the perturbations in neutrophil functions during EL are accompanied by modulation of TLR4 pathway genes. These data can contribute to the understanding of the mechanisms explaining the relationship between stage of lactation and risk of severe E. coli mastitis.
Keywords
BOVINE POLYMORPHONUCLEAR LEUKOCYTES, ESCHERICHIA-COLI MASTITIS, TYROSINE KINASE, PERIPARTURIENT PERIOD, BIOLOGICAL FUNCTIONS, SIGNAL-TRANSDUCTION, IMMUNE-RESPONSES, SYK, TOLL-LIKE-RECEPTOR-4, CHEMILUMINESCENCE, diapedesis, chemotaxis, toll-like receptor-4 (TLR4), neutrophil

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Chicago
Stevens, Mieke, Luc Peelman, Bart De Spiegeleer, Adel Pezeshki, Gerlinde Van de Walle, Luc Duchateau, and Christian Burvenich. 2011. “Differential Gene Expression of the Toll-like Receptor-4 Cascade and Neutrophil Function in Early- and Mid-lactating Dairy Cows.” Journal of Dairy Science 94 (3): 1277–1288.
APA
Stevens, Mieke, Peelman, L., De Spiegeleer, B., Pezeshki, A., Van de Walle, G., Duchateau, L., & Burvenich, C. (2011). Differential gene expression of the toll-like receptor-4 cascade and neutrophil function in early- and mid-lactating dairy cows. JOURNAL OF DAIRY SCIENCE, 94(3), 1277–1288.
Vancouver
1.
Stevens M, Peelman L, De Spiegeleer B, Pezeshki A, Van de Walle G, Duchateau L, et al. Differential gene expression of the toll-like receptor-4 cascade and neutrophil function in early- and mid-lactating dairy cows. JOURNAL OF DAIRY SCIENCE. 2011;94(3):1277–88.
MLA
Stevens, Mieke, Luc Peelman, Bart De Spiegeleer, et al. “Differential Gene Expression of the Toll-like Receptor-4 Cascade and Neutrophil Function in Early- and Mid-lactating Dairy Cows.” JOURNAL OF DAIRY SCIENCE 94.3 (2011): 1277–1288. Print.
@article{1235072,
  abstract     = {During early lactation, neutrophils display several reduced immune functions. Particularly, a delayed recruitment of neutrophils into the infected udder seems to be one of the underlying events involved in the severity of postpartum Escherichia coli intramammary infections. The purpose of this study was to analyze the effect of in vitro chemotaxis and diapedesis on the expression of toll-like receptor-4 (TLR4)-related genes in bovine blood neutrophils isolated from 10 early-lactating (EL) and 10 mid-lactating (ML) cows. Functional characterization of the neutrophil population was performed by measuring phagocytosis and production of reactive oxygen species (chemiluminescence). Messenger RNA was extracted from neutrophils, and the expression of TLR4 and associated genes in EL and ML cows was analyzed by reverse-transcription quantitative PCR. To study the effect of chemotaxis and diapedesis on the expression of genes of the TLR4 cascade, neutrophils were stimulated to (trans) migrate in response to C5a using in vitro models. Our salient findings were that both neutrophil migration in vitro and lactation stage induced significant changes in the expression of several genes of the TLR4 signaling cascade. Before migration, expression of TRAF6, ATF3, RELA, IL8, and C5aR were lower in EL than in ML cows. Diapedesis and chemotaxis induced an increase in expression of TLR4, ATF3, and IL8 in both EL and ML cows. Diapedesis resulted in a downregulation of Syk, a TLR4-associated gene, in ML cows. This study shows that the perturbations in neutrophil functions during EL are accompanied by modulation of TLR4 pathway genes. These data can contribute to the understanding of the mechanisms explaining the relationship between stage of lactation and risk of severe E. coli mastitis.},
  author       = {Stevens, Mieke and Peelman, Luc and De Spiegeleer, Bart and Pezeshki, Adel and Van de Walle, Gerlinde and Duchateau, Luc and Burvenich, Christian},
  issn         = {0022-0302},
  journal      = {JOURNAL OF DAIRY SCIENCE},
  keyword      = {BOVINE POLYMORPHONUCLEAR LEUKOCYTES,ESCHERICHIA-COLI MASTITIS,TYROSINE KINASE,PERIPARTURIENT PERIOD,BIOLOGICAL FUNCTIONS,SIGNAL-TRANSDUCTION,IMMUNE-RESPONSES,SYK,TOLL-LIKE-RECEPTOR-4,CHEMILUMINESCENCE,diapedesis,chemotaxis,toll-like receptor-4 (TLR4),neutrophil},
  language     = {eng},
  number       = {3},
  pages        = {1277--1288},
  title        = {Differential gene expression of the toll-like receptor-4 cascade and neutrophil function in early- and mid-lactating dairy cows},
  url          = {http://dx.doi.org/10.3168/jds.2010-3563},
  volume       = {94},
  year         = {2011},
}

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