Analysis of the interaction between  T cells and glioblastoma cell line -derived microvesicles

ROSATO, MICAELA; SHIROMIZU, CAROLINA MAIUMI; ROSSO, DAVID ANTONIO; MELILLO, NADIA CAROLINA; INFANTE CRUZ, ALEJANDRA; SALAMONE, GABRIELA VERÓNICA; JANCIC, CAROLINA CAROLINA

Mar del Plata

Congreso; Reunión de Sociedades de Biociencias 2022. LXX Reunión Anual de la Sociedad Argentina de Inmunología y 3er Congreso Franco-Argentino de inmunología; 2022

Glioblastoma multiforme (GBM) is the most aggressive malignant cerebral tumor in adults and has a median survival of less than a year after diagnosis. GBM is refractory to standard treatments because of its infiltration nature. GBM immunotherapy based on γδ T cells has been proposed to treat this pathology. γδ T cells are non-conventional T lymphocytes that recognize malignant cells and can trigger their apoptosis. Furthermore, the tumor cells can secrete extracellular vesicles (EV), among them, microvesicles (MV), containing molecules that regulate tumor microenvironment to allow their growth and progression. The interplay between EV and target cells can be mediated by membrane receptors, and by the internalization or fusion of EV with cells. Previously, we demonstrated that MV released by GBM cell line U373 activated γδ T lymphocytes. In this work, we aimed to analyze the interaction between MV and γδ T cells. For that, peripheral blood γδ T lymphocytes were purified by using an anti-TCR γδ MicroBead isolation kit, and MV were obtained from U373 cell line conditioned medium by differential centrifugation. MV were then stained with the lipophilic fluorescent dye PKH26, and incubated with γδ T cells for 30 and 120 min. Afterward,the fluorescence on γδ T lymphocytes was evaluated by flow cytometry (n=4) and confocal microscopy (n=5). The analysis of γδ T cells by flow cytometry showed that after 30 min of incubation they were PKH26+ (p