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

• Published in: Leukemia & lymphoma Vol. 23; no. 3-4; p. 313
• Main Authors: Yerly-Motta, V, Racadot, E, Fest, T, Bastard, C, Ferrand, C, Deschaseaux, M L, Herve, P
• Format: Journal Article
• Language: English
• Published: United States 1996
• Subjects: 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
• ISSN: 1042-8194
• DOI: 10.3109/10428199609054834

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.