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

Menno C. Van Zelm, Magdalena A. Berkowska, Jacques J M Van Dongen

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

Abstract

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. The approach is useful to study basic B-cell biology as well as abnormal proliferation in human diseases.

Original languageEnglish
Title of host publicationLymphoma
Subtitle of host publicationMethods and Protocols
EditorsRalf Küppers
Place of PublicationNew York NY USA
PublisherHumana Press
Chapter6
Pages113-122
Number of pages10
ISBN (Electronic)9781627032698
ISBN (Print)9781627032681
DOIs
Publication statusPublished - 2013
Externally publishedYes

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
Volume971
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

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

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