Article | Published:

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

Nature Immunology volume 8, pages 715722 (2007) | Download Citation

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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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

    • Shira Fraenkel
    • , Raul Mostoslavsky
    •  & Tatiana I Novobrantseva

    These authors contributed equally to this work.

Affiliations

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

    • Shira Fraenkel
    • , Raul Mostoslavsky
    • , Howard Cedar
    •  & Yehudit Bergman
  2. Howard Hughes Medical Institute, The Children's Hospital, The Center for Blood Research and Department of Genetics, Harvard University Medical School, Boston, Massachusetts 02115, 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, Massachusetts 02142, USA.

    • Tatiana I Novobrantseva
  5. National Jewish Medical and Research Center, Denver, Colorado 80206, USA.

    • Roberta Pelanda
  6. Memorial Sloan Kettering Cancer Center, New York, New York 10021, 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, Massachusetts 02115, USA.

    • Klaus Rajewsky

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Competing interests

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

Corresponding author

Correspondence to Yehudit Bergman.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Generation of the 3-38hCκ and D23mCκ alleles.

  2. 2.

    Supplementary Fig. 2

    IgM titers in the serum of WT and mutant mice.

  3. 3.

    Supplementary Fig. 3

    Flow cytometric analysis of bone marrow lymphocytes.

  4. 4.

    Supplementary Fig. 4

    Bisulfite mutation analysis of methylated 3-83 V region molecules.

  5. 5.

    Supplementary Fig. 5

    CpG methylation does not inhibit AID deamination of flanking Cs.

  6. 6.

    Supplementary Fig. 6

    Flow cytometric analyses of splenic lymphocytes from 4-week-old WT:CκT and D23ki:CκT mice.

  7. 7.

    Supplementary Table 1

    Base exchange pattern of hypermutation.

  8. 8.

    Supplementary Table 2

    Primer sequences.

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DOI

https://doi.org/10.1038/ni1476

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