title: Identification and structural analysis of rearranged immunoglobulin heavy chain genes in lymphomas and leukemias.
abstract: The third complementarity determining region (CDR3) of the hypervariable domain of immunoglobulin heavy chain (IgH) genes represents a highly variable and clone-specific IgH-CDR3 sequences in 10 non-Hodgkin's lymphomas (NHL), five chronic lymphocytic leukemias (CLL) and five acute lymphoblastic leukemias (ALL) of B cell lineage. The IgH-CDR3 sequences were amplified using DNA extracted from clinical specimens (bone marrow, peripheral blood and fresh-frozen or paraffin-embedded lymph nodes) by a semi-nested PCR with consensus primers directed to conserved regions within the variable (VH) and the joining (JH) gene segments. In 17/20 samples (85%), a distinct IgH-CDR3 PCR product was obtained. Individual PCR products were sequenced after cloning. The nucleotide sequences of 134 randomly chosen recombinant vectors were determined demonstrating in 17/20 cases (85%) monoclonal VH-N-DH-N-JH junctions. Analysis of PCR products by temperature-gradient gel electrophoresis (TGGE) confirmed the specificity of the IgH-CDR3 PCR/sequencing results. Moreover, the combination of PCR/TGGE technology allowed the rapid and specific characterization of clonal IgH-CDR3 junctions in B cell proliferations by direct sequencing even in the presence of admixed polyclonal B cells.
authors.affiliation: Department of Internal Medicine, Georg-August University, Goettingen, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Linke
authors.forename: J
authors.initials: J
authors.lastname: Pyttlich
authors.forename: M
authors.initials: M
authors.lastname: Tiemann
authors.forename: M
authors.initials: M
authors.lastname: Suttorp
authors.forename: R
authors.initials: R
authors.lastname: Parwaresch
authors.forename: W
authors.initials: W
authors.lastname: Hiddemann
authors.forename: M
authors.initials: M
authors.lastname: Kneba
mesh_terms: D001483:Base Sequence; D001706:Biopsy; D002051:Burkitt Lymphoma / Q000235:genetics*; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D004586:Electrophoresis / Q000379:methods; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain; D005803:Genes, Immunoglobulin; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000235:genetics*; D016393:Lymphoma, B-Cell / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014407:Tumor Cells, Cultured
chemical_list: D004273:DNA, Neoplasm; D007143:Immunoglobulin Heavy Chains