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

Menno C. van Zelm*

*Corresponding author for this work

Research output: Chapter/Conference proceedingChapterAcademic

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. 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 publicationMethods in Molecular Biology
Pages127-138
Number of pages12
DOIs
Publication statusPublished - 2019
Externally publishedYes

Publication series

SeriesMethods in Molecular Biology
Volume1956
ISSN1064-3745

Bibliographical note

Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2019.

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