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Analysis of indoleamine 2 , 3-dioxygenase 1 ( IDO1 ) expression of cultured cord blood adherent mononuclear cells as an indicator of atopic risk

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Once thawed, AMNCs were cultured and stimulated with interferon-gamma (IFN-γ 1μg/ml or 1ng/ml) with or without control standard endotoxin (CSE 10ng/ml)... In each condition, 7.5x10 cells were seeded for gene analysis and 5x10 cells were seeded for cytokine analysis... Cells were lysed for RNA isolation, reverse transcribed and cDNA levels were analyzed using qPCR... Supernatant cytokine levels were analyzed using the Luminex xMAP Technology... IDO1 expression was significantly increased in all stimulated conditions (P<0.05) except for the CSE only condition... The high atopic risk group displayed trend towards decreased IDO1 expression, however, high and low atopic risk groups did not show significant differences (Figure 1)... Supernatant cytokine analysis show heightened levels of Th2 cytokines IL-4, IL-5, IL-13 (Figure 2)... Similarly, heightened levels of TNF-α and IL-6 were observed, while levels of IL-10 were decreased in the high atopic risk samples in all stimulated conditions (Figure 3)... Due to the lack of significant differences between high and low atopic risk groups for IDO1 expression and cytokine expression, a reliable biomarker was not determined in this study.

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IDO1 gene expression fold changes relative to plain media control. IDO1 expression levels were normalized to HPRT1 expression. The error bars represent the standard error of the mean. Numbers per stimulation group are as indicated beneath the graph. Cultures of atopic and non-atopic AMNCs were plated at 7.5x106 cells per condition. Following 5.5 hours incubation with either plain media, 1 μg/ml IFN-γ, or 1 μg/ml IFN-γ and 10 ng/ml CSE, cells were lysed for RNA extraction. RNA was reverse transcribed and cDNA levels were analyzed.
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Figure 1: IDO1 gene expression fold changes relative to plain media control. IDO1 expression levels were normalized to HPRT1 expression. The error bars represent the standard error of the mean. Numbers per stimulation group are as indicated beneath the graph. Cultures of atopic and non-atopic AMNCs were plated at 7.5x106 cells per condition. Following 5.5 hours incubation with either plain media, 1 μg/ml IFN-γ, or 1 μg/ml IFN-γ and 10 ng/ml CSE, cells were lysed for RNA extraction. RNA was reverse transcribed and cDNA levels were analyzed.

Mentions: IDO1 expression was significantly increased in all stimulated conditions (P<0.05) except for the CSE only condition. The high atopic risk group displayed trend towards decreased IDO1 expression, however, high and low atopic risk groups did not show significant differences (Figure 1). Supernatant cytokine analysis show heightened levels of Th2 cytokines IL-4, IL-5, IL-13 (Figure 2). Similarly, heightened levels of TNF-α and IL-6 were observed, while levels of IL-10 were decreased in the high atopic risk samples in all stimulated conditions (Figure 3).


Analysis of indoleamine 2 , 3-dioxygenase 1 ( IDO1 ) expression of cultured cord blood adherent mononuclear cells as an indicator of atopic risk
IDO1 gene expression fold changes relative to plain media control. IDO1 expression levels were normalized to HPRT1 expression. The error bars represent the standard error of the mean. Numbers per stimulation group are as indicated beneath the graph. Cultures of atopic and non-atopic AMNCs were plated at 7.5x106 cells per condition. Following 5.5 hours incubation with either plain media, 1 μg/ml IFN-γ, or 1 μg/ml IFN-γ and 10 ng/ml CSE, cells were lysed for RNA extraction. RNA was reverse transcribed and cDNA levels were analyzed.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4125990&req=5

Figure 1: IDO1 gene expression fold changes relative to plain media control. IDO1 expression levels were normalized to HPRT1 expression. The error bars represent the standard error of the mean. Numbers per stimulation group are as indicated beneath the graph. Cultures of atopic and non-atopic AMNCs were plated at 7.5x106 cells per condition. Following 5.5 hours incubation with either plain media, 1 μg/ml IFN-γ, or 1 μg/ml IFN-γ and 10 ng/ml CSE, cells were lysed for RNA extraction. RNA was reverse transcribed and cDNA levels were analyzed.
Mentions: IDO1 expression was significantly increased in all stimulated conditions (P<0.05) except for the CSE only condition. The high atopic risk group displayed trend towards decreased IDO1 expression, however, high and low atopic risk groups did not show significant differences (Figure 1). Supernatant cytokine analysis show heightened levels of Th2 cytokines IL-4, IL-5, IL-13 (Figure 2). Similarly, heightened levels of TNF-α and IL-6 were observed, while levels of IL-10 were decreased in the high atopic risk samples in all stimulated conditions (Figure 3).

View Article: PubMed Central - HTML

AUTOMATICALLY GENERATED EXCERPT
Please rate it.

Once thawed, AMNCs were cultured and stimulated with interferon-gamma (IFN-γ 1μg/ml or 1ng/ml) with or without control standard endotoxin (CSE 10ng/ml)... In each condition, 7.5x10 cells were seeded for gene analysis and 5x10 cells were seeded for cytokine analysis... Cells were lysed for RNA isolation, reverse transcribed and cDNA levels were analyzed using qPCR... Supernatant cytokine levels were analyzed using the Luminex xMAP Technology... IDO1 expression was significantly increased in all stimulated conditions (P<0.05) except for the CSE only condition... The high atopic risk group displayed trend towards decreased IDO1 expression, however, high and low atopic risk groups did not show significant differences (Figure 1)... Supernatant cytokine analysis show heightened levels of Th2 cytokines IL-4, IL-5, IL-13 (Figure 2)... Similarly, heightened levels of TNF-α and IL-6 were observed, while levels of IL-10 were decreased in the high atopic risk samples in all stimulated conditions (Figure 3)... Due to the lack of significant differences between high and low atopic risk groups for IDO1 expression and cytokine expression, a reliable biomarker was not determined in this study.

No MeSH data available.


Related in: MedlinePlus