Analysis of indoleamine 2,3-dioxygenase 1 (IDO1) expression of cultured cord blood adherent mononuclear cells as an indicator of atopic risk
Bibliographic record
Abstract
Maternal atopy is a known risk factor for allergy development in children. This link can be studied to find potential indicators of atopic risk by examining umbilical cord blood. Indoleamine 2 , 3-dioxygenase 1 ( IDO1 ), the initiator of the IDO pathway, plays a regulatory role in the immune response and may differ in expression in the adherent mononuclear cells (AMNC) of atopic and non-atopic individuals. Supernatants of these AMNC cultures may also exhibit different cytokine profiles. Cord blood samples were collected from consenting women undergoing elective Caesarian-sections and atopic status was self-reported. Mononuclear cells were isolated and cryopreserved. 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 ). 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 7.5x10 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. Supernatant cytokine level change relative to plain for Th2 cytokines IL-4 (A), IL-5 (B) and IL-13 (C). Error bars represent the standard error of the mean. Cultures of atopic and non-atopic AMNCs were plated at 5x10 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, supernatants were collected and analyzed. A=high atopic risk, NA=low atopic risk. Each condition/atopic risk group contains a minimum of 6 samples. Supernatant cytokine level change relative to plain for pro- and anti-inflammatory cytokines TNF-α (A), IL-6 (B) and IL-10 (C). Error bars represent the standard error of the mean. Cultures of atopic and non-atopic AMNCs were plated at 5x10 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, supernatants were collected and analyzed. A=high atopic risk, NA=low atopic risk. Each condition/atopic risk group contains a minimum of 6 samples. Preliminary differences detected suggest that further research could elucidate a suitable biomarker to predict atopic risk. 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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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.001 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".