Tracking Phagosome-Derived Vesicles in Macrophages withMicrofabricated Microparticles
Notice bibliographique
Résumé
Phagocytosis is a process by which a cell such as macrophage internalizes a micrometre-sized particle.It plays critical roles in immunity, tissue remodelling and repair.[1] Phagocytosis is characterized by the internalization of the particle in a vacuole called phagosome inside the cell.Formation, maturation and resolution of the phagosome is a complex process and is not well understood.Among this process is the appearance and trafficking of intracellular vesicles that are apparently derived from the phagosome and named phagosome-derived vesicles (PDVs).Existing techniques for tracking PDVs rely on using particles that can be enzymatically degraded into soluble fluorescent fragments inside the phagosome.These techniques are incapable of detecting whether components of the phagosome membrane are in the PDVs and tracking PDVs formed from phagosomes containing non-degradable particles.The objective of this study is to develop a novel method for tracking PDVs derived from membranous components phagosomes containing non-degradable particles.The method is featured by using microparticles fabricated with a microfabrication technique.The technique, which is based on integrating spin-coating and microcontact printing, allows a precise control on the size and composition of the microparticles.[2]The microparticles are loaded with a dialkylcarbocyanine dye for staining the membrane of the PDVs.In this study, we have established a protocol for fabricating the microparticles with a temperature-sensitive polymer.The microparticles can be readily phagocytosed by a commonly used model macrophage cell line.By using a fluorescent-temperature sensitive polymer, we have confirmed that the dialkylcarbocyanine dye can stain intracellular vesicles outside the phagosomes while the phagosomes maintain their membrane integrity.Moreover, most of the dye-stained vesicles are formed after the microparticles have been phagocytosed, indicating that the dye-stained vesicles are PDVs.Additionally, we have made the following findings about the dye-stained vesicles.(1) They can move longitudinally along the pseudopodia of the macrophages and tend to accumulate in the pseudopodia.(2) They are acidic as revealed by Lysoview staining.(3) They are co-localized with vesicles stained by a different dialkylcarbocyanine dye added in the cell-culture medium.(4) They are colocalized with vesicles stained by a fluorescent dextran added into the cell-culture medium.(5) They are colocalized with zymosan bioparticles that are later phagocytosed by the macrophages.These findings indicate that the PDVs can be formed from phagosome containing non-degradable particles; the PDVs contain membranous components of the phagosomes; and the PDVs allow material transfer between phagosomes within a macrophage.In addition to the model macrophages, this technique can, in principle, be used to track PDVs in other types of phagocytes such as microglia, which play a critical role in the pathology of Alzheimer's disease.
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 | 0,001 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 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 ».