82P Obtaining microvesicles from T-lymphocytes using various methods of induction
نویسندگان
چکیده
Microvesicles (MVs) are a subset of extracellular vesicles secreted by most body cells. They being actively studied as cell-free biosimilar drugs. Many ways inducing MVs investigated - natural (passive secretion, cell activation) and stressful (hypoxia, chemical impact, etc.). The culture T-lymphocytes was activated CD3/CD28 antibodies with subsequent expansion. Some the cells were reactivated. Cytochalasin B (CHB) used to chemically induce formation. Ultrasound (US) physical inductor. resulting purified differential centrifugation. samples immunoblotting nanoparticle tracking analysis (NTA). NTA method determine concentration (for all samples, about 108 particles/mL) dispersity particle size. induction activation antibodies, treatment CHB or US increased monodispersity particles without affecting their average application led decrease in appearance fraction larger (up 700nm). As result Western blot analysis, an increase level CD3 MVs, well induced US, revealed compared control. In obtained content granzyme detected. CHB, these proteins not With equal amount total protein, beta-actin noted series cells>natural MVs>USinduction>CHB induction. This observation can be explained change cytoplasm/membrane ratio due difference size (10μm 0.1μm, respectively), mechanism action CHV (inhibition actin filament synthesis). All methods induction, except for make it possible obtain carrying functionally significant molecules T-lymphocytes. Further study structure functions is needed. work supported Russian Science Foundation grant 19-74-20026, support Strategic Academic Leadership Program Kazan (Volga Region) Federal University (PRIORITET- 2030).
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ژورنال
عنوان ژورنال: Annals of Oncology
سال: 2022
ISSN: ['0923-7534', '1569-8041']
DOI: https://doi.org/10.1016/j.annonc.2022.09.083