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Nonequivalent nuclear location of immunoglobulin alleles in B lymphocytes

Nature Immunology volume 2, pages 848–854 (2001)Cite this article

Abstract

Individual B lymphocytes normally express immunoglobulin (Ig) proteins derived from single Ig heavy chain (H) and light chain (L) alleles. Allelic exclusion ensures monoallelic expression of Ig genes by each B cell to maintain single receptor specificity. Here we provide evidence that at later stages of B cell development, additional mechanisms may contribute to prioritizing expression of single IgH and IgL alleles. Fluorescent in situ hybridization analysis of primary splenic B cells isolated from normal and genetically manipulated mice showed that endogenous IgH, κ and λ alleles localized to different subnuclear environments after activation and had differential expression patterns. However, this differential recruitment and expression of Ig alleles was not typically seen among transformed B cell lines. These data raise the possibility that epigenetic factors help maintain the monoallelic expression of Ig.

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Figure 1: Organization of the IgH and IgL loci.
Figure 2: Nuclear position of IgH, κ and λ genes relative to Ikaros protein and centromeric DNA clusters in activated splenic B lymphocytes.
Figure 3: RNA-FISH analysis of nuclear IgH-, Igκ- and CD45-derived transcripts generated after the activation of splenic B cells.
Figure 4: Rearrangement and RNA expression of IgH alleles in pre- and pro-B cell clones.
Figure 5: Nuclear location of productive and nonproductively rearranged IgH alleles in transformed 70Z/3 and 38C-13 cell lines.

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Acknowledgements

We thank our colleagues for supplying mouse genomic probes; S. Smale for anti-Ikaros sera; G. Klaus for anti-IgM; J. Roes for Cμ-Cδ probes; P. Brodeur for VHJ558 and VHS107 probes; E. Severinson, L. Ström, V. Buckle and N. Brockdorff for helpful discussions; and G. Reed, R. Newton and I. Devonish for photographic and secretarial assistance. Supported by the Medical Research Council, the Wellcome Trust (J. S. and D. G.) and the Royal Society (K. E. B.).

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Authors and Affiliations

  1. Lymphocyte Development Group, MRC Clinical Sciences Centre, Imperial College School of Medicine, Hammersmith Hospital, Du Cane Road, London, W12 ONN, UK

    Jane A. Skok, Karen E. Brown, Veronique Azuara, Marie-Laure Caparros, Jonathan Baxter, Katalin Takacs, Matthias Merkenschlager & Amanda G. Fisher

  2. Gene Regulation and Chromatin Group, MRC Clinical Sciences Centre, Imperial College School of Medicine, Hammersmith Hospital, Du Cane Road, London, W12 ONN, UK

    Niall Dillon

  3. Institute of Cell, Animal and Population Biology, University of Edinburgh, Ashworth Laboratories, Kings Buildings, West Mains Road, Edinburgh, EH9 3JT, UK

    David Gray

  4. Institute for Cancer Research, Fox Chase Cancer Center, 7701 Burholme Avenue, Philadelphia, 19111, PA, USA

    Robert P. Perry

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Correspondence to Amanda G. Fisher.

Supplementary information

Web Figure 1.

Measurement of allelic distances between IgH loci and γ-satellite DNA. (a) Sequential images (ordered from the top left to right, middle left to right, lower left to right) of optical sections through the nucleus of a 4-day-activated mouse B cell nucleus (Z series, 0.2 µm separation) in which the location of IgH alleles (green) and γ-satellite DNA (red) is shown. Data from 54 individual cells was subjected to 3D reconstruction (using Metamorph Imaging series 4.5, Universal Imaging Corporation) to allow 3-D distances between pixels in X, Y and Z planes to be estimated. (b,c) Intranuclear distances (in mm) between the centroid (signal intensity weighted center) of the nearest γ-satellite signal and the center of each IgH allele (maximum intensity). (b) Distribution analysis in which the distance of the nearest (allele 1) and furthest (allele 2) IgH allele from centromeric DNA, is given. (c) Distances were plotted for each individual cell. (GIF 55 kb)

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Skok, J., Brown, K., Azuara, V. et al. Nonequivalent nuclear location of immunoglobulin alleles in B lymphocytes. Nat Immunol 2, 848–854 (2001). https://doi.org/10.1038/ni0901-848

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