Development of a bead-based multiplex assay to quantify bovine interleukin-10, tumor necrosis factor-α, and interferon-γ concentrations in plasma and cell culture supernatant

Sipka, Anja; Mann, Sabine; Babasyan, Susanna; Freer, Heather; Wagner, Bettina

doi:10.3168/jdsc.2021-0191

Cited by 8 publications

(4 citation statements)

References 27 publications

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“…It is difficult to compare these findings with previous works as in many studies, the induced inflammation in HepG2 cells was analysed through the expression of cytokines [ 45 , 46 ] or, when the production of the cytokine is quantified, it is done mostly using the ELISA technique and not the bead-based multiplex technology used here. These techniques are not comparable because of differences in reagents and recombinant proteins used as standards, and information on assay validation is missing from some commercially available ELISA [ 47 , 48 ]. Another difference that limits comparison is the sample matrix [ 47 ] as in other studies primary human and rat hepatocytes [ 49 ] or human [ 20 ] or rat serum [ 42 ] are used.…”

Section: Discussionmentioning

confidence: 99%

Colonic Coffee Phenols Metabolites, Dihydrocaffeic, Dihydroferulic, and Hydroxyhippuric Acids Protect Hepatic Cells from TNF-α-Induced Inflammation and Oxidative Stress

Sánchez-Medina

Redondo-Puente

Dupak

et al. 2023

IJMS

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Coffee presents beneficial health properties, including antiobesity effects. However, its effects on inflammation are controversial. Hydroxycinnamic acids are the main coffee phenolic bioactive compounds. In human bioavailability studies carried out with coffee, among the most abundant compounds found in urine and plasma were the colonic metabolites, dihydrocaffeic (DHCA), dihydroferulic (DHFA), and hydroxyhippuric (HHA) acids. To understand the hepato-protective potential of these three compounds, we tested whether treatment with realistic concentrations (0.5–10 µM) were effective to counteract inflammatory process and oxidative status induced by tumor necrosis factor α (TNF-α). First, we established a novel model of inflammation/oxidation using TNF-α and HepG2 cells. Afterwards, we evaluated the activity of DHCA, DHFA, and HHA against the inflammatory/oxidative challenge through the determination of the inflammatory mediators, interleukins (IL)-6, and IL-8 and chemokines, monocyte chemoattractant protein-1, and macrophage inflammatory protein-1, as well as the levels of biomarkers of oxidative stress, such as reactive oxygen species, reduced glutathione, and the antioxidant enzymes glutathione peroxidase and reductase. Results showed that all three compounds have a potential hepato-protective effect against the induced inflammatory/oxidative insult.

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Section: Discussionmentioning

confidence: 99%

Colonic Coffee Phenols Metabolites, Dihydrocaffeic, Dihydroferulic, and Hydroxyhippuric Acids Protect Hepatic Cells from TNF-α-Induced Inflammation and Oxidative Stress

Sánchez-Medina

Redondo-Puente

Dupak

et al. 2023

IJMS

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“…Serum amyloid A (SAA) concentrations were determined using a commercial analysis kit (Multispecies SAA, Tridelta Development Ltd.) with at least 1,000-fold dilution of all samples. Plasma cytokine concentrations (TNF-α, IFN-γ, and IL-10) were determined using a multiplex bead-based assay as previously described by Sipka et al (2022). For quality control of all in-house assays, low and high concentrations of controls were run on every plate.…”

Section: Blood Sampling and Analysismentioning

confidence: 99%

Lipopolysaccharide challenge following intravenous amino acid infusion in postpartum dairy cows: II. Clinical and inflammatory responses

