Differential responsiveness of Holstein and Angus dermal fibroblasts to LPS challenge occurs without major differences in the methylome

Benjamin, Aimee; Green, Benjamin B.; Crooker, B.A.; McKay, Stephanie; Kerr, David E.

doi:10.1186/s12864-016-2565-x

Cited by 14 publications

(16 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Just as with the age difference, we have shown that dermal fibroblasts collected from the Holstein breed are much more responsive to LPS than fibroblasts collected from the Angus breed [26]. To determine whether similar gene expression differences could be detected between breeds, RT-qPCR was performed on age-matched dermal fibroblast cultures at 0, 2 and 8 h post-LPS treatment.…”

Section: Resultsmentioning

confidence: 99%

Genome-wide methylation analysis reveals differentially methylated loci that are associated with an age-dependent increase in bovine fibroblast response to LPS

Korkmaz

Kerr

2017

BMC Genomics

Self Cite

View full text Add to dashboard Cite

BackgroundDifferences in DNA methylation are known to contribute to the development of immune-related disorders in humans but relatively little is known about how methylation regulates immune function in cattle. Utilizing whole-transcriptome analyses of bovine dermal fibroblasts, we have previously identified an age and breed-dependent up-regulation of genes within the toll-like receptor 4 (TLR4) pathway that correlates with enhanced fibroblast production of IL-8 in response to lipopolysaccharide (LPS). Age-dependent differences in IL-8 production are abolished by treatment with 5-aza-2-deoxycytidine and Trichostatin A (AZA-TSA), suggesting epigenetic regulation of the innate response to LPS. In the current study, we performed reduced representation bisulfite sequencing (RRBS) on fibroblast cultures isolated from the same animals at 5- and 16-months of age to identify genes that exhibit variable methylation with age. To validate the role of methylation in gene expression, six innate response genes that were hyper-methylated in young animals were assessed by RT-qPCR in fibroblasts from animals at different ages and from different breeds.ResultsWe identified 14,094 differentially methylated CpGs (DMCs) that differed between fibroblast cultures at 5- versus 16-months of age. Of the 5065 DMCs that fell within gene regions, 1117 were located within promoters, 1057 were within gene exons and 2891 were within gene introns and 67% were more methylated in young cultures. Transcription factor enrichment of the promoter regions hyper-methylated in young cultures revealed significant regulation by the key pro-inflammatory regulator, NF-κB. Additionally, five out of six chosen genes (PIK3R1, FES, NFATC1, TNFSF13 and RORA) that were more methylated in young cultures showed a significant reduction in expression post-LPS treatment in comparison with older cultures. Two of these genes, FES and NFATC1, were similarly down-regulated in Angus cultures that also exhibit a low LPS response phenotype.ConclusionsOur study has identified immune-related loci regulated by DNA methylation in cattle that may contribute to differential cellular response to LPS, two of which exhibit an identical expression profile in both low-responding age and breed phenotypes. Methylation biomarkers of differential immunity may prove useful in developing selection strategies for replacement cows that are less susceptible to severe infections, such as coliform mastitis.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-017-3796-1) contains supplementary material, which is available to authorized users.

show abstract

Section: Resultsmentioning

confidence: 99%

Genome-wide methylation analysis reveals differentially methylated loci that are associated with an age-dependent increase in bovine fibroblast response to LPS

Korkmaz

Kerr

2017

BMC Genomics

Self Cite

View full text Add to dashboard Cite

show abstract

“…Although expression of SAA3 was higher in HR animals at all 3 time points, no significant differences were measured, and the weak induction of SAA3 post-LPS stimulation suggests that more time may be required following LPS treatment for IL-6 to elicit its effect on SAA3 expression. Finally, expression of CCL20, a CC-motif chemokine that is induced in response to E. coli mastitis (Petzl et al, 2012) and has been shown to differ in expression in 2 other groups of HR and LR fibroblast cohorts (Green et al, 2015;Benjamin et al, 2016), was measured. The chemokine was highly upregulated by 2 h post-LPS treatment and, overall, expression was higher in HR cells, but again the difference did not reach significance.…”

