Abstract PO-096: Cytokines in the lumen of exosomes are undetectable by immunoassays distorting cytokine profiles in HNC patients and healthy donors
Notice bibliographique
Résumé
Abstract Background: Exosomes, now called small extracellular vesicles (sEV), play a key role in cell-to-cell signaling. They are produced by all cells, circulate freely and are present in all body fluids. Evidence indicates that biologically active cytokines are present on the surface and/or in the lumen of sEV. The contributions of intra-vesicular cytokines to cytokine levels in plasma of cancer patients are unknown. Methods: sEV were isolated by ultrafiltration/size exclusion chromatography from pre-cleared plasma obtained from 30 patients with head and neck squamous cell carcinoma (HNSCC) and 10 healthy donors (HDs). Multiplex immunoassays were used to measure cytokine levels in paired untreated and detergent treated (0.5% Triton X-100) plasma samples and in isolated detergent-treated sEV. Results: The presence of cytokines on the surface and lumen of sEV isolated from plasma of the patients and HDs was first confirmed by immunoblots and on-bead flow cytometry. sEV-associated cytokines were functional in various in vitro coincubation assays with responder immune or tissue cells. Using Ab microarrays for 80 cytokines, we showed that sEV from HNSCC plasma contained a greater variety of cytokines (71/80; 88.8%) than sEV of HDs (43/80; 53.8%). Patients’ sEV had higher cytokine levels (range p<0.01 to 0.05) than did sEV from HD’s plasma. A highly sensitive Curiox 65-plex platform was then used to compare cytokine levels in paired untreated and detergent-treated plasma of HNSCC patients. 51/65 (78.5%) cytokines in the panel were consistently detected in treated and untreated plasma, and the majority of these (45; 88.2%) were present at significantly higher levels (range p<0.0001-0.05) in detergent-treated plasma. The comparison of cytokine levels between treated plasma, untreated plasma and paired sEV showed that the observed differences were due to cytokines sequestered in sEV. The cytokines released from detergent-treated EVs significantly contributed to the incomplete overall cytokine profile of untreated plasma in HNSCC patients. Further, only cytokines in sEV or in detergent-treated plasma correlated with disease stage in HNSCC. Conclusions: In untreated patient plasma, multiplex immunoassays detected only soluble cytokines and missed cytokines contained in the sEV lumen. Permeabilization of sEV was necessary for the Ab-based detection of total plasma cytokines (i.e., soluble + sEV-bound). As sEV-associated cytokines are biologically active, their absence from the plasma cytokine profiles that are determined in untreated plasma leads to inaccurate cytokine measurements. Underestimated cytokine levels in disease likely contribute to the lack of clinically significant correlations of plasma cytokine levels with disease progression. Citation Format: Chang-Sook Hong, Brenda Diergaarde, Theresa L. Whiteside. Cytokines in the lumen of exosomes are undetectable by immunoassays distorting cytokine profiles in HNC patients and healthy donors [abstract]. In: Proceedings of the AACR-AHNS Head and Neck Cancer Conference: Innovating through Basic, Clinical, and Translational Research; 2023 Jul 7-8; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2023;29(18_Suppl):Abstract nr PO-096.
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 | 0,000 | 0,001 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,002 | 0,000 |
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 ».