What's Normal? Using Standardized Flow Cytometry Immunophenotyping to Create Comprehensive Pediatric Reference Ranges
Notice bibliographique
Résumé
Studying pediatric patients includes challenges such as difficulty obtaining samples, limited normal control reference data and small available sample volumes. Standardized flow cytometry immunophenotyping panels are commercially available to explore lymphocyte markers and extensive normal adult ranges have been published by the ONE Study group; comparable pediatric control data do not exist. Our objective is to establish a reference dataset to be available for pediatric studies using standardized, comprehensive flow cytometry panels with fresh whole blood. Our group had the opportunity to collaborate with the local clinical hematology laboratory to obtain normal pediatric samples with the goal of establishing comprehensive reference ranges across the pediatric age spectrum. This study also acted as a trial of this methodology for potential implementation in the clinical laboratory. Methods Using 700uL of EDTA blood, DuraClone IM (Beckman Coulter) flow cytometry phenotyping was performed on healthy pediatric controls ranging from 66 days to 16 years of age (n=10). Samples were tested in five, 10-colour flow cytometry T and B cell panels. Acquisition was performed on a Navios cytometer following calibration and standardization of cytometer output by Flow-Set™ Pro Fluorospheres. Analysis of the markers programmed cell death protein 1 (PD-1), often noted as a measure of T cell exhaustion, and gamma/delta (γδ) T cells and their associated markers Vd1 and Vd2, was performed. Results Preliminary analyses show age related trends in PD-1 + T cells with increasing PD-1+ CD4 T cells with increasing age as shown in Figure 1 (R2=0.701); this association is much weaker for CD8 T cells. The number of CD3+ T cells, including γδ T cells, decreases with age, while the % of each population of cells remains constant. There is a trend indicating a decreasing ratio of V delta 1 to V delta 2 γδ T cells with age. Conclusion Duraclone is a rapid immunophenotyping method that requires small blood volumes. These results begin to establish valuable reference data for pediatric transplant patient populations and suggest that these flow panels show promise for application in the clinical flow cytometry laboratory due to their standardized nature. We will continue to explore age-related trends as we analyse additional data from these controls and test additional samples. Complete blood count data are available to calculate absolute cell counts and this analysis will be performed. The use of fresh whole blood in this project differs from published pediatric reference range studies utilizing frozen cells and ensures populations are not lost or altered during mononuclear cell isolation or freeze thaw cycles. This study highlights the value of partnership between research and clinical laboratories as it can facilitate access to otherwise difficult to obtain samples and is valuable to the clinical laboratory in the exploration of new methodologies. Women and Children's Health Reseach Institute (WCHRI).
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
Comment cette classification a été obtenuedéplier
Prédiction machine sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Le volet Gemma est une étiquette directe du modèle pour chaque travail de la base, lue sur la notice réduite au titre. Le volet Codex est un classifieur appris des 10 348 étiquettes directes de Codex et calibré sur les taux pondérés de l'échantillon; les champs sans appui suffisant ne portent aucun appel Codex. Le mode candidate est l'union des deux volets; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont pas des étiquettes humaines.
Scores du classifieur distillé par catégorie (deux têtes)
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,008 | 0,015 |
| Méta-épidémiologie (sens strict) | 0,001 | 0,000 |
| Méta-épidémiologie (sens large) | 0,001 | 0,000 |
| Bibliométrie | 0,003 | 0,002 |
| Études des sciences et des technologies | 0,001 | 0,001 |
| Communication savante | 0,002 | 0,001 |
| Science ouverte | 0,001 | 0,001 |
| Intégrité de la recherche | 0,001 | 0,001 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,002 | 0,001 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule source (Gemma direct ou Codex distillé), pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».