Flow cytometry to estimate circulating hematopoietic progenitors for autologous transplantation: comparative analysis of different CD34 monoclonal antibodies

Haematologica. 1991 Jul-Aug;76(4):330-3.

Abstract

We report that the 2 fluorescein (FITC) conjugated CD34 monoclonal antibodies available to date, namely FITC-8G12 and FITC-QBEND10, exhibit different capabilities of detecting circulating hematopoietic progenitors (CHP) as cells expressing the CD34 antigen (CD34+) by direct immunofluorescence flow cytometry. Mean fluorescence intensity conferred by FITC-QBEND10 to CD34+ CHP is 2.8-4.3 times lower than that conferred by FITC-8G12. By indirect immunofluorescence, native unconjugated QBEND10 and 8G12 antibodies detect CD34+ CHP in a comparable manner, thus indicating that the decrease in QBEND10 affinity is due to FITC-conjugation. Utilization of FITC-QBEND10 instead of FICT-8G12 to estimate infrequent (usually less than or equal to 5%) CD34+ CHP for clinical decision-making in autologous transplantation of CHP exposes clinicians to the risk of either overestimating (false positive) or underestimating (false negative) these cells in peripheral blood. Recently published guidelines for large-scale collection of CHP for autologous transplantation in cancer patients were based on data obtained with FITC-8G12. The same guidelines cannot be considered valid if FITC-QBEND10 is employed. We recommend FITC-8G12 as the optimal reagent to date for standardizing results of autologous CHP transplantation in cancer therapy.

Publication types

  • Comparative Study

MeSH terms

  • Antibodies, Monoclonal / immunology*
  • Antibody Specificity
  • Antigens, CD / analysis*
  • Antigens, CD34
  • Blood Cell Count / methods*
  • Blood Transfusion, Autologous
  • Breast Neoplasms / blood
  • Cell Separation
  • Flow Cytometry*
  • Fluorescein-5-isothiocyanate
  • Fluorescent Antibody Technique*
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Granulocyte-Macrophage Colony-Stimulating Factor / therapeutic use
  • Hematopoietic Stem Cell Transplantation
  • Hematopoietic Stem Cells* / drug effects
  • Hematopoietic Stem Cells* / immunology
  • Humans
  • Interleukin-3 / administration & dosage
  • Interleukin-3 / therapeutic use
  • Predictive Value of Tests
  • Recombinant Proteins / pharmacology
  • Recombinant Proteins / therapeutic use

Substances

  • Antibodies, Monoclonal
  • Antigens, CD
  • Antigens, CD34
  • Interleukin-3
  • Recombinant Proteins
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Fluorescein-5-isothiocyanate