Fig. 3.

TCR-NK-92 acquire T-cell like behaviors in terms of metabolic functions, morphodynamics and immune synapse.

(a-c) Mitochondrial respiration. Oxygen Consumption Rate (OCR) of NK-92, CD3-NK-92, TCR(Radium-1)-NK-92, J6, J76 and J Radium-1 over time, in the presence or not of anti-CD3 (aCD3). Mitochondrial respiration function (expressed as OCR rate) was measured, first at baseline and, then, probed by the serial addition of oligomycin, FCCP and antimycin-A/rotenone (anti-A + rot). (a) Basal respiration, (b) ATP production and (c) maximal respiration capacity measured in NK-92, CD3-NK-92, TCR(Radium-1)-NK-92, J6, J76 and J Radium-1, either CD3 treated or untreated. (n = 21). Error bars represent SD. (d) ExtraCellular Acidification Rate (ECAR) of NK-92, CD3-NK-92, TCR(Radium-1)-NK-92, J6, J76 and J Radium-1, either CD3 treated or untreated, before and after addition of oligomycin is shown. Comparison of basal glycolytic activity between effector cells, either CD3 treated or untreated. (n = 21). Error bars represent SD. (e) Representative Reflection Interference Contrast (RIC) and TIRF micrographs of lymphocyte adhesion (RICM), actin and CD3 organization obtained after 20 min of spreading on glass slides coated with anti-HLA class I or -CD3. Scale bar 5 μm. (f) Box plot of contact area as determined from segmentation of RICM images. (g) Box plot of actin centrality. (h) Box plot of CD3 intensity at the cell-surface contact zone. For all box plots, n ≥ 50 cells per condition. (i) Confocal images of NK-92, TCR-NK-92 and T-cell previously activated and marked with anti Vβ3 (green) and phalloidin (red). Scale bar 4 μm. (j) Box plot of the anti Vβ3 signal intensity (n = 25). Error bars represent SD. (k) Box plot of the centralization of TCR molecules (cSMAC-number) (n = 25). Error bars represent SD. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)