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Related Experiment Videos

Mitogen-activated Ca++ channels in human B lymphocytes

L H Brent1, Q Gong, J M Ross

  • 1Department of Medicine, Hahnemann University, Philadelphia, Pennsylvania 19102.

Journal of Cellular Physiology
|June 1, 1993
PubMed
Summary

Human B cell activation involves calcium influx through transmembrane potential-sensitive channels. These channels, regulated by inositol trisphosphate (IP3), are crucial for early B cell activation stages.

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Area of Science:

  • Immunology
  • Cell Biology
  • Biophysics

Background:

  • Human B cell activation is a complex process involving changes in intracellular calcium levels.
  • Mitogen-induced transmembrane conductances play a role in regulating calcium influx during B cell activation.

Purpose of the Study:

  • To investigate mitogen-induced transmembrane conductances in human B cells (Daudi cell line).
  • To elucidate the role of calcium influx and transmembrane potential in early B cell activation.

Main Methods:

  • Spectrofluorometry to measure intracellular calcium ([Ca++]i) and transmembrane potential.
  • Voltage clamp techniques to analyze membrane currents in Daudi cells.
  • Activation using anti-mu antibodies and inositol trisphosphate (IP3).

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Main Results:

  • Anti-mu antibody stimulation caused a biphasic rise in [Ca++]i, with the second phase due to extracellular calcium influx.
  • Calcium influx was sensitive to transmembrane potential and inhibited by high extracellular potassium.
  • Voltage clamp revealed inward currents activated by anti-mu antibodies and IP3, blocked by lanthanum, indicating Ca++ channel activity.
  • IP3 activation suggested a role for IP3 generation in conductance activation.

Conclusions:

  • Human B cells possess membrane calcium channels critical for early activation stages.
  • Transmembrane potential changes regulate calcium influx during B cell activation.
  • These mechanisms collectively control intracellular calcium levels in early B cell activation.