Lipopolysaccharide and lipoteichoic acid induce different innate immune responses in bovine mammary epithelial cells

Strandberg, Ylva; Gray, Christian; Vuocolo, Tony; Donaldson, Laurelea; Broadway, Mary M.; Tellam, Ross L.

doi:10.1016/j.cyto.2005.02.010

Cited by 249 publications

(273 citation statements)

References 59 publications

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“…It seems that it is not necessary to strongly upregulate the TLR pathway components for an efficient immune response. Strandberg et al (2005) found a similar weak TLR activation in bovine MECs upon LPS stimulation and still came to the conclusion that a functioning and locally effective immune system was present.…”

Section: Pathogen Differencesmentioning

confidence: 85%

“…It could have been too low, taking into account that there is a dose-dependent immune response of pbMEC to lipopolysaccharide (LPS) and S. aureus (Wellnitz and Kerr, 2004) and a study by Swanson et al (2009) with pbMEC from tissue showed an upregulation in four of nine measured immune genes to S. aureus with a much higher MOI of 1000. Another possibility is that we missed the proper time frame of the immune response: one study showed an early immune response of MECs to S. aureus that decreased to resting levels after 8 to 16 h (Strandberg et al, 2005). Our bacteria had been isolated from a clinical case of mastitis and were shown to have elicited weak but measurable symptoms of mastitis when administered in vivo intra-mammary (Petzl et al, 2008); thus, the question remains whether this strain exhibits sufficient virulence only in a live, but not in a heat-inactivated form.…”

Section: Pathogen Differencesmentioning

confidence: 99%

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Microfluidic high-throughput RT-qPCR measurements of the immune response of primary bovine mammary epithelial cells cultured from milk to mastitis pathogens

Sorg

Danowski

Korenkova

et al. 2013

Animal

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Bovine mastitis, the inflammation of the udder, is a major problem for the dairy industry and for the welfare of the animals. To better understand this disease, and to implement two special techniques for studying mammary gland immunity in vitro, we measured the innate immune response of primary bovine mammary epithelial cells (pbMEC) from six Brown Swiss cows after stimulation with the heat-inactivated mastitis pathogens, Escherichia coli 1303 and Staphylococcus aureus 1027. The cells were extracted and cultivated from milk instead of udder tissue, which is usually done. The advantages of this technique are non-invasiveness and less contamination by fibroblasts. For the first time, pbMEC gene expression (GE) was measured with a microfluidic high-throughput real-time reverse transcription-quantitative PCR platform, the BioMark HD TM system from Fluidigm. In addition to the physiological analysis, the precision and suitability of this method was evaluated in a large data set. The mean coefficient of variance (6 s.e.) between repeated chips was 4.3 6 0.4% for highly expressed and 3.3 6 0.4% for lowly expressed genes. Quantitative PCR (qPCR) replicate deviations were smaller than the cell culture replicate deviations, indicating that biological and cell culture differences could be distinguished from the background noise. Twenty-two genes (complement system, chemokines, inflammatory cytokines, antimicrobial peptides, acute phase response and toll-like receptor signalling) were differentially expressed (P , 0.05) with E. coli. The most upregulated gene was the acute phase protein serum amyloid A3 with 618-time fold. S. aureus slightly induced CCL5, IL10, TLR4 and S100A12 expression and failed to elicit a distinct overall innate immune response. We showed that, with this milk-derived pbMEC culture and the high-throughput qPCR technique, it is possible to obtain similar results in pbMEC expression as with conventional PCR and with satisfactory precision so that it can be applied in future GE studies in pbMEC.Keywords: bovine mastitis, gene expression profiling, microfluidic qPCR, primary bovine mammary epithelial cells, innate immune response ImplicationsWe show that a time-and cost-efficient high-throughput quantitative PCR (qPCR) system, applied on primary bovine mammary epithelial cells (pbMEC) cultured from milk, is a convenient alternative to the two major standard procedures in measuring gene expression. We obtained similar results as studies with pbMEC from udder tissue and measurements on DNA microarrays or conventional qPCR. We suggest that the milk-derived pbMEC culture and the microfluidic high-throughput qPCR system could be applied in future experiments with pbMEC.

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Section: Pathogen Differencesmentioning

confidence: 85%

Section: Pathogen Differencesmentioning

confidence: 99%

Microfluidic high-throughput RT-qPCR measurements of the immune response of primary bovine mammary epithelial cells cultured from milk to mastitis pathogens

Sorg

Danowski

Korenkova

et al. 2013

Animal

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“…It has been shown that cultures of bovine MEC respond to S. aureus LTA [51]. The receptor complex involved in the recognition of LTA by MEC has not been characterized.…”

Section: Discussionmentioning

confidence: 99%

“…The recognition of LTA by leucocytes requires TLR2 and is improved by interaction with CD14 [45]. Bovine mammary tissue and bovine MEC express transcripts of the gene coding TLR2 [16,51], and the protein is expressed on bovine MEC [24]. Bovine MEC express transcripts for CD14 [51], and the protein is found on the milk fat globule membrane [42], which derives from the apical face of MEC.…”

