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Studies on leukemic cell tissue factor.

H Tanaka1, N Narahara, H Kurabayashi

  • 1Third Department of Internal Medicine, Gunma University School of Medicine, Japan.

Thrombosis Research
|March 15, 1989
PubMed
Summary
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Researchers purified human placental apoprotein and analyzed tissue factor in leukemia cells. Endotoxin stimulation increased procoagulant activity and tissue factor expression on leukemia cell surfaces.

Area of Science:

  • Biochemistry
  • Hematology
  • Cell Biology

Background:

  • Tissue factor is a key initiator of the coagulation cascade.
  • Leukemia cells can express tissue factor, potentially contributing to thrombotic complications.

Purpose of the Study:

  • To purify the apoprotein component of human placental tissue factor.
  • To investigate the presence and expression of tissue factor in human leukemia cells.
  • To analyze the cellular localization and regulation of tissue factor in response to endotoxin stimulation.

Main Methods:

  • Concanavalin A-Sepharose affinity chromatography and SDS-PAGE for protein purification.
  • Generation of specific rabbit anti-tissue factor IgG for immunological analysis.
  • Immunostaining and immuno-electron microscopy to detect and localize tissue factor.

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Main Results:

  • Human placental apoprotein was purified with a molecular weight of 45,000 (non-reduced) and 49,000 (reduced).
  • Tissue factor was detected in human leukemia cells (M2, M3) and cell lines (HL-60, Molt-4).
  • Endotoxin stimulation of HL-60 and Molt-4 cells enhanced procoagulant activity and increased tissue factor localization on cell surface pseudopods and endoplasmic reticulum.

Conclusions:

  • Purified human placental apoprotein serves as a reliable antigen for antibody production.
  • Leukemia cells express tissue factor, and its surface expression is upregulated by endotoxin.
  • This upregulation involves increased synthesis and translocation to the cell surface, particularly on membrane protrusions.