Section: Discussionmentioning

confidence: 99%

Variation in fibroblast expression of toll-like receptor 4 and lipopolysaccharide-induced cytokine production between animals predicts control of bacterial growth but not severity of Escherichia coli mastitis

Korkmaz

Elsasser

Kerr

2018

Journal of Dairy Science

View full text Add to dashboard Cite

Mastitis caused by environmental pathogens such as Escherichia coli is highly problematic to the dairy industry because it incurs substantial cost and tends to be difficult to manage. An effective innate immune response by the host is key to controlling infection, but it should also limit collateral damage to the mammary gland. Between-animal differences in mastitis severity have been attributed to variability in the innate response. In the current study, we used primary dermal fibroblast as a model to rank animals based on composite expression of the toll-like receptor 4 gene (TLR4) and lipopolysaccharide (LPS)-induced IL-8 and IL-6 protein production. Animals ranked as high and low responders (HR and LR, respectively) were then infected with the P4 strain of E. coli to determine how difference in rank would affect response to mastitis. All animals developed an acute response to the infection with varying degrees in severity; however, HR animals had an elevated somatic cell count and fever response at 12 h post-infection and greater production of milk IL-8 at 24 h post-infection. The HR animals were also significantly more capable of limiting bacterial growth. No differences in post-infection milk production or concentrations of milk BSA were measured. The current study indicates that HR animals have an early upregulation in their innate response that is beneficial for bacterial clearance; however, they are equally susceptible to tissue damage caused by an exuberant response to the infection. The dermal fibroblast may be used in conjunction with other cell types to determine how the innate response is regulated to mitigate unnecessary injury to the mammary gland while still effectively clearing the pathogen.

show abstract

“…Gene Regulation in Ruminants: A Nutritional Perspective DOI: http://dx.doi.org /10.5772/intechopen.82193 In a study aiming to use the fibroblast model to explore differences between a dairy breed (Holstein) and a beef breed (Angus) in their innate responses to LPS exposure, several immune-associated differentially expressed genes between breeds were found [124]. Within them, TLR4, which is the extracellular receptor responsible for recognition of LPS presented higher level of expression in Holstein cows as compared to Angus, suggests the Holstein animals will detect and respond to Gram-negative bacteria more vigorously than Angus animals.…”

Section: Example Of Nutrigenomics Linkage Between Beef and Dairy Cattlementioning

confidence: 99%

“…While a beef calf usually stays with the dam approximately until 6-7 months old, a dairy calf is separated from its mother as soon as it finishes consuming colostrum or earlier. Although there are studies that started to analyze the epigenetic differences between breeds [124], this is a promising area that needs to be studied in deep.…”

Section: Example Of Nutrigenomics Linkage Between Beef and Dairy Cattlementioning

confidence: 99%

Gene Regulation in Ruminants: A Nutritional Perspective

Osorio¹,

Moisá²

2019

Gene Expression and Control

View full text Add to dashboard Cite

This chapter will focus on cellular regulatory programs implemented by the ruminant physiology in order to respond to external stimuli such as nutrition as well as important physiological events such as parturition. The increasing adoption of "omics" technologies and bioinformatics in nutrition and physiology in ruminant research have allowed us to delineate a clearer picture on what regulates major biological process at a molecular level such as milk synthesis and meat quality and fatty acid composition as well as pathological conditions such as ketosis, mastitis, and heat stress. The assembly of such plethora of information in a blend among nutritional research, molecular biology, and novel tools to study the response of the genome to nutrition has led to emerging disciplines such as nutritional genomics or "nutrigenomics."

show abstract

Differential responsiveness of Holstein and Angus dermal fibroblasts to LPS challenge occurs without major differences in the methylome

Cited by 14 publications

References 42 publications

Genome-wide methylation analysis reveals differentially methylated loci that are associated with an age-dependent increase in bovine fibroblast response to LPS

Genome-wide methylation analysis reveals differentially methylated loci that are associated with an age-dependent increase in bovine fibroblast response to LPS

Variation in fibroblast expression of toll-like receptor 4 and lipopolysaccharide-induced cytokine production between animals predicts control of bacterial growth but not severity of Escherichia coli mastitis

Gene Regulation in Ruminants: A Nutritional Perspective

Contact Info

Product

Resources

About