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The proto-oncogene MYC is required for selection in the germinal center and cyclic reentry

Nature Immunology volume 13, pages 1083–1091 (2012)Cite this article

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Abstract

After antigenic challenge, B cells enter the dark zone (DZ) of germinal centers (GCs) to proliferate and hypermutate their immunoglobulin genes. Mutants with greater affinity for the antigen are positively selected in the light zone (LZ) to either differentiate into plasma and memory cells or reenter the DZ. The molecular circuits that govern positive selection in the GC are not known. We show here that the GC reaction required biphasic regulation of expression of the cell-cycle regulator c-Myc that involved its transient induction during early GC commitment, its repression by Bcl-6 in DZ B cells and its reinduction in B cells selected for reentry into the DZ. Inhibition of c-Myc in vivo led to GC collapse, which indicated an essential role for c-Myc in GCs. Our results have implications for the mechanism of GC selection and the role of c-Myc in lymphomagenesis.

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Figure 1: A subset of LZ GC B cells express c-Myc under physiological conditions.
Figure 2: Alternating peaks of c-Myc expression and Bcl-6 expression during T cell–dependent antigen responses and GC formation.
Figure 3: Bcl-6 represses c-Myc protein expression in DZ GC B cells.
Figure 4: Coordinated upregulation of genes of the 'immune activation' signature and those encoding molecules involved in entry into the cell cycle in GFP–c-Myc+ GC B cells.
Figure 5: The BCR repertoire of GFP–c-Myc+ GC B cells shows enrichment for high-affinity variants.
Figure 6: Access to T cell help triggers c-Myc expression before reentry into the DZ.
Figure 7: The biological activity of c-Myc is required for normal GC maintenance.

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Acknowledgements

We thank B. Sleckman (Washington University) for GFP–c-Myc mice; S. Artandi (Stanford University) for mice with transgenic expression of rtTA-actin; G. Evan (University of Cambridge, UK) and L. Soucek (Vall d'Hebron Institute of Oncology) for TRE-Omomyc rtTA-Actin mice; L. Reavie, I. Aifantis (New York University), M. Chesi and L. Bergsagel (Mayo Clinic) for access to these mice; F.W. Alt for advice on immunization protocols; R. Bosch for experimental help; K. Basso, A.B. Holmes, L. Pasqualucci, M. Bansal, C. Lefebvre, P. Sumazin and A. Califano for advice on data analysis; T. Mo for mouse husbandry; H. Tang, Q. Shen, V. Miljkovic, K. Gordon, C. Liu, and S. Tetteh for technical assistance; S. Serrano (Parc de Salut Mar, Barcelona) for human tissue samples; M. Jara-Acevedo for technical advice; and U. Klein for insights and for critical reading of this manuscript. Supported by the US National Institutes of Health (P01-CA092625 and R01-CA37295 to R.D.-F., and R01-AI037526 to M.C.N.), the Leukemia and Lymphoma Society (R.D.-F.), the National Cancer Institute (K99/R00-CA151827 to D.D.-S.), the Whitehead Institute for Biomedical Research (G.D.V.) and the Howard Hughes Medical Institute (M.C.N.).

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Author notes

  1. Gabriel D Victora, Ryan T Phan & Masumichi Saito

    Present address: Present addresses: Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA (G.D.V.), Department of Medicine, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, USA (R.T.P.), and Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan (M.S.).,

  2. David Dominguez-Sola and Gabriel D Victora: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute for Cancer Genetics, Columbia University, New York, New York, USA

    David Dominguez-Sola, Carol Y Ying, Ryan T Phan, Masumichi Saito & Riccardo Dalla-Favera

  2. Laboratory of Molