Phagocytosis, the process by which cells engulf and eliminate foreign particles or pathogens, is a critical mechanism used by immune cells, particularly macrophages and dendritic cells. Assessing phagocytic activity serves as a valuable marker for immune cell function in the context of anticancer immunity due to its role in clearing cancer cells, debris, and promoting an anti-tumour immune response [30 (link),36 (link)]. Lipopolysaccharide (LPS) was used to induce phagocytosis in PBMCs. LPS (Sigma-Aldrich, Rehovot, Israel) was introduced into three wells of a 96-well dark plate, with each well receiving 10 µL of a 5 µg/mL solution. Negative controls were established in another three wells using 10 µL of LPS diluent. The final three wells acted as media controls and contained only RPMI and 10 µL of 5 µg/mL LPS without cells. After 2 h of incubation and activation of phagocytic function, the dark plate was subjected to centrifugation and the supernatant was then removed. Subsequently, 100 µL of pHrodo Green Zymosan Bioparticles (Invitrogen, Eugene, OR, USA) [37 ] was added to each well, followed by incubation at 37 °C for one hour. Fluorescence measurements were performed using a fluorimeter (Fluoroskan Ascent, Thermo Fisher Scientific, Waltham, MA, USA), with excitation set at 510 nm and detection at 538 nm.
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