Radosević K, Schut T C, van Graft M, de Grooth B G, Greve J
University of Twente, Department of Applied Physics, Enschede, Netherlands.
J Immunol Methods. 1993 May 5;161(1):119-28. doi: 10.1016/0022-1759(93)90203-j.
This study demonstrates that it is possible to investigate the membrane potential of interacting cells during the cytotoxic process using flow cytometry. Changes in the membrane potential of NK and K562 cells, involved in a cell-mediated cytotoxic process, were studied by standard and slit-scan flow cytometry, using the membrane potential sensitive fluorescent probe DiBAC4(3). The NK cells were labeled with a membrane marker (TR-18 or DiI) prior to incubation with K562 cells and the conjugates that were formed could be identified on the basis of the membrane marker fluorescence and light scattering signals. With a slit-scan technique we measured the membrane potential of each cell in a conjugate separately. The results show that depolarization of the K562 cell occurs as a consequence of the cytotoxic activity of the NK cell. This depolarization appears to be an early sign of cell damage because the cell membrane still remains impermeable to propidium iodide. Our data also indicate that depolarization of the NK cell occurs as a result of its cytotoxic activity.
本研究表明，使用流式细胞术可以在细胞毒性过程中研究相互作用细胞的膜电位。通过标准和狭缝扫描流式细胞术，使用膜电位敏感荧光探针DiBAC4(3)，研究了参与细胞介导的细胞毒性过程的NK细胞和K562细胞的膜电位变化。在与K562细胞孵育之前，用膜标记物（TR-18或DiI）标记NK细胞，并且可以基于膜标记物荧光和光散射信号鉴定形成的共轭物。使用狭缝扫描技术，我们分别测量了共轭物中每个细胞的膜电位。结果表明，K562细胞的去极化是NK细胞细胞毒性活性的结果。这种去极化似乎是细胞损伤的早期迹象，因为细胞膜对碘化丙啶仍然不可渗透。我们的数据还表明，NK细胞的去极化是其细胞毒性活性的结果。
