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Subcellular Fractionation of Primary Chronic Lymphocytic Leukemia Cells to Monitor Nuclear/Cytoplasmic Protein Trafficking
11:39

Subcellular Fractionation of Primary Chronic Lymphocytic Leukemia Cells to Monitor Nuclear/Cytoplasmic Protein Trafficking

Published on: October 23, 2019

PRAME is a membrane and cytoplasmic protein aberrantly expressed in chronic lymphocytic leukemia and mantle cell

Rodrigo Proto-Siqueira1, Lorena L Figueiredo-Pontes, Rodrigo A Panepucci

  • 1Hematology Division and Center for Cell Based Therapy, Medical School of Ribeirão Preto, University of São Paulo, Brazil.

Leukemia Research
|April 20, 2006
PubMed

Insights

Preferentially expressed antigen in melanoma (PRAME) is highly expressed in chronic lymphoproliferative disorders (CLD) but not in healthy individuals. PRAME protein analysis can help distinguish leukemic cells in CLD and may be a therapeutic target.

Area of Science:

  • Hematology
  • Immunology
  • Oncology

Background:

  • Aberrant expression of the preferentially expressed antigen in melanoma (PRAME) gene is observed in chronic lymphoproliferative disorders (CLD).
  • PRAME's role in distinguishing malignant from normal cells in CLD requires further investigation.

Purpose of the Study:

  • To develop and validate a quantitative flow cytometric (QFC) method for evaluating PRAME protein expression in CLD.
  • To assess the diagnostic utility of PRAME expression for differentiating CLD from normal B lymphocytes and peripheral blood mononuclear cells (PBMCs).
  • To explore PRAME as a potential therapeutic target in CLD.

Main Methods:

  • Production and characterization of a novel anti-PRAME monoclonal antibody (MoAb).
  • Application of a quantitative flow cytometric (QFC) method to measure PRAME expression in peripheral blood (PB) of 47 CLD patients and 15 healthy subjects.
  • Immunofluorescence microscopy and permeabilized flow cytometry to determine PRAME localization.

Main Results:

  • Approximately 90% of CLD patients exhibited >20% PRAME+ lymphocytes, while normal samples showed no significant expression.
  • CLD cells displayed significantly higher PRAME expression intensity compared to normal B lymphocytes and PBMCs.
  • PRAME was confirmed as both a membrane antigen and a cytoplasmic protein aberrantly expressed in malignant CLD cells.

Conclusions:

  • Quantitative analysis of PRAME protein expression is a valuable tool for distinguishing leukemic cells in CLD from normal cells.
  • Aberrant PRAME expression in CLD suggests its potential as a diagnostic biomarker.
  • PRAME represents a promising molecular target for novel therapeutic strategies in CLD.

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