Effect of anti-Ig on cytosolic Ca2+ in Daudi lymphoblastoid cells

D. A. Nachshen, Lawrence Pfeffer, I. Tamm

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Abstract

We examined the response in the free intracellular calcium concentration ([Ca2+](i)) of Daudi (human lymphoblastoid) cells to antibodies against human immunoglobulins (anti-Ig), and the relationship of [Ca2+](i) to anti-Ig-induced capping. At 80 μM intracellular quin-2 (a fluorescent probe for [Ca2+](i)), anti-Ig (10 μg/ml) caused a rapid increase in [Ca2+](i) from 100 to 600 nM; the signal returned to baseline with ~1 min. At 450 μM intracellular quin-2, [Ca2+](i) rose to only ~250 μM, and the signal declined gradually, returning to base line after >7 min. In subsequent experiments, the lower concentrations of quin-2 were employed. Plots of the amplitude of the [Ca2+](i) transients and of the binding of 125I-anti-Ig to Daudi cells versus the concentrations of anti-Ig showed similar saturation kinetics, with half-saturation occurring at 2-3 μg/ml. Part of the calcium in the transient is derived from the extracellular medium, and part from the nonmitochondrial intracellular stores. Caffeine (4 mM) and 8-(diethylamino)octyl 3,4,5-trimethoxybenzoate HCl (0.5 mM) suppressed the release of calcium from internal stores and the entry of calcium from outside the cells, but permitted capping in more than half of the cells. Phorbol esters (1-2 nM) inhibited both capping and the anti-Ig-induced decrease in [Ca2+](i). None of these agents blocked the binding of anti-Ig to the cells. It appears that receptor capping is not dependent on the anti-Ig-induced transient increase in calcium concentration.

Original languageEnglish (US)
Pages (from-to)15134-15139
Number of pages6
JournalJournal of Biological Chemistry
Volume261
Issue number32
StatePublished - Dec 1 1986

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Calcium
Receptor Aggregation
Phorbol Esters
Caffeine
Fluorescent Dyes
Immunoglobulins
Kinetics
Antibodies
Quin2
Experiments

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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Effect of anti-Ig on cytosolic Ca2+ in Daudi lymphoblastoid cells. / Nachshen, D. A.; Pfeffer, Lawrence; Tamm, I.

In: Journal of Biological Chemistry, Vol. 261, No. 32, 01.12.1986, p. 15134-15139.

Research output: Contribution to journalArticle

Nachshen, D. A. ; Pfeffer, Lawrence ; Tamm, I. / Effect of anti-Ig on cytosolic Ca2+ in Daudi lymphoblastoid cells. In: Journal of Biological Chemistry. 1986 ; Vol. 261, No. 32. pp. 15134-15139.
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