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Allelic 'choice' governs somatic hypermutation in vivo at the immunoglobulin κ-chain locus

Nature Immunology volume 8, pages 715–722 (2007)Cite this article

Abstract

Monoallelic demethylation and rearrangement control allelic exclusion of the immunoglobulin κ-chain locus (Igk locus) in B cells. Here, through the introduction of pre-rearranged Igk genes into their physiological position, the critical rearrangement step was bypassed, thereby generating mice producing B cells simultaneously expressing two different immunoglobulin-κ light chains. Such 'double-expressing' B cells still underwent monoallelic demethylation at the Igk locus, and the demethylated allele was the 'preferred' substrate for somatic hypermutation in each cell. However, methylation itself did not directly inhibit the activation-induced cytidine-deaminase reaction in vitro. Thus, it seems that the epigenetic mechanisms that initially bring about monoallelic variable-(diversity)-joining rearrangement continue to be involved in the control of antibody diversity at later stages of B cell development.

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Figure 1: Analysis of splenic lymphocytes.
Figure 2: Methylation analyses of 'double-targeted' mice.
Figure 3: Mutation analysis.
Figure 4: DNA-methylation and expression analyses of Peyer's patch B cells.

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Acknowledgements

We thank I. Keshet for experimental assistance, S. Casola for scientific advice and cell sorting; C. Goettlinger for cell sorting; C. Koenigs, C. Uthoff-Hachenberg and A. Egert for technical help; R. Grützmann (University of Cologne) for anti–mouse κ (R33-18.10) and anti-IgM (R33-24.12). Supported by the National Institutes of Health (H.C., K.R. and Y.B.), the Israel Academy of Sciences (Y.B), the German Israel Foundation (Y.B.), the European Community 5th Framework Quality of Life Program (Y.B.), the Israel Cancer Research Fund (H.C.) and the Volkswagen Foundation (K.R.).

Author information

Author notes

  1. Shira Fraenkel, Raul Mostoslavsky and Tatiana I Novobrantseva: These authors contributed equally to this work.

Authors and Affiliations

  1. The Hebrew University Hadassah Medical School, Jerusalem, 91120, Israel

    Shira Fraenkel, Raul Mostoslavsky, Howard Cedar & Yehudit Bergman

  2. The Center for Blood Research and Department of Genetics, Howard Hughes Medical Institute, The Children's Hospital, Harvard University Medical School, Boston, 02115, Massachusetts, USA

    Raul Mostoslavsky, Jayanta Chaudhuri & Frederick W Alt

  3. Institute for Genetics, University of Cologne, Cologne, D-50931, Germany

    Tatiana I Novobrantseva, Roberta Pelanda & Klaus Rajewsky

  4. Alnylam Pharmaceuticals, Cambridge, 02142, Massachusetts, USA

    Tatiana I Novobrantseva

  5. National Jewish Medical and Research Center, Denver, 80206, Colorado, USA

    Roberta Pelanda

  6. Memorial Sloan Kettering Cancer Center, New York, 10021, New York, USA

    Jayanta Chaudhuri

  7. Artemis Pharmaceuticals, Cologne, 51063, Germany

    Gloria Esposito

  8. Department of Immunology, Weizmann Institute of Science, Rehovot, 76100, Israel

    Steffen Jung

  9. Center for Blood Research, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Klaus Rajewsky

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Correspondence to Yehudit Bergman.

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Competing interests

T.I.N. is an employee of Alnylam Pharmaceuticals and G.E. is an employee of Artemis Pharmaceuticals.

Supplementary information

Supplementary Fig. 1

Generation of the 3-38hCκ and D23mCκ alleles. (PDF 578 kb)

Supplementary Fig. 2

IgM titers in the serum of WT and mutant mice. (PDF 2137 kb)

Supplementary Fig. 3

Flow cytometric analysis of bone marrow lymphocytes. (PDF 1072 kb)

Supplementary Fig. 4

Bisulfite mutation analysis of methylated 3-83 V region molecules. (PDF 838 kb)

Supplementary Fig. 5

CpG methylation does not inhibit AID deamination of flanking Cs. (PDF 2653 kb)

Supplementary Fig. 6

Flow cytometric analyses of splenic lymphocytes from 4-week-old WT:CκT and D23ki:CκT mice. (PDF 509 kb)

Supplementary Table 1

Base exchange pattern of hypermutation. (PDF 10 kb)

Supplementary Table 2

Primer sequences. (PDF 26 kb)

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Fraenkel, S., Mostoslavsky, R., Novobrantseva, T. et al. Allelic 'choice' governs somatic hypermutation in vivo at the immunoglobulin κ-chain locus. Nat Immunol 8, 715–722 (2007). https://doi.org/10.1038/ni1476

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