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The Journal of Immunology, Vol 150, Issue 7 2620-2633, Copyright © 1993 by American Association of Immunologists
ARTICLES |
SD Hess, M Oortgiesen and MD Cahalan
Department of Physiology and Biophysics, University of California, Irvine 92717.
With the use of single-cell digital imaging of the fluorescent Ca2+ indicator dye fura-2 we investigated Ca2+ signaling in human T lymphocytes and NK cells during activation by a variety of stimuli. A low percentage of resting T cells or T cell blasts displayed oscillations in cytosolic Ca2+ when stimulated with the mitogenic lectin PHA or by the addition of OKT3 mAb followed by a secondary cross- linking antibody. Lymphokine-activated T killer cells were more responsive than resting cells. A comparison of PHA, cross-linked anti- CD3, and a heteroconjugate mAb showed that at least 20% of the cells from these T cell preparations oscillated. Addition of PHA or cross- linked anti-CD16 caused NK cells to oscillate. In contrast, thapsigargin, a microsomal ATPase blocker, resulted in a relatively uniform, slowly rising and sustained Ca2+ response in all cell types studied. The maintenance of both thapsigargin- and receptor-induced responses required Ca2+ influx driven by a negative membrane potential. Because Ca2+ oscillations occurred in response to stimuli which mimic the normal activation of lymphocytes, and inasmuch as the percentage of oscillating cells increases with state of activation, these oscillations may play an important role in mitogenic activation.
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