Optimization of Capture ELISAs for Chicken Cytokines Using Commercially Available Antibodies

Krzysica, Paulina; Verhoog, Loes; Vries, S. de; Smits, C.H.M.; Savelkoul, H.F.J.; Tijhaar, Edwin

doi:10.3390/ani12213040

Cited by 6 publications

(4 citation statements)

References 27 publications

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“…Today, the commercially available repertoire of avian cytokines consists of about 25 elements, all of which were utilized by ELISA or Western blotting, and the list is still expanding ( Lu et al, 2023 ). For example, a recent study has calibrated an ELISA assay for chicken IL-2, IL-6, IL-10, IL-12p40, and IFN-γ ( Gonzalez et al, 2022 ). The antibodies reliably detected native cytokines and exhibited sensitivity as low as 32 pg/mL.…”

Section: Discussionmentioning

confidence: 99%

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A decline in avian cytokine expression with age revealed by commercially available cytokine array

et al. 2023

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Cytokines are secreted immunomodulators that are key regulators of the avian immune response. Currently, the most commonly used method to follow cytokine expression is qPCR, which measures cellular levels of mRNA, rather their extracellular circulating levels. Here we present a commercially available cytokine array designed to assay circulating expression levels of multiple cytokines and immunomodulators simultaneously. Upon minor modifications to the manufacturer protocol, background noise was reduced, leading to a significant increase in the sensitivity of the device. Our data indicate that the array is reliable and produce consistent data between biological repeats. We tested the reproducibility of the array in a biologically relevant context by assessing age-related changes in circulating cytokines. While individual features did not show a consistent pattern, our data revealed a consistent decline in the median of all cytokine values, supporting the validity of the array in studying biological processes.

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Section: Discussionmentioning

confidence: 99%

“…However, this method only measures cellular cytokine expression at the mRNA level and does not provide information about actual concentrations of circulating cytokines ( Gonzalez et al, 2022 ).…”

Section: Introductionmentioning

confidence: 99%

A decline in avian cytokine expression with age revealed by commercially available cytokine array

et al. 2023

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“…In the present study, the Merck Miliplex Cytokine Assay Kit (Cat GCYT1-16K. Millipore Corporation Merck, Darmstadt, Germany) was used according to the supplier’s instructions [ 22 ].…”

Section: Methodsmentioning

confidence: 99%

An Assessment of the Effects of Guanidinoacetic Acid on the Performance and Immune Response of Laying Hens Fed Diets with Three Levels of Metabolizable Energy

García-Gómora,

Gómez-Verduzco,

Márquez-Mota

et al. 2024

Animals

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Different levels of metabolizable energy (ME) and the inclusion of guanidinoacetic acid (GAA) in the diet of 53-week-old Lohmann LSL-CLASSIC hens were used to evaluate its effect on reproductive parameters, egg quality, intestinal morphology, and the immune response. Six diets were used in a 3 × 2 factorial design, with three levels of ME (2850, 2800, and 2750 kcal/kg), and with (0.08%) or without the inclusion of GAA. The addition of GAA to diets with low levels of ME increased (p < 0.05) egg production and egg mass. Moreover, hens fed with 2800 kcal/g without GAA had the highest concentration (p < 0.05) of serum interleukin IL-2, while those fed diets with the same amount of ME but supplemented with 0.08% GAA had the lowest concentration. Finally, the inclusion of 0.08% GAA increased (p < 0.05) the concentration of vascular endothelial growth factor (VEGF), regardless of the ME level in the diet. This study highlights the potential role of GAA in decreasing the energy level of ME (50–100 kcal/g) in the feeding of hens and in the modulation of specific immune responses. Further research is recommended to fully understand the mechanisms of action of GAA on the mechanism target of rapamycin and its relationship with the immune response.

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“…Total IgY observations were conducted using the Enzyme-Linked Immunosorbent Assay (ELISA) method as reported by Ruhi et al (2018). ELISA is widely applied in immunological research in poultry due to its ease and efficiency (Krzysica et al, 2022). In this ELISA test, egg yolk and blood serum of IPB-D3 chickens are used to determine the concentration of antibodies to the disease in IPB-D3 chickens.…”

Section: Total Igy In Egg Yolk and Chicken Serum Ipb-d3mentioning

confidence: 99%

Production Performance and Humoral Immunity in IPB-D3 Chickens

Indriyani,

Murtini,

Darwati

et al. 2024

AAVS

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This research were conducted analyzed antibodi in IPB-D3 chickens on production performance. Pedigree data and information were collected in generations 3 and 4. In the third generation, as many as 36 chickens (9 males and 27 females) aged 27 weeks and 162 IPB-D3 chicken eggs, in the fourth generation aged DOC and 2 weeks, each 81 samples. Total Immunoglobulin yolk (IgY) observations were made using Enzyme Linked Immunosorbent Assay (ELISA) and antibody titer against Newcastle disease virus (AbNDV) methods using Hemaglutination Inhibition (HI) tests. Descriptive analysis was performed on production performance data and humoral immunity to estimate phenotypic correlation of the statistical method of Pearson correlation analysis. The results of IgY examination in men aged 27 weeks were 9.49 ± 1.37 mg/mL and IgY in women 9.36 ± 2.89 mg/mL. AbNDV ages 0 and 2 weeks at 1.34±0.95 and 1,370.95 LOG 2 GMT. At the age of 12 weeks, the body weight of IPB-D3 chickens of G4 males is higher than that of G3 males, which is 1021.44±60.05 grams with a CV of 17.01%. Total egg production in a day in the age range of 29-40 weeks is 13.3±7.56 eggs, with a production percentage of 44.8%. Phenotypic correlation between IgY aged 0-2 with active body weight of IPB-D3 chickens aged 0 to 12 weeks showed positive but low to moderate values of 0,06-0,49. In conclusion, humoral immunity such as IgY and AbNDV did not have a significant effect on the growth and production of IPB-D3 chickens.

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Optimization of Capture ELISAs for Chicken Cytokines Using Commercially Available Antibodies

Cited by 6 publications

References 27 publications

A decline in avian cytokine expression with age revealed by commercially available cytokine array

A decline in avian cytokine expression with age revealed by commercially available cytokine array

An Assessment of the Effects of Guanidinoacetic Acid on the Performance and Immune Response of Laying Hens Fed Diets with Three Levels of Metabolizable Energy

Production Performance and Humoral Immunity in IPB-D3 Chickens

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