Detection and monitoring of clonality in peripheral blood and bone marrow of patients with B‐cell lymphoproliferative disorders

Abstract

Bone marrow (BM) is accepted as the tissue of choice for the detection of monoclonal populations in leukemias and lymphomas; however, obtaining BM can be painful and traumatic for the patients. Although it is possible to detect clonality in peripheral blood (PB) samples, there are no reports comparing the results observed from BM with those from PB. Lymphoblastic leukemias and lymphomas are derived from B-lymphocytes in 80% of cases. In the early stages of their maturation, the immunoglobulin heavy chain genes (IgH) undergo rearrangements among their V, D, and J segments, giving rise to the Complementarity Determining Regions (CDR). Of these, CDR3 is unique for each lymphocyte and therefore it can be used as a tumour-specific marker in these malignant disorders. Among the 104 patients from whom we obtained pre-treatment paired samples of PB and BM, 94 (90.4%) showed concordant results. Similarly, at the end of treatment, 40 of 44 patients (90.9%) showed this concordance. During treatment only 24 patients were monitored and monoclones disappeared in 12 patients; in the other half, they persisted either partial or totally. We demonstrate that the detection and monitoring of monoclonal populations in the PB, in comparison with BM, was achieved with a statistical sensitivity of 90% and specificity of 92%.

Utility of Polymerase Chain Reaction for Analysis of Antigen Receptor Rearrangement in Staging and Predicting Prognosis in Dogs with Lymphoma

“…4 The limits of detection in dogs using currently available techniques is 1 neoplastic lymphocyte in a population of 100 heterogeneous, nonneoplastic lymphocytes. 4 In human patients, detection of clonality generally has been used to distinguish neoplasia from hyperplasia, 6,7 for detecting minimal residual disease after chemotherapy in leukemia, [8][9][10] and as a sensitive test for detecting circulating tumor cells in peripheral blood. 11 Sensitive detection of tumor cells has proven to have prognostic significance in some, but not all, situations.…”

High sensitivity of chemiluminescent methodology for detection of clonal CDR3 sequences in patients with acute lymphoblastic leukemia

“…The second (nested) PCR was performed with 5 0 and 3 0 primers of CDR3 consensus regions V and J respectively: CA1-25D, 5 0 CCG-AGG-ACA-CGG-CCG-TGT-ATT-ACT-G 3 0 and CA2-22D, 5 0 AAC-TGC-TGA-GGA-GAC-GGT-GAC-C 3 0 , 12 under the same thermal cycling conditions as the previous amplification, except that only 20 cycles of denaturation followed. 26 The expected nested PCR products ranged between 100 and 140 bp. PCR products were digested with the HaeIII and Sau96-1 restriction enzymes.…”

“…On the other hand, monitoring clonal behaviour during the induction treatment seems to be a complementary and useful procedure for establishing the prognosis of a patient with B-ALL. 2,3,26 …”