scholarly journals VH gene rearrangement events can modify the immunoglobulin heavy chain during progression of B-lineage acute lymphoblastic leukemia [see comments]

Blood ◽  
1992 ◽  
Vol 79 (1) ◽  
pp. 223-228 ◽  
Author(s):  
R Wasserman ◽  
M Yamada ◽  
Y Ito ◽  
LR Finger ◽  
BA Reichard ◽  
...  
Keyword(s):  
Heavy Chain ◽  
Gene Rearrangement ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Immunoglobulin Heavy Chain ◽  
Gene Replacement ◽  
Vh Gene ◽  
Polymerase Chain ◽  
B Lineage ◽  
Complementarity Determining Region

The presence of multiple VHDJH joinings in upwards of 30% of acute lymphoblastic leukemias (ALL) suggests a relative instability of the rearranged immunoglobulin heavy chain (IgH) gene, but the mechanisms involved are not completely understood. An investigation of the structure of the VHDJH joinings using complementarity determining region (CDR)3 polymerase chain reaction (PCR) in 12 leukemias at both diagnosis and relapse indicates that this instability may increase as a function of time. In only one of seven cases in which relapse occurred within 3 years from diagnosis was a new VHDJH joining identified and this coexisted with the original diagnostic joining. Most strikingly, new VHDJH joinings were identified in four of five cases in which relapse occurred more than 5 years from diagnosis. In this latter population, the instability of the joinings was generated from VH----VH gene replacement events in two cases, since the new joinings retained the original DJH sequences and partial N region homology at the VHD junction, and probably in a third case from a VH gene rearrangement to a common DJH precursor. Furthermore, in five of 23 (21.7%) additional cases studied at diagnosis, subclones were identified that had similar modifications of the VH-N region. These data indicate that VH gene replacement events and VH gene rearrangements to a common DJH joining contribute to the instability of the VHDJH joining in ALL. This phenomenon should be taken into consideration in those methodologies that exploit IgH rearrangements for detection of minimal residual disease.

VH gene rearrangement events can modify the immunoglobulin heavy chain during progression of B-lineage acute lymphoblastic leukemia [see comments]

Blood ◽  
1992 ◽  
Vol 79 (1) ◽  
pp. 223-228 ◽  
Author(s):  
R Wasserman ◽  
M Yamada ◽  
Y Ito ◽  
LR Finger ◽  
BA Reichard ◽  
...  
Keyword(s):  
Heavy Chain ◽  
Gene Rearrangement ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Immunoglobulin Heavy Chain ◽  
Gene Replacement ◽  
Vh Gene ◽  
Polymerase Chain ◽  
B Lineage ◽  
Complementarity Determining Region

Abstract The presence of multiple VHDJH joinings in upwards of 30% of acute lymphoblastic leukemias (ALL) suggests a relative instability of the rearranged immunoglobulin heavy chain (IgH) gene, but the mechanisms involved are not completely understood. An investigation of the structure of the VHDJH joinings using complementarity determining region (CDR)3 polymerase chain reaction (PCR) in 12 leukemias at both diagnosis and relapse indicates that this instability may increase as a function of time. In only one of seven cases in which relapse occurred within 3 years from diagnosis was a new VHDJH joining identified and this coexisted with the original diagnostic joining. Most strikingly, new VHDJH joinings were identified in four of five cases in which relapse occurred more than 5 years from diagnosis. In this latter population, the instability of the joinings was generated from VH----VH gene replacement events in two cases, since the new joinings retained the original DJH sequences and partial N region homology at the VHD junction, and probably in a third case from a VH gene rearrangement to a common DJH precursor. Furthermore, in five of 23 (21.7%) additional cases studied at diagnosis, subclones were identified that had similar modifications of the VH-N region. These data indicate that VH gene replacement events and VH gene rearrangements to a common DJH joining contribute to the instability of the VHDJH joining in ALL. This phenomenon should be taken into consideration in those methodologies that exploit IgH rearrangements for detection of minimal residual disease.

scholarly journals Prospective monitoring and quantitation of residual blasts in childhood acute lymphoblastic leukemia by polymerase chain reaction study of delta and gamma T-cell receptor genes

Blood ◽  
1994 ◽  
Vol 83 (7) ◽  
pp. 1892-1902 ◽  
Author(s):  
H Cave ◽  
C Guidal ◽  
P Rohrlich ◽  
MH Delfau ◽  
A Broyart ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
T Cell Receptor ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Cell Receptor ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
B Lineage ◽  
Minimal Residual

