These data are also consistent with the hypothesis that THP-1 cells have more effective mechanisms for managing oxidative stress in comparison to Jurkat cells. 3.3. phagocytosis Anemoside A3 of NTP-exposed leukemic cells by macrophages. Our results highlight the significance of target-mediated modulation of plasma chemical species in the development and clinical use of protocols involving plasma sources for use in cancer therapeutic application. Abstract Non-thermal plasma application to cancer cells is known to induce oxidative stress, cytotoxicity and indirect immunostimulatory effects on antigen presenting cells (APCs). The purpose of this study was to evaluate the responses of two leukemic cell linesJurkat T lymphocytes and THP-1 monocytesto NTP-generated reactive oxygen and nitrogen species (RONS). Both cell types depleted hydrogen peroxide, but THP-1 cells neutralized it almost immediately. Jurkat cells transiently blunted the frequency-dependent increase in nitrite concentrations in contrast to THP-1 cells, which exhibited no immediate effect. A direct relationship between frequency-dependent cytotoxicity and mitochondrial superoxide was observed only in Jurkat cells. Jurkat cells were very responsive to NTP in their display of calreticulin and heat shock proteins 70 and 90. In contrast, THP-1 cells were minimally responsive or unresponsive. Despite no NTP-dependent decrease in cell surface display of CD47 in either cell line, both cell types induced migration of and phagocytosis by Anemoside A3 APCs. Our results demonstrate that cells modulate the RONS-mediated changes in liquid chemistry, and, importantly, the resultant immunomodulatory effects of NTP can be independent of NTP-induced cytotoxicity. 0.01, *** 0.001, **** 0.0001). Changes in NTP-dependent nitrite and hydrogen peroxide concentrations in the presence of THP-1 cells differed from those observed with Jurkat cells. At T0, increases in hydrogen peroxide concentrations were negligible or considerably diminished in the presence of THP-1 cells and were only modestly frequency-dependent (5.80 M at 30 Hz to 10.88 M at 105 Hz), with significant increases noted only at frequencies of 60 Hz (8.64 M) and higher (Figure 2a). Similarly, hydrogen peroxide concentrations at T24 were considerably lower in the presence of THP-1 cells (13.23 M compared to 59.44 M in RPMI only at 105 Hz) and not frequency-dependent. Nitrite concentrations in the presence of THP-1 cells (Figure 2b) were frequency-dependent at T0 (increasing from 6.09 M at 30 Hz to 20.00 M at 105 Anemoside A3 Hz) and T24 (increasing from 7.70 M at 30 Hz to 18.78 M at 105 Hz), similar between T0 and T24 and comparable to concentrations detected in media in the absence of THP-1 cells at T0 (5.95 M at 30 Hz to 23.63 M at 105 Hz in RPMI without cells). By T24, CALCA nitrite concentrations in the presence of THP-1 cells were still frequency-dependent but somewhat lower than concentrations measured in the absence of cells (18.78 M compared to 29.55 M at 105 Hz in RPMI without cells). These results show that NTP-associated peroxide and nitrite chemistries in media are cell type-dependent. 3.2. Lymphocytic and Myeloid Leukemic Cells Differ in Susceptibility to NTP Differences in peroxide and nitrite chemistry in media attributable to the presence of either cell type suggested that cellular responses to NTP exposure would also differ between Jurkat and THP-1 cells. To address this hypothesis, Anemoside A3 we measured the NTP-dependent cytotoxicity in both cell lines. Previous studies demonstrated that application of NTP to Jurkat cells resulted in increased oxidative stress and stimulation of pro-apoptotic pathways [26,27,28]. In agreement with these studies, we observed that exposure of Jurkat cells to nsDBD plasma induced a frequency-dependent decrease in cell viability, with considerable cytotoxicity observed at 105 Hz (Figure 3a). Consistent with the assumption that NTP-induced cytotoxicity is due to oxidative stress, pre-treatment with the antioxidant N-acetyl cysteine (NAC) ameliorated this cytotoxicity. Open in a separate.