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Box-and-whisker plot of the effect of the medium-preparation method and sample storage temperature during transportation on the interleukin 10 (IL10) response to positive control pokeweed mitogen (PWM) and Coxiella burnetii (CB) antigen stimulation. Whole blood from 6 cows was transported either on ice (green) or at ambient temperature (brown) then stimulated in vitro for 48 h with A ) PWM, or B ) CB phase 1 or 2. Cytokine response was determined using an ELISA. Bold black lines are medians; triangles are means; large dots are outliers. Note: y-axis scales for A and B are different.

Journal: Journal of Veterinary Diagnostic Investigation: Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

Article Title: Optimization of a bovine whole-blood cytokine recall assay for the detection of interferon-gamma and interleukin 10 following stimulation with Coxiella burnetii antigens

doi: 10.1177/10406387251390933

Figure Lengend Snippet: Box-and-whisker plot of the effect of the medium-preparation method and sample storage temperature during transportation on the interleukin 10 (IL10) response to positive control pokeweed mitogen (PWM) and Coxiella burnetii (CB) antigen stimulation. Whole blood from 6 cows was transported either on ice (green) or at ambient temperature (brown) then stimulated in vitro for 48 h with A ) PWM, or B ) CB phase 1 or 2. Cytokine response was determined using an ELISA. Bold black lines are medians; triangles are means; large dots are outliers. Note: y-axis scales for A and B are different.

Article Snippet: Microplates (96-well Nunc MaxiSorb) were coated with mouse anti-bovine IL10 antibody (clone CC318; Bio-Rad) at a concentration of 2.5 μg/mL diluted in wash buffer (0.05% Tween 20 in RO water) and incubated at 4°C overnight.

Techniques: Whisker Assay, Positive Control, In Vitro, Enzyme-linked Immunosorbent Assay

Box-and-whisker plot of the distribution of the interleukin 10 (IL10) response to Coxiella burnetii at various dilutions and incubation times in whole blood from 4 cows. Whole blood was stimulated in vitro with C. burnetii phases 1 (P1 CB) and 2 (P2 CB) at concentrations of 5 μg/mL, 0.5 μg/mL, or 0.05 μg/mL and incubated for 24 h (yellow) or 48 h (green). Other stimulants included lipopolysaccharide from rough-form Escherichia coli (LPS), medium (unstimulated control), and pokeweed mitogen (PWM; positive control). Cytokine response was determined using an ELISA. Bold black bars are medians; triangles are means; large dots are outliers. Cytokine response to PWM is presented on the secondary y-axis.

Journal: Journal of Veterinary Diagnostic Investigation: Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

Article Title: Optimization of a bovine whole-blood cytokine recall assay for the detection of interferon-gamma and interleukin 10 following stimulation with Coxiella burnetii antigens

doi: 10.1177/10406387251390933

Figure Lengend Snippet: Box-and-whisker plot of the distribution of the interleukin 10 (IL10) response to Coxiella burnetii at various dilutions and incubation times in whole blood from 4 cows. Whole blood was stimulated in vitro with C. burnetii phases 1 (P1 CB) and 2 (P2 CB) at concentrations of 5 μg/mL, 0.5 μg/mL, or 0.05 μg/mL and incubated for 24 h (yellow) or 48 h (green). Other stimulants included lipopolysaccharide from rough-form Escherichia coli (LPS), medium (unstimulated control), and pokeweed mitogen (PWM; positive control). Cytokine response was determined using an ELISA. Bold black bars are medians; triangles are means; large dots are outliers. Cytokine response to PWM is presented on the secondary y-axis.

Article Snippet: Microplates (96-well Nunc MaxiSorb) were coated with mouse anti-bovine IL10 antibody (clone CC318; Bio-Rad) at a concentration of 2.5 μg/mL diluted in wash buffer (0.05% Tween 20 in RO water) and incubated at 4°C overnight.

Techniques: Whisker Assay, Incubation, In Vitro, Control, Positive Control, Enzyme-linked Immunosorbent Assay

Box-and-whisker plot of the distribution of the interleukin 10 (IL10) response (ng/mL) of whole blood collected from 46 Australian dairy cows sampled at calving (yellow) and ~8 wk post-calving (blue). Whole blood was stimulated in vitro with Coxiella burnetii (CB) phase 1, lipopolysaccharide from rough-form Escherichia coli (LPS), pokeweed mitogen (PWM; positive control), or medium (unstimulated control). Cytokine response was determined using ELISA. Thick bars are medians; triangles are means; large dots are outliers. Cytokine response to PWM is presented on the secondary y-axis.