Abstract We have developed a strategy based on polymerase chain reaction (PCR) for detecting all possible gamma T-cell receptor (gamma TCR) rearrangements and the most common delta TCR rearrangements found in B- lineage and T-acute lymphoblastic leukemia (T-ALL). The segments amplified from blasts are then directly sequenced to derive clonospecific probes. From a series of 45 patients aged 1 to 15 years (42 B-lineage ALL, 3 T-ALL), 35 (83%) could be followed for minimal residual disease with at least one clonospecific probe. Detection of clonal markers using clonospecific probes routinely allowed the detection of 1 to 10 blasts out of 10(5) cells as determined by serial dilutions of the initial samples. Residual disease was quantitated by a competitive PCR assay based on the coamplification of an internal standard. Twenty children were prospectively followed for periods varying from 7 to 30 months. In most children, a progressive decrease of the tumor load was observed, and blasts became undetectable within 6 months after the initiation of treatment. A slower kinetics of decrease in tumor cells was found in three children. These three patients relapsed with blasts that continued to display the initial clonospecific markers. Three other children had a central nervous system relapse despite the absence of detectable medullary residual disease. The use of both delta and gamma TCR genes as clonal markers, as well as simplification in the methods to detect and quantify residual blasts reported here, will allow the study of the large number of patients required to determine the role of the detection of minimal residual disease by PCR in the follow-up of childhood ALL.

scholarly journals Clonal evolution in B-lineage acute lymphoblastic leukemia by contemporaneous VH-VH gene replacements and VH-DJH gene rearrangements

Blood ◽  
1996 ◽  
Vol 87 (6) ◽  
pp. 2506-2512 ◽  
Author(s):  
Y Choi ◽  
SJ Greenberg ◽  
TL Du ◽  
PM Ward ◽  
PM Overturf ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
B Cell ◽  
Lymphoblastic Leukemia ◽  
Clonal Evolution ◽  
Gene Segment ◽  
Gene Replacement ◽  
Gene Rearrangements ◽  
Region Gene ◽  
Vh Gene ◽  
B Lineage

B-cell acute lymphoblastic leukemia (B-ALL), more frequently than any other B-lineage neoplasm, exhibits oligoclonal Ig heavy chain (IgH) gene rearrangement in 15% to 43% of all cases studied. To study the molecular processes that promote multiple IgH rearrangements, a comprehensive sequence analysis of a B-ALL case was performed in which seven clonal IgH gene rearrangements were identified. The genetic profiles suggested that a single leukemic progenitor clone evolved into several subclones through dual processes of variable (VH) to preexisting diversity-joining (DJH) gene segment rearrangement and VH to VH gene replacement. Predominant IgH-V usage and the uniquely rearranged clonotype-specific VHDJH region gene sequences were identified using a novel DNA-based gene amplification strategy. Polymerase chain reaction (PCR) was directed by an IgH-J generic primer and a complement of family-specific IgH-V primers that defined the major B-cell IgH-V gene usage. Clonality of rearranged VHDJH bands was substantiated by high resolution denaturant gel electrophoretic analysis. Sequence patterns of the amplified VHDJH fragments segregated into two groups defined by common DJH sequences. Partial N region homology at the VHD junction as well as shared DJH sequences firmly established VH to VHDJH gene replacement as a mechanism generating clonal evolution in one group. In the second subset, oligoclonality was propagated by independent VH gene rearrangements to a common DJH precursor. The contributions of all clonal Ig-VHDJH repertoires for each group was approximately 50% and reflected a symmetric distribution of leukemic subclones generated by either process. Thus, oligoclonal rearrangements evolved by two independent, yet seemingly contemporaneous molecular genetic mechanisms. All seven clones displayed nonfunctional Ig-VHDJH recombinations. These observations may have relevance to the recombinatorial opportunities available during normal B-cell maturation.

scholarly journals Prospective monitoring and quantitation of residual blasts in childhood acute lymphoblastic leukemia by polymerase chain reaction study of delta and gamma T-cell receptor genes

Blood ◽  
1994 ◽  
Vol 83 (7) ◽  
pp. 1892-1902 ◽  
Author(s):  
H Cave ◽  
C Guidal ◽  
P Rohrlich ◽  
MH Delfau ◽  
A Broyart ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
T Cell Receptor ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Cell Receptor ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
B Lineage ◽  
Minimal Residual

We have developed a strategy based on polymerase chain reaction (PCR) for detecting all possible gamma T-cell receptor (gamma TCR) rearrangements and the most common delta TCR rearrangements found in B- lineage and T-acute lymphoblastic leukemia (T-ALL). The segments amplified from blasts are then directly sequenced to derive clonospecific probes. From a series of 45 patients aged 1 to 15 years (42 B-lineage ALL, 3 T-ALL), 35 (83%) could be followed for minimal residual disease with at least one clonospecific probe. Detection of clonal markers using clonospecific probes routinely allowed the detection of 1 to 10 blasts out of 10(5) cells as determined by serial dilutions of the initial samples. Residual disease was quantitated by a competitive PCR assay based on the coamplification of an internal standard. Twenty children were prospectively followed for periods varying from 7 to 30 months. In most children, a progressive decrease of the tumor load was observed, and blasts became undetectable within 6 months after the initiation of treatment. A slower kinetics of decrease in tumor cells was found in three children. These three patients relapsed with blasts that continued to display the initial clonospecific markers. Three other children had a central nervous system relapse despite the absence of detectable medullary residual disease. The use of both delta and gamma TCR genes as clonal markers, as well as simplification in the methods to detect and quantify residual blasts reported here, will allow the study of the large number of patients required to determine the role of the detection of minimal residual disease by PCR in the follow-up of childhood ALL.

