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Detection of minimal residual disease in B chronic lymphocytic leukemia (CLL)
1Unité d'Immuno-Hématologie et d'Immunopathologie, Institut Pasteur, Paris, France.
Insights
A sensitive method for detecting minimal residual disease (MRD) in chronic lymphocytic leukemia (CLL) patients was developed. This approach, utilizing immunoglobulin gene rearrangement, achieved molecular remission (MR) in some patients, particularly those undergoing autologous bone marrow transplantation.
Area of Science:
- Hematology
- Oncology
- Molecular Biology
Background:
- Detecting minimal residual disease (MRD) is crucial for managing B cell malignancies like chronic lymphocytic leukemia (CLL).
- Immunoglobulin (Ig) gene rearrangement offers a unique marker for B cell clones in the absence of specific chromosomal translocations.
- Assessing MRD in patients achieving complete hematological remission (CHR) is essential for monitoring treatment efficacy.
Purpose of the Study:
- To develop and report a highly sensitive method for assessing minimal residual disease (MRD) in chronic lymphocytic leukemia (CLL) patients.
- To evaluate the method's ability to detect MRD in patients in complete hematological remission (CHR) and complete phenotypic remission (CPR).
- To investigate the achievement of molecular remission (MR) in CLL patients treated with different therapeutic regimens.
Main Methods:
- Inclusion of twelve CLL patients in CHR and CPR, treated with Fludarabine (FDR), CHOP regimen, or autologous bone marrow transplantation (ABMT).
- DNA extraction from peripheral blood lymphocytes (PBL) followed by polymerase chain reaction (PCR) amplification using VH family specific and JH primers.
- Utilized CDR3 sequence specific probes as 3' primers when initial PCR assays were negative, with PCR products analyzed by gel electrophoresis.
Main Results:
- The method demonstrated high sensitivity, detecting a single leukemic cell among 10(5) normal cells.
- Four out of 12 patients achieved molecular remission (MR) using CDR3 amplification; all 3 autografted patients achieved MR.
- Clone-specific probes confirmed clonal signals in most cases, highlighting the potential for achieving MR in B-CLL.
Conclusions:
- The developed method is highly sensitive for assessing MRD in CLL patients in remission.
- Molecular remission (MR) can be achieved in a subset of B-CLL patients, particularly after autologous bone marrow transplantation.
- Further research is necessary to establish the precise correlation between MRD levels and clinical outcomes in CLL.
Unlabelled:
In the absence of specific chromosomal translocations the best method for detecting minimal residual disease (MRD) in B cell malignancies is based on the uniqueness of immunoglobulin (Ig) genes rearrangement. We here report a very sensitive method for assessing MRD in complete hematological remission (CHR) chronic lymphocytic leukemia (CLL) patients as defined by the international workshop on CLL (IWCLL).
Patients:
Twelve CLL patients in CHR and complete phenotypic remission (CPR) were included in the study. Eight of them received Fludarabine (FDR), one was treated by Chop regimen, and the remaining 3 were rescued by polychemotherapy followed by autologous bone marrow transplantation (ABMT).
Methods:
DNA extracted from peripheral blood lymphocytes (PBL) of each patient was amplified with VH family specific and framework 3 primers in 5' and a consensus JH primer in 3', before treatment and sequentially after the CPR completion. When no clonal rearrangement could be detected by this assay, the CDR3 sequence specific probe of the clone was used as the 3' primer, associated to the VH family specific primer in 5'. PCR products were analyzed by classical procedures in agarose and/or acrylamide gels.
Results:
Mixtures of leukemic cells and normal PBL showed detection of a single leukemic cell among more than 10(5) normal cells. Four out of the 12 patients achieved molecular remission (MR) when employing CDR3 amplification. All 3 autografted patients were in MR, whereas only one out of the 9 patients treated by chemotherapy alone achieved MR. When using a clone specific probe, a clonal signal was observed in all cases but one (ABMT). Results presented here confirm that MR may be achieved in a few cases of B-CLL. Further studies are needed to determine the exact relationship between MRD and clinical outcome.
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