Immunoglobulin diversification in B cell malignancies: internal splicing of heavy chain variable region as a by-product of somatic hypermutation

Leukemia. 2002 Apr;16(4):636-44. doi: 10.1038/sj.leu.2402405.

Abstract

In this study we describe alternative splicing of somatically mutated immunoglobulin (Ig) variable heavy chain (V(H)) genes in three distinct primary B cell non-Hodgkin's lymphomas (B-NHL). In two V4-34 expressing lymphomas, ie a post-germinal center type B cell chronic lymphocytic leukemia (B-CLL) and a follicular lymphoma (FL), internally spliced V(H) gene transcripts were found in which a sequence stretch of 116 bp between the framework region 1 (FR1) and complementarity determining region 2 (CDR2) had been deleted. We provide evidence that for this alternative IgV(H) mRNA processing a known cryptic 5' splice donor site and a previously unidentified cryptic 3' splice acceptor site were used. Site-directed mutagenesis showed that the cryptic 3' splice acceptor site had been activated by specific somatic point mutations. The B-CLL further harbored a triplication of the rearranged JH3 gene segment including the putative N region and part of the JH3-JH4 intron sequence. This triplication probably took place via a repeated mechanism of DNA double strand break followed by homologous recombination, a mechanism which was recently proposed also involved in the somatic hypermutation process and is compatible with the post-germinal center derivation of this B-CLL. Finally, in a V4-34 expressing diffuse large B cell lymphoma, we observed alternative IgV(H) mRNA processing using the same cryptic 5' splice donor site and the normal splice acceptor site of the CH1-C(mu) exon. The significance of alternative IgV(H) processing in B cell malignancies and as a potential mechanism of somatic Ig diversification is discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Alternative Splicing / genetics*
  • Base Sequence
  • Complementarity Determining Regions / genetics
  • Consensus Sequence
  • DNA Primers / chemistry
  • DNA, Neoplasm / metabolism
  • Female
  • Genes, Immunoglobulin*
  • Humans
  • Immunoglobulin Heavy Chains / genetics*
  • Immunoglobulin Variable Region / genetics*
  • Leukemia, Lymphocytic, Chronic, B-Cell / genetics*
  • Leukemia, Lymphocytic, Chronic, B-Cell / metabolism
  • Leukemia, Lymphocytic, Chronic, B-Cell / pathology
  • Male
  • Molecular Sequence Data
  • Mutation*
  • Polymerase Chain Reaction
  • RNA, Neoplasm / metabolism
  • Sequence Homology, Nucleic Acid

Substances

  • Complementarity Determining Regions
  • DNA Primers
  • DNA, Neoplasm
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Variable Region
  • RNA, Neoplasm