Changes in Intracellular Calcium Concentration and pH of Target Cells During the Cytotoxic Process: A Quantitative Study at the Single Cell Level

B.G. de Grooth, Katarina Radosevic, Jan Greve, K. Radosevic

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    Abstract

    This study reports on the changes in intracellular calcium concentration ([Ca2+]in) and intracellular pH ([pH]in) that occur in K562 target cells during interaction with human Natural Killer (NK) cells. The data were obtained using a quantitative fluorescence microscope and fluorescent ratio probes specific for [Ca2+]in (Fura-2-AM) and [pH]in (BCECFAM). Results demonstrate that two types of target cell response to the attack by an NK cell can be distinguished. The target cell either dies immediately, due to the complete breakdown of the membrane impermeability, or the initial membrane damage (i.e., increased membrane permeability) is repaired and the cell escapes immediate death. During both responses an increase of [Ca2+]in takes place in the target cells. In the cells that die immediately, however, [Ca2+]in reaches higher levels (approximately 1,400 nM) than in the cells that restore the initial damage (approximately 700 nM). Changes in target cell [pH]in are also detected during both responses. The direction of the change (acidification or alkalinization) as well as the level of the change depend on extracellular pH ([pH]ex). Also, [pH]in remains changed during the time the cells were followed (10 min). The programming time (i.e., the time from the initiation of the cytotoxic process to the time that a change in the physiological parameter was detected) of the killing process that leads to an immediate target cell death appears to be shortest at [pH]ex 7.3-7.6 (approximately 3 min).
    Original languageUndefined
    Pages (from-to)281-289
    Number of pages9
    JournalCytometry
    Volume20
    Issue number20
    DOIs
    Publication statusPublished - 1995

    Keywords

    • IR-60721
    • Calcium
    • Natural Killer cell
    • METIS-129719
    • fluorescent ratio probes
    • target cell
    • cytotoxic activity
    • pH

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