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Chronic Lymphocytic Leukemia pp 51–62Cite as

Immunoglobulin Gene Analysis in Chronic Lymphocytic Leukemia

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1881))

Abstract

The formation of B-cell receptor immunoglobulin (BcR IG) is the result of a multi-step process that starts at the pro-B cell stage with the VDJ gene recombination of IG genes of the heavy chain, followed by VJ recombination of the light chain genes at the pre-B II cell stage. As a result, a fully functional BcR IG is expressed on the surface of any given naive B cell. After antigen encounter, somatic hypermutation (SHM) and class-switch recombination (CSR) act on the rearranged IG genes within the context of affinity maturation, leading to the expression of a BcR IG with unique immunogenetic and functional characteristics. Since B-cell neoplasms arise from the transformation of a single B cell, this renders IG gene rearrangements ideal clonal markers as they will be identical in all neoplastic cells of each individual clone. Furthermore, the rearranged IG sequence can also serve as a cell development/maturation marker, given that its configuration is tightly linked to specific B-cell developmental stages. Finally, in certain instances, as in the case of chronic lymphocytic leukemia (CLL), the clonotypic IG sequence and, more specifically, the load of somatic hypermutations within the rearranged IG heavy variable (IGHV) gene, holds prognostic and potentially predictive value. However, in order to take full advantage of the information provided from the analysis of the clonotypic IG gene rearrangement sequences, robust methods and tools need to be applied. Here, we provide details regarding the methodologies necessary to ensure reliable IG sequence analysis based on the recognized expertise of the European Research initiative on CLL (ERIC). All methodological and analytical steps are described below, starting from the isolation of blood mononuclear cells (PBMC), moving to the identification of the clonotypic IG rearrangement and ending with the accurate interpretation of the SHM status.

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Financial Support

Supported in part by H2020 “AEGLE, An analytics framework for integrated and personalized healthcare services in Europe,” by the EU; “MEDGENET, Medical Genomics and Epigenomics Network” (No.692298) by the EU; “TRANSCAN-179” NOVEL JTC 2016; “ESPA for young researchers 2017, Somatic hypermutation in Chronic Lymphocytic Leukemia: ontogenetic and clinical implications from the analysis of NGS immunogenetic data” (No. 5006864).

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Authors and Affiliations

  1. Institute of Applied Biosciences, Center for Research and Technology, Thessaloniki, Greece

    Andreas Agathangelidis, Anastasia Hadzidimitriou & Kostas Stamatopoulos

  2. Department of Molecular Medicine and Surgery, Karolinska Institute, Stockholm, Sweden

    Richard Rosenquist & Kostas Stamatopoulos

  3. Hôpital Pitié-Salpêtrière, Université Pierre et Marie Curie, Paris, France

    Frederic Davi

  4. IRCCS Istituto Scientifico San Raffaele, Università Vita-Salute San Raffaele, Milan, Italy

    Paolo Ghia

  5. Nikea G. Hospital, Athens, Greece

    Chrysoula Belessi

  6. Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden

    Kostas Stamatopoulos

Authors
  1. Andreas Agathangelidis
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  2. Richard Rosenquist
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  3. Frederic Davi
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  4. Paolo Ghia
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  5. Chrysoula Belessi
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  6. Anastasia Hadzidimitriou
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  7. Kostas Stamatopoulos
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Editor information

Editors and Affiliations

  1. Department of Internal Medicine, University of Michigan, Ann Arbor, MI, USA

    Sami N. Malek

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Agathangelidis, A. et al. (2019). Immunoglobulin Gene Analysis in Chronic Lymphocytic Leukemia. In: Malek, S. (eds) Chronic Lymphocytic Leukemia. Methods in Molecular Biology, vol 1881. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8876-1_5

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  • DOI: https://doi.org/10.1007/978-1-4939-8876-1_5

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8875-4

  • Online ISBN: 978-1-4939-8876-1

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