Studying the replication history of human B lymphocytes by real-time quantitative (RQ-)PCR

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The cells of the adaptive immune system, B and T lymphocytes, each generate a unique antigen receptor through V(D)J recombination of their immunoglobulin (Ig) and T-cell receptor (TCR) loci, respectively. Such rearrangements join coding elements to form a coding joint and delete the intervening DNA as circular excision products containing the signal joint. These excision circles are stable structures that cannot replicate and have no function in the cell. Since the coding joint in the genome is replicated with each cell division, the ratio between coding joints and signal joints in a population of B cells can be used as a measure for proliferation. This chapter describes a real-time quantitative (RQ-)PCR-based approach to quantify proliferation through calculating the ratio between coding joints and signal joints of the frequently occurring intronRSS–Kde rearrangements in the IGK light chain locus. Besides its use in normal B-cell biology, quantification of B-cell replication can inform on abnormal proliferation in human diseases and in B-cell neogenesis following depletion therapy.

Original languageEnglish
Title of host publicationLymphoma
Subtitle of host publicationMethods and Protocols
EditorsRalf Küppers
Place of PublicationNew York, NY
PublisherHumana Press
Number of pages12
ISBN (Electronic)9781493991518
ISBN (Print)9781493991501
Publication statusPublished - 1 Jan 2019

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • B cell
  • IGK
  • Immunoglobulin
  • IntronRSS
  • Kde
  • KREC
  • Proliferation
  • Real-time quantitative PCR
  • Replication history
  • V(D)J recombination

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