Large-Scale Production of Autologous CD14+-Monocyte Derived Dendritic Cells Co-Electroporated with Amplified Total Tumour mRNA and Human CD40L mRNA in Patients with B-Cell Chronic Lymphocytic Leukemia.
Notice bibliographique
Résumé
Abstract BACKGROUND: B-cell chronic lymphocytic leukemia (CLL) is currently incurable with conventional chemotherapeutic approaches. The induction of specific anti-CLL immune responses in vitro using autologous tumour-antigen loaded dendritic cell (DC)-based approaches has been previously demonstrated. The aim of this study was to optimize and validate large-scale production of an autologous DC vaccine prior to initiation of a planned Phase I/II clinical trial for patients with previously treated CLL. STUDY DESIGN: Following informed consent, 8 patients with confirmed CLL had 11 leukapheresis collections for mononuclear cells performed between April 2005 and May 2006. Methods for vaccine production were optimized during an initial cohort of CLL patients (N=7); production methods ‘at scale’ were subsequently confirmed with a validation cohort (N=3). METHODS: Leukemic B-cells were positively selected (CD19+/CD5+) and preserved in RNAlater™ prior to overnight shipment to Argos Therapeutics Inc (Durham, NC); amplified patient specific total tumour mRNA and huCD40L mRNA were subsequently supplied by Argos for final vaccine formulation. CD14+ monocytes were positively selected using a CliniMACS device (Miltenyi Biotech, Germany). Purified monocytes were subsequently cultured for 5 days in growth medium containing IL-4 and GM-CSF; DCs were matured with addition of TNF-β, IFN-γ, and PGE2 on day 5. DCs were harvested on day 6, co-electroporated with autologous total tumour RNA and huCD40L RNA, and cryopreserved for subsequent analysis. RESULTS: Mean patient age was 62 years, 6/8 were male, and all had Rai clinical stage I, previously untreated disease; mean peripheral white blood cell count was 17.2 × 109/L (range 7.4 – 26.7 × 109/L). Mean CD14+ monocyte yield post-CliniMACS selection was 2.42 × 109 cells (range 1.51 – 3.53 × 109) and monocyte purity was high (mean 96.5%, range 80 – 99.7%). In the validation cohort, day 6 immunophenotype (mean, range), measured 4 hours post-electroporation, was consistent with mature, activated DCs: CD14+ 2.5% (1.21–5.00), CD80+ 98.2% (98.0–99.7), CD83+ 87.5% (78.0 – 94.7), and CD86+ 99.5% (99.0–99.9). Mean CD40L expression, a surrogate marker for electroporation efficiency, was 80.3% (range 70.2 – 88.0); mean CD209 expression was 97.7% (95–99.7). Absolute numbers of DCs generated post-electroporation ranged from 1.26 × 108/L to 3.24 × 108/L and post-thaw DC viability ranged from 71–94%. T-cell co-culture experiments confirmed the generation of specific, autologous cytotoxic T-lymphocyte (CTL) responses to CLL targets, demonstrated by: (1) MHC class I restricted autologous CLL induced INF-γ response (intracellular staining), and (2) statistically significant functional MHC class I restricted CTL response (chromium-release assay). CONCLUSIONS: This study confirms the feasibility of generating large numbers of autologous CD14+-derived dendritic cells co-electroporated with patient specific total tumour RNA and huCD40L RNA from CLL patients. These data provide justification for a currently accruing Phase I/II clinical trial designed to evaluate this treatment in CLL patients that have a stable, low disease burden following at least one course of systemic chemotherapy.
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 | 0,000 |
| 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) | 0,001 | 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 ».