Intracytoplasmic detection of the Tac (p55) chain of interleukin-2 receptor in pre-B leukemic cells associated with a constitutive expression of Tac mRNA

Leukemia. 1990 Dec;4(12):819-25.

Abstract

The pre-B Reh-6 leukemic cells do not express membrane interleukin-2 (IL-2)-R alpha (Tac or p55) chain; however, their incubation with PMA induces the expression of both high and low affinity IL-2-R. Northern analysis of nonstimulated Reh-6 as well as leukemic cells from patients with acute B cell precursor lymphoblastic leukemia displayed a constitutive expression of p55 mRNA transcripts, which could be enhanced by PMA. Both actinomycin-D and cycloheximide could abrogate PMA-induced p55 membrane expression, suggesting the need for de novo mRNA and protein synthesis. The increased PMA-induced p55 mRNA accumulation was an early event (4 hr) and could be enhanced, specifically, by rIL-2 because anti-Tac moAb inhibited this rIL-2-mediated effect. Immunofluorescence and cross-linking studies using 125I-rIL-2 failed to reveal membrane-associated p55 protein on both Reh-6 and patients' leukemic cells. Conversely, immunogold staining and electron microscopy studies, revealed p55 immunoreactive molecules in the cytoplasm but not in the nucleus of all Reh-6 cells. Using a sensitive EIA, p55 molecules could be detected in cell lysates but not the culture supernatants of Reh-6 cells, suggesting that p55 was not released into the culture medium. These results indicate that constitutively expressed p55 mRNA on pre-B leukemic cells is translated into a relative immunoreactive protein that cannot be expressed on cell surface for unknown, yet, reasons.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Northern
  • Cell Division / drug effects
  • Cell Line / drug effects
  • Cell Membrane / metabolism
  • Cytoplasm / metabolism
  • Gene Expression / drug effects*
  • Humans
  • Membrane Proteins / metabolism
  • Microscopy, Electron
  • Precursor B-Cell Lymphoblastic Leukemia-Lymphoma / metabolism*
  • RNA, Messenger / metabolism*
  • Receptors, Interleukin-2 / genetics*
  • Receptors, Interleukin-2 / metabolism
  • Tetradecanoylphorbol Acetate / pharmacology

Substances

  • Membrane Proteins
  • RNA, Messenger
  • Receptors, Interleukin-2
  • Tetradecanoylphorbol Acetate