Characterization of the immunoglobulin heavy chain complementarity determining region (CDR)-III sequences from human B cell precursor acute lymphoblastic leukemia cells.

Abstract

Sequence analysis of the immunoglobulin heavy chain complementarity determining region (CDR)-III of B-lineage cells at various stages has provided important insights concerning B cell maturation and selection. Knowledge of human CDR-III sequences has been relatively limited compared with that of the murine system. We analyzed the CDR-III sequences of B cell precursor acute lymphoblastic leukemia (pre-B ALL) cells in 23 newly diagnosed and 10 relapsed patients, in order to elucidate the organization of CDR-III in B cell precursors. We found a very low frequency of somatic mutations in D and JH regions, preferential use of DLR, Dxp, DHQS2, and DN elements, and of 3' side JH segments, and no predominant usage of D coding frames. Unusual joinings such as VH-D-D-JH and VH_ JH were observed in three, and one sequences, respectively. We compared the CDR-III sequences derived from 10 patients between diagnosis and relapse. Two of them had three spots of mutated nucleotides at relapse, all of which were found in the N region near the D segments. Our data showed the possibility of somatic mutation at relapse, in addition to developmentally regulated rearrangement of the immunoglobulin gene at the stage of B cell precursors. (J. Clin. Invest. 1992. 89:739-746.)

Comparison of the immunoglobulin heavy‐chain complementarity determining region‐3 structure among the DNA sequences and the μ‐ and γ‐transcripts in human B‐lineage cells

“…Forty cycles of PCR were performed as described. 6,13 Amplified products were separated through 8% polyacrylamide gels, and slices spanning the 70-200 bp region were isolated. The fragments were digested with SalI and HindIII restriction endonucleases (Boehringer Mannheim Yamanouchi, Tokyo, Japan), then subcloned into the M13 mp18/19 phage vector (Boehringer Mannheim Yamanouchi).…”

“…6,15,16 DNA was sequenced using the fluoresceinconjugated M13 universal primer on a DNA sequencer (373-A; Applied Biosystems, Inc., Foster City, CA) according to the manufacturer's instructions. Cross contamination was prevented throughout this study as described.…”

Utilization of Ig heavy chain variable, diversity, and joining gene segments in children with B-lineage acute lymphoblastic leukemia: implications for the mechanisms of VDJ recombination and for pathogenesis

“…4 The CDR3 sequence is unique to each rearrangement and therefore identifies individual B cells or clonal B-cell expansion. 4,14,15 The V H D H J H gene repertoires are restricted and developmentally regulated at early stages of differentiation. 3,16 In the most immature B-cell precursors (pro-B cells), the IgH genes remain germ line or there is only D H J H joining.…”

Characterization of the immunoglobulin light chain variable region gene expressed in multiple myeloma

“…In the rearrangement of the IgH gene, the usage of J H segments revealed a developmental stage-specific trend. [7][8][9][10][11][12][13] At the earliest stage of B cell development, the initial D-J H rearrangements predominantly occur between D H Q52 and J H -1 segments which are located close to each other. 53 During B cell development, a second or a third D-J H joining might occur, then the more 5ЈD segments and more 3ЈJ H segments would join at a later stage.…”

“…6 Studies on IgH variable region gene sequences at various stages of development have revealed stage-specific trends in the usage of V H , D, and J H segments, the degree of N nucleotide addition, and the rate of somatic mutations. [7][8][9][10][11][12][13] However, little is known of the organization and diversification of the IgL variable region gene on -chain. On the other hand, multiple myeloma (MM) has a variety of clinical features relating to clinical stage, prognosis and complication.…”