Chandler

Westhoff

Sipka

et al. 2022

Journal of Dairy Science

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Amino acids (AA) are integral nutrients for a functioning immune system. Postpartum cows experience AA deficits early postpartum that may influence the response to immune activation. This study investigated the clinical and inflammatory responses to a systemic inflammatory stimulus after a 4-d intravenous (IV) AA infusion with a mix of essential and nonessential AA designed to ameliorate the estimated metabolizable protein deficit in early postpartum cows. Our objectives were (1) to describe the clinical and inflammatory response to an acute IV lipopolysaccharide (LPS) challenge in early postpartum cows, and (2) to compare these clinical and inflammatory responses between IV AA-treated and control cows. Cows (n = 14, 4 ± 1 d in milk) were continuously infused IV for 4 d in a matchedpair randomized controlled design and received 0.9% NaCl (CTRL) or IV AA (IVAA) to supply 1 g/kg of BW per day of combined essential and nonessential AA. After infusion ended, cows were challenged with IV LPS (0.0625 µg/kg of BW over 1 h), and serial blood samples were collected for complete blood cell counts and to quantify plasma cytokines and acute-phase proteins. Body temperature, heart rate, and respiratory rate were monitored for 24 h during challenge. During challenge, maximum body temperature was greater in IVAA (41.3 ± 0.20°C) than in CTRL (40.6 ± 0.19°C). In both groups, respiratory rate increased during the first 2 h following challenge, whereas heart rate first decreased over the first 2 h and then increased to reach a maximum at 4 h. Acute leucopenia occurred within 1 h of challenge in both groups before leukocytosis was observed at 24 h, with white blood cell counts returning to baseline values within 72 h. Plasma haptoglobin and serum amyloid A concentrations increased 3-fold and 4-fold in both groups and peaked at 48 and 24 h following challenge, respectively. Plasma concentra-tions of TNF-α and IL-10 increased within 1 h and peaked at 2 h following the start of challenge. Plasma IL-10 concentrations increased to a greater extent in CTRL compared with IVAA during challenge. Despite differences in IL-10 concentration, previous AA infusion did not alter the acute-phase protein response to LPS challenge. We conclude that AA infusion before systemic inflammatory challenge decreased the antiinflammatory response but did not alter concentrations of other systemic markers of inflammation.

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“…Culture supernatants were used to quantify the concentration of IL10, TNF, and IFNG in a bead-based multiplex assay as described 29 .…”

Section: Methodsmentioning

confidence: 99%

Inhibition of mTOR in bovine monocyte derived macrophages and dendritic cells provides a potential mechanism for postpartum immune dysfunction in dairy cows

Sipka

Chandler

Weichhart

et al. 2022

Sci Rep

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Dairy cattle experience a profound nutrient deficit postpartum that is associated with immune dysfunction characterized by heightened inflammation and reduced pathogen clearance. The activation of the central nutrient-sensing mTOR pathway is comparatively reduced in leukocytes of early postpartum dairy cows during this time of most pronounced nutrient deficit. We assessed the effect of pharmacological mTOR inhibition (Torin-1, rapamycin) on differentiation of monocyte derived classically (M1) and alternatively (M2) activated macrophages (MPh) and dendritic cells (moDC) from 12 adult dairy cows. Treatment with mTOR inhibitors generated M1 MPh with increased oxidative burst and expression of IL12 subunits but decreased phagocytosis and expression of IL1B, IL6, and IL10. In M2 MPh, treatment inhibited expression of regulatory features (CD163, ARG2, IL10) skewing the cells toward an M1-like phenotype. In moDC, mTOR inhibition increased expression of pro-inflammatory cytokines (IL12A, IL12B, IL1B, IL6) and surface CD80. In co-culture with mixed lymphocytes, mTOR-inhibited moDC exhibited a cytokine profile favoring a Th1 response with increased TNF and IFNG production and decreased IL10 concentrations. We conclude that mTOR inhibition in vitro promoted differentiation of inflammatory macrophages with reduced regulatory features and generation of Th1-favoring dendritic cells. These mechanisms could contribute to immune dysregulation in postpartum dairy cows.

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Development of a bead-based multiplex assay to quantify bovine interleukin-10, tumor necrosis factor-α, and interferon-γ concentrations in plasma and cell culture supernatant

Cited by 8 publications

References 27 publications

Colonic Coffee Phenols Metabolites, Dihydrocaffeic, Dihydroferulic, and Hydroxyhippuric Acids Protect Hepatic Cells from TNF-α-Induced Inflammation and Oxidative Stress

Colonic Coffee Phenols Metabolites, Dihydrocaffeic, Dihydroferulic, and Hydroxyhippuric Acids Protect Hepatic Cells from TNF-α-Induced Inflammation and Oxidative Stress

Lipopolysaccharide challenge following intravenous amino acid infusion in postpartum dairy cows: II. Clinical and inflammatory responses

Inhibition of mTOR in bovine monocyte derived macrophages and dendritic cells provides a potential mechanism for postpartum immune dysfunction in dairy cows

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