Evaluating the Mitochondrial Activity and Inflammatory State of Dimethyl Sulfoxide Differentiated PLB‐985 Cells
Notice bibliographique
Résumé
INTRODUCTION Neutrophils play a key role as the first line of defense against pathogens. Their ability to generate and release inflammatory mediators (eicosanoids or cytokines) helps promote the inflammatory response and consequently restore the hemostasis. While active participants in several steps of the normal inflammatory response, neutrophils are also involved in chronic inflammatory diseases such as asthma, atherosclerosis and arthritis. Given their dual role in the modulation of inflammation, regulating the inflammatory response of neutrophils has been an important therapeutic approach pursued by numerous researchers. The neutrophil's lifespan is relatively short (8–12h), which can be problematic for some in vitro experiments. To address this issue, researchers used the human monomyelocyte cell line PLB‐985 as an alternative for some of their in vitro experiments. PLB‐985 cells can be differentiated into a more neutrophil‐like phenotype upon exposure to several agonists, including dimethyl sulfoxide (DMSO). Whether this differentiation of PLB‐985 affects important features related to neutrophil's normal functions ( i.e . mitochondrial activity, eicosanoid production) remains elusive and characterizing these changes will be the focal point of this project. OBJECTIVES We assessed and compared the mitochondrial activity and inflammatory state of PLB‐985 cells following differentiation. METHODS Differentiation of PLB‐985 cells was performed using 1% DMSO for 6 days and was confirmed by immunofluorescence microscopy and flow cytometry using an anti‐CD11b‐PE cell surface antibody. Proliferation and apoptosis assays were performed by flow cytometry using the eFluor 670 and 7‐aminoactinomycin D labelers respectively. The mitochondrial respiratory assay was determined by high‐resolution O 2 respirometry (Oroboros Instrument). The inflammatory state of the cells was evaluated by transcriptomic, proteomic and lipidomic approaches. RESULTS Differentiation affected the proliferation of PLB‐985 cells ( P <0.0001), without inducing apoptosis. A significant decrease in the mitochondrial respiration was observed in differentiated PLB‐985 cells ( P <0.0005). However, the overall mitochondria content was not affected. Immunoblotting with mitochondrial antibodies revealed a strong modulation of the succinate dehydrogenase A ( P <0.0001), superoxide dismutase 2 ( P <0.0001), ubiquinol‐cytochrome c reductase core protein 2 ( P <0.05) and ATP synthase subunit α ( P <0.01) in differentiated PLB‐985 cells. Finally, eicosanoids (leukotriene B 4 , 12‐hydroxyheptadecatrienoic and 15‐hydroxyeicosatetraenoic acids) production was significantly increased in differentiated cells ( P <0.01). CONCLUSIONS Our data demonstrate that the differentiation process of PLB‐985 cells does not impact their viability despite a reduced respiratory state of the cells. This process is also accompanied by modulations of the inflammatory state of the cell. Finally, our data suggests that PLB‐985 cells could be a suitable in vitro candidate to study mitochondrial‐related dysfunctions in inflammatory diseases. Support or Funding Information CIHR, NBHRF, NBIF This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .
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 | 0,001 |
| 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 ».