Journal: Journal of Veterinary Diagnostic Investigation: Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

Article Title: Optimization of a bovine whole-blood cytokine recall assay for the detection of interferon-gamma and interleukin 10 following stimulation with Coxiella burnetii antigens

doi: 10.1177/10406387251390933

Figure Lengend Snippet: Box-and-whisker plot of the distribution of the interleukin 10 (IL10) response (ng/mL) of whole blood collected from 46 Australian dairy cows sampled at calving (yellow) and ~8 wk post-calving (blue). Whole blood was stimulated in vitro with Coxiella burnetii (CB) phase 1, lipopolysaccharide from rough-form Escherichia coli (LPS), pokeweed mitogen (PWM; positive control), or medium (unstimulated control). Cytokine response was determined using ELISA. Thick bars are medians; triangles are means; large dots are outliers. Cytokine response to PWM is presented on the secondary y-axis.

Article Snippet: Microplates (96-well Nunc MaxiSorb) were coated with mouse anti-bovine IL10 antibody (clone CC318; Bio-Rad) at a concentration of 2.5 μg/mL diluted in wash buffer (0.05% Tween 20 in RO water) and incubated at 4°C overnight.

Techniques: Whisker Assay, In Vitro, Positive Control, Control, Enzyme-linked Immunosorbent Assay

Scatter plots of associations between cytokine response (ng/mL) from cattle whole blood stimulated in vitro with Coxiella burnetii (CB) phase 1 or phase 2 for 169 Australian dairy cows measured at ~8 wk post-calving: A ) interferon-gamma (IFNγ); B ) interleukin 10 (IL10).

Journal: Journal of Veterinary Diagnostic Investigation: Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

Article Title: Optimization of a bovine whole-blood cytokine recall assay for the detection of interferon-gamma and interleukin 10 following stimulation with Coxiella burnetii antigens

doi: 10.1177/10406387251390933

Figure Lengend Snippet: Scatter plots of associations between cytokine response (ng/mL) from cattle whole blood stimulated in vitro with Coxiella burnetii (CB) phase 1 or phase 2 for 169 Australian dairy cows measured at ~8 wk post-calving: A ) interferon-gamma (IFNγ); B ) interleukin 10 (IL10).

Article Snippet: Microplates (96-well Nunc MaxiSorb) were coated with mouse anti-bovine IL10 antibody (clone CC318; Bio-Rad) at a concentration of 2.5 μg/mL diluted in wash buffer (0.05% Tween 20 in RO water) and incubated at 4°C overnight.

Techniques: In Vitro

Scatter plots of associations between cytokine response to Coxiella burnetii (CB) phases 1 and 2 when presented as a sample:positive ratio (S:P ratio [%]) or in ng/mL for A, B ) interferon-gamma (IFNγ) and C, D ) interleukin 10 (IL10) from 169 dairy cows sampled at 8 wk post-calving. S:P ratio (%) calculated as: ([OD C. burnetii stimulation – OD medium background control]/[OD stimulation positive control (PWM) – OD medium background]) × 100%. Values in ng/mL were obtained by transforming OD values according to a standard curve and calculated as: ng/mL C. burnetii stimulation – ng/mL unstimulated control.

Journal: Journal of Veterinary Diagnostic Investigation: Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

Article Title: Optimization of a bovine whole-blood cytokine recall assay for the detection of interferon-gamma and interleukin 10 following stimulation with Coxiella burnetii antigens

doi: 10.1177/10406387251390933

Figure Lengend Snippet: Scatter plots of associations between cytokine response to Coxiella burnetii (CB) phases 1 and 2 when presented as a sample:positive ratio (S:P ratio [%]) or in ng/mL for A, B ) interferon-gamma (IFNγ) and C, D ) interleukin 10 (IL10) from 169 dairy cows sampled at 8 wk post-calving. S:P ratio (%) calculated as: ([OD C. burnetii stimulation – OD medium background control]/[OD stimulation positive control (PWM) – OD medium background]) × 100%. Values in ng/mL were obtained by transforming OD values according to a standard curve and calculated as: ng/mL C. burnetii stimulation – ng/mL unstimulated control.

Article Snippet: Microplates (96-well Nunc MaxiSorb) were coated with mouse anti-bovine IL10 antibody (clone CC318; Bio-Rad) at a concentration of 2.5 μg/mL diluted in wash buffer (0.05% Tween 20 in RO water) and incubated at 4°C overnight.

Techniques: Control, Positive Control