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Determining the Origin of Human Germinal Center B Cell-Derived Malignancies

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Germinal Centers

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1623))

Abstract

Most human B cell lymphomas originate from germinal center (GC) B cells. This is partly caused by the high proliferative activity of GC B cells and the remodeling processes acting at the immunoglobulin (Ig) loci of these cells, i.e., somatic hypermutation and class-switching. Mistargeting of these processes can cause chromosomal translocations, and the hypermutation machinery may also target non-Ig genes. As somatic hypermutation is exclusively active in GC B cells, the presence of somatic mutations in rearranged IgV genes is a standard criterium for a GC or post-GC B cell origin of lymphomas. Beyond this, ongoing somatic hypermutation during lymphoma clone expansion indicates that the lymphoma has an active GC B cell differentiation program. The proto-oncogene BCL6 is specifically expressed in GC B cells and also acquires somatic mutations as a physiological by-product of the somatic hypermutation process, albeit at a lower level than IgV genes. Thus, detection of BCL6 mutations is a further genetic trait of a GC experience of a B cell lymphoma. Typically, B cell lymphomas retain key features of their specific cells of origin, including a differentiation stage-specific gene expression pattern. This is at least partly due to genetic lesions, which “freeze” the lymphoma cells at the differentiation stage at which the transformation occurred. Therefore, identification of the normal B cell subset with the most similar gene expression pattern to a particular type of B cell lymphoma has been instrumental to deduce the precise cell of origin of lymphomas.

We present here protocols to analyze human B cell lymphomas for a potential origin from GC B cells by determining the presence of mutations in rearranged IgV genes and the BCL6 gene, and by comparing the gene expression pattern of lymphoma cells with those of normal B cell subsets by genechip or RNA-sequencing analysis.

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Acknowledgments

We thank Kerstin Heise and Ludger Klein-Hitpass for help in gathering the protocols. Our own work on lymphoma pathogenesis and GEP is supported by the Deutsche Forschungsgemeinschaft through the grants Ku1315/10-1, TRR60/A2, and SE1885/2-1, the Wilhelm Sander Foundation (2014.136.1), the Deutsche Krebshilfe (70112112), and by the Hariy Cell Leukemia Foundation.

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Authors and Affiliations

  1. Institute of Cell Biology (Cancer Research), Medical Faculty, University of Duisburg-Essen, Virchowstr. 173, 45122, Essen, Germany

    Marc Seifert & Ralf Küppers

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  1. Marc Seifert
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  2. Ralf Küppers
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Correspondence to Ralf Küppers .

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Editors and Affiliations

  1. Immunity and Cancer Laboratory, The Francis Crick Institute, and King’s College London, London, United Kingdom

    Dinis Pedro Calado

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Seifert, M., Küppers, R. (2017). Determining the Origin of Human Germinal Center B Cell-Derived Malignancies. In: Calado, D. (eds) Germinal Centers. Methods in Molecular Biology, vol 1623. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7095-7_20

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  • DOI: https://doi.org/10.1007/978-1-4939-7095-7_20

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  • Publisher Name: Humana Press, New York, NY

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  • Online ISBN: 978-1-4939-7095-7

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