Section: Discussionmentioning

confidence: 99%

“…Bovine mammary tissue and bovine MEC express transcripts of the gene coding TLR2 [16,51], and the protein is expressed on bovine MEC [24]. Bovine MEC express transcripts for CD14 [51], and the protein is found on the milk fat globule membrane [42], which derives from the apical face of MEC. The scavenger receptor CD36 has been reported to facilitate the response of monocytes and T lymphocytes to LTA [23,28].…”

Section: Discussionmentioning

confidence: 99%

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Staphylococcus aureuslipoteichoic acid triggers inflammation in the lactating bovine mammary gland

et al. 2008

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-The response of the bovine mammary gland to lipoteichoic acid (LTA), which is a major pathogen-associated molecular pattern of Gram-positive bacteria, was investigated by infusing purified Staphylococcus aureus LTA in the lumen of the gland. LTA was able to induce clinical mastitis at the dose of 100 g/quarter, and a subclinical inflammatory response at 10 g/quarter. The induced inflammation was characterized by a prompt and massive influx of neutrophils in milk. LTA proved to induce strongly the secretion of the chemokines CXCL1, CXCL2, CXCL3 and CXCL8, which target mainly neutrophils. The complement-derived chemoattractant C5a was generated in milk only with the highest dose of LTA (100 g). The pro-inflammatory cytokine IL-1 was induced in milk, but there was very little if any TNF-and no IFN-. The re-assessment of CXCL8 concentrations in milk whey of quarters previously challenged with S. aureus, by using an ELISA designed for bovine CXCL8, showed that this chemokine was induced in milk, contradicting previous reports. Overall, S. aureus LTA elicited mammary inflammatory responses that shared several attributes with S. aureus mastitis. Purified LTA looks promising as a convenient tool to investigate the inflammatory and immune responses of the mammary gland to S. aureus. Staphylococcus aureus / mastitis / cattle / lipoteichoic acid / chemokine

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Inhibition of histone deacetylases is the major pathway mediated by astaxanthin to antagonize LPS‐induced inflammatory responses in mammary epithelial cells

Venkidasamy

Thiruvengadam

Thirupathi

et al. 2020

J Biochem & Molecular Tox

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Mastitis is a major inflammatory response of the mammary gland due to various pathogenic invasions and is a serious disease that affects the production yield and health status of cows. Astaxanthin (AST), a xanthophyll carotenoid, is a secondary metabolite synthesized by microalgae and yeasts that has been reported to suppress various inflammatory responses. However, the protective effect of AST on lipopolysaccharide (LPS)‐induced mammary epithelial cells has not yet been reported. The present study results indicated that AST treatment markedly attenuated the oxidative stress markers and nitric oxide (NO) while improving the anti‐oxidant enzymes in LPS exposed cells. On the other hand, LPS‐exposed cells showed nuclear translocation of nuclear factor‐κB (NF‐κB) with the activation of inflammatory cytokines such as monocyte chemoattractant protein‐1, tumor necrosis factor‐α, interferon‐γ, and interleukin‐6 (IL‐6). In addition, mRNA expression analysis revealed that the histone deacetylase (HDAC) ‐1, ‐2, ‐3, ‐6, ‐7 and pentraxin 3 (PTX3) expressions were increased in the LPS group. Furthermore, the activity of HDAC was increased to 2‐fold with a significant reduction in the histone acetyltransferase activity in cells exposed to LPS. However, AST was able to inhibit the nuclear translocation of NF‐κB with attenuated HDAC activity. Intriguingly, HDAC inhibition studies demonstrated that the cytokines such as IL‐4, IL‐8, granulocyte‐mcrophage colony stimulating factor, C‐reactive protein, IL‐17A, and IL‐22 were significantly suppressed which were upregulated in LPS treatment; while AST was found acting by improving the anti‐inflammatory cytokine IL‐10, and thioredoxin reductase levels. Collectively, these findings provide novel insights into the role of HDACs in regulating cellular processes involved in the pathogenesis of LPS‐induced mastitis as well as the potential use of AST as a therapeutic in treatment for controlling disease progression.

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Lipopolysaccharide and lipoteichoic acid induce different innate immune responses in bovine mammary epithelial cells

Cited by 249 publications

References 59 publications

Microfluidic high-throughput RT-qPCR measurements of the immune response of primary bovine mammary epithelial cells cultured from milk to mastitis pathogens

Microfluidic high-throughput RT-qPCR measurements of the immune response of primary bovine mammary epithelial cells cultured from milk to mastitis pathogens

Staphylococcus aureuslipoteichoic acid triggers inflammation in the lactating bovine mammary gland

Inhibition of histone deacetylases is the major pathway mediated by astaxanthin to antagonize LPS‐induced inflammatory responses in mammary epithelial cells

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