Chapter 4 │ Page 139 (1 vs 0,77; p = 0.094), this was not captured in the immediate CD122 and CD73 expression profile (Figure 7a-c, right panel). Nevertheless, a dose-dependent decrease in CD112 was observed 24h after treatment, thus suggesting a downstream, stress-induced cellular response in target downregulation, rather than instant oxidative e ects of the applied NTP in this cell line. 3.5. NTP Stimulates The Expression Of The Activating NK Ligand MICA/B Lastly, to further evaluate the delicate balance between activating/inhibiting signals on NK cells following NTP treatment, we evaluated the influence of NTP on the activating NK ligand MICA/B. Our data showed that oxidation-induced e ects, immediately after application, were limited (Figure 8). Ligand expression decreased by 28% (1 vs 0.72; p = 0.0415) after 500Hz application in the Cal27 cell line, while these e ects were not measured in the other two cell lines. Figure 8: NTP stimulates the expression of the activating NK ligand MICA/B 24h post NTP treatment. HNSCC cell lines were treated with di erent regimes of NTP, and expression of the MICA/B ligands was analyzed with flow cytometry. The amount of MICA/B expression on the cell surface is represented as mean fluorescence intensity minus FMO control, normalized to untreated controls (normalized (Norm) ΔMFI), immediate (0h) and one day (24h) post-treatment. Data are represented as mean ± SEM, with individual values shown (n = 5). Statistical significance between untreated cells and the treated conditions was determined using the generalized linear mixed model with post hoc Dunnett’s test (* p ≤ 0.05; ** p ≤ 0.01). Outliers were calculated with the Grubbs’ Test. A A A B CB B B EF I MN N TU N N N H H BC B
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