scholarly journals Massive evolution of the immunoglobulin heavy chain locus in children with B precursor acute lymphoblastic leukemia

Blood ◽  
2012 ◽  
Vol 120 (22) ◽  
pp. 4407-4417 ◽  
Author(s):  
Charles Gawad ◽  
Francois Pepin ◽  
Victoria E. H. Carlton ◽  
Mark Klinger ◽  
Aaron C. Logan ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
B Cells ◽  
B Cell ◽  
Heavy Chain ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Clonal Evolution ◽  
Immunoglobulin Heavy Chain ◽  
Igh Locus ◽  
Precursor B Cells

Abstract The ability to distinguish clonal B-cell populations based on the sequence of their rearranged immunoglobulin heavy chain (IgH) locus is an important tool for diagnosing B-cell neoplasms and monitoring treatment response. Leukemic precursor B cells may continue to undergo recombination of the IgH gene after malignant transformation; however, the magnitude of evolution at the IgH locus is currently unknown. We used next-generation sequencing to characterize the repertoire of IgH sequences in diagnostic samples of 51 children with B precursor acute lymphoblastic leukemia (B-ALL). We identified clonal IgH rearrangements in 43 of 51 (84%) cases and found that the number of evolved IgH sequences per patient ranged dramatically from 0 to 4024. We demonstrate that the evolved IgH sequences are not the result of amplification artifacts and are unique to leukemic precursor B cells. In addition, the evolution often follows an allelic exclusion pattern, where only 1 of 2 rearranged IgH loci exhibit ongoing recombination. Thus, precursor B-cell leukemias maintain evolution at the IgH locus at levels that were previously underappreciated. This finding sheds light on the mechanisms associated with leukemic clonal evolution and may fundamentally change approaches for monitoring minimal residual disease burden.

scholarly journals Minimal Residual Disease Evaluation By Real-Time Quantitative Polymerase Chain Reaction for Wilms Tumor 1 Gene Expression and Multicolour Flow Cytometry, in Pediatric B Lineage Acute Lymphoblastic Leukemia

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 4985-4985
Author(s):  
Neelam Varma ◽  
Sandeep R. ◽  
Shano Naseem ◽  
Man Updesh Singh Sachdeva ◽  
Deepak Bansal ◽  
...  
Keyword(s):  
Gene Expression ◽  
Flow Cytometry ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Wt1 Gene ◽  
Chain Reaction ◽  
Over Expression ◽  
Polymerase Chain ◽  
B Lineage ◽  
Minimal Residual

Abstract Introduction: Minimal residual disease (MRD) determination involves the measurement of very low levels of leukemia using sensitive techniques which at present are complex, time consuming and require expertise for performance and interpretation. A ''panleukemic''marker such as Wilm's tumor 1 gene (WT1) which is frequently over expressed in acute leukemia could simplify MRD detection and serve as a useful prognostic marker. Aim of study: To evaluate the usefulness of WT1 gene expression, as a marker for MRD in B-lineage acute lymphoblastic leukemia (B-ALL). Method and Material: Flow cytometric immunophenotyping (FCMI) and real time-polymerase chain reaction (RQ-PCR) for WT1 gene expression were performed usingbone marrow at diagnosis and at day 15 (mid-induction). Of the 23 patients recruited, day 15 MRD analyses by both these methods was performed on 11 bone marrow samples of patients who showed WT1 over expression at day 0. Results: WT1 over expression at diagnosis was found in 69.5% cases (16/23). MRD was detectable in 54.5% cases by WT1 RQ-PCR and by FCM in 72% cases. A statistically significant correlation was seen between WT1 normalized copy number (NCN) at diagnosis with MRD levels detected by FCM. Conclusion: WT1 represents a candidate MRD and prognostic marker. The significant correlation between WT1 over expression at diagnosis and MRD positivity by flow cytometry at day 15 (mid induction) of chemotherapy suggests that high WT1 expression could correlate with unfavourable outcome in childhood ALL. However, as not all patients of B-ALL over-express WT1 at diagnosis, quantitative assessment of WT1 transcripts can be used as useful molecular marker for MRD detection, but only in a subset of patients. Disclosures No relevant conflicts of interest to declare.

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