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Real-Time Quantitative (RQ-)PCR Approach to Quantify the Contribution of Proliferation to B Lymphocyte Homeostasis

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Immune Homeostasis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 979))

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 relatively 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 polymerase chain reaction (RQ-PCR)-based approach to quantify proliferation through calculating the ratio between coding joints and signal joints of the frequently occurring kappa-deleting rearrangements in the IGK light chain loci in man and mouse. The approach is useful to study the contribution of proliferation to B-cell homeostasis in health and disease.

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Acknowledgments

MAB is supported by a Fellowship from the Ter Meulen Fund–Royal Netherlands Academy of Arts and Sciences. MCvZ is supported by an Erasmus University Rotterdam (EUR)-Fellowship, an Erasmus MC Fellowship, by Veni grant 916.110.90 from ZonMW/NWO and by grant 689 of the Sophia Children’s Hospital Fund.

Conflict of interest

JJMvD is inventor of the KREC assay, which has been patented (PCT/NL 2005/000761; priority date 25 Oct 2004) and licensed to InVivoScribe Technologies, San Diego, CA; revenues of the patent go to Erasmus MC. The other authors have declared that no conflict of interest exists.

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Authors and Affiliations

  1. Department of Immunology, Erasmus MC, University Medical Center, Rotterdam, The Netherlands

    Menno C. van Zelm, Magdalena A. Berkowska, Mirjam van der Burg & Jacques J. M. van Dongen

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  1. Menno C. van Zelm
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  2. Magdalena A. Berkowska
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  3. Mirjam van der Burg
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  4. Jacques J. M. van Dongen
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Corresponding author

Correspondence to Menno C. van Zelm .

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Editors and Affiliations

  1. of the Health Sciences, Department of Pharmacology, Uniformed Services University, Jones Bridge Road 4301, Bethesda, 20814, Maryland, USA

    Andrew L. Snow

  2. NIAID, Laboratory of Immunology, National Institutes of Health, Rockville Pike 9000, Bethesda, 20892, Maryland, USA

    Michael J. Lenardo

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van Zelm, M.C., Berkowska, M.A., van der Burg, M., van Dongen, J.J.M. (2013). Real-Time Quantitative (RQ-)PCR Approach to Quantify the Contribution of Proliferation to B Lymphocyte Homeostasis. In: Snow, A., Lenardo, M. (eds) Immune Homeostasis. Methods in Molecular Biology, vol 979. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-290-2_11

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  • DOI: https://doi.org/10.1007/978-1-62703-290-2_11

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-289-6

  • Online ISBN: 978-1-62703-290-2

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