Comparative preclinical study of three bone marrow purging methods using PCR evaluation of residual t(14;18) lymphoma cells

Leuk Lymphoma. 1996 Oct;23(3-4):313-21. doi: 10.3109/10428199609054834.

Abstract

The t(14;18) chromosomal translocation occurring in most follicular lymphomas can be exploited by a Bcl2/JH polymerase chain reaction (PCR) to detect residual disease and to monitor the effectiveness of ex-vivo tumor cell immunological purging. We first demonstrated the 10(-5) Bcl2/JH PCR sensitivity with serial dilutions of OCY-LY8 lymphoma cell lines in normal mononuclear cells; and then the specificity and reproductibility of this technique by analysing follicular and non follicular lymphoma samples. With the Bcl2/JH PCR, we tested the efficiency of three marrow purging protocols with an experimentally contaminated bone marrow either treated by three anti-B cell monoclonal antibodies (mAb) followed by three rounds of rabbit complement or two rounds of immunomagnetics beads. Samples obtained after each purging were amplified by Bcl2/JH PCR and hybridized with PFL3 probe. We were able to produce a 2 to 3 log tumor cell reduction after three rounds of complement and a 4 to 5 log reduction after two rounds of beads. This study showed that it is feasible to use the Bcl2/JH PCR technique for residual cell lymphoma detection in patients undergoing intensive chemotherapy or BM transplantation. These results indicate that ex-vivo immunomagnetic BM purging is probably superior to complement mediated lysis for the eradication of B lymphoma cells from the marrow of patients undergoing autologous transplantation.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Bone Marrow Purging / methods*
  • Chromosomes, Human, Pair 14*
  • Chromosomes, Human, Pair 18*
  • Flow Cytometry
  • Humans
  • Immunomagnetic Separation
  • Lymphoma, Follicular / diagnosis
  • Lymphoma, Follicular / genetics*
  • Lymphoma, Follicular / pathology
  • Neoplasm, Residual
  • Polymerase Chain Reaction / methods*
  • Rabbits
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Translocation, Genetic*