Effects of ultrasound on apoptosis induced by anti-CD20 antibody in CD20-positive B lymphoma cells

Aim esent study was conducted to examine the thermal and non-thermal effects of ultrasound on apoptosis induced by anti-CD20 monoclonal antibody (rituximab). als and methods -4 cells, a CD20-positive cell line derived from B cell lymphomas with a BCL2 gene rearrangement, were exposed to continuous 1 MHz ultrasound for therapeutic use under an air- or CO2-saturated condition to control cavitation. Early apoptosis (EA) and secondary necrosis (SN) were examined by flow cytometry. Cavitation was determined by detecting the hydroxyl radicals derived from pyrolysis of water molecules using electron paramagnetic resonance-spin trapping. To assess thermal effects, cells were treated in a temperature-controlled water bath. s was a significant additive increase in EA and EA + SN observed in cells treated with rituximab combined with heat at 42 °C or non-thermal ultrasound at 0.5 W/cm2 under an air-saturated condition, where heat or ultrasound induced some cell death. A significant synergistic increase in EA and EA + SN was observed in cells treated with rituximab and ultrasound at 2.5 W/cm2 under CO2-saturated conditions, where inertial cavitations were completely suppressed. No enhancement was observed at a temperature less than 40 °C or ultrasound at 0.5 W/cm2 under CO2-saturated conditions. sion results suggest that the immuno-therapeutic application of ultrasound at relatively high-intensities combined with rituximab thus produces synergistic effects under conditions where the non-thermal and non-cavitational effects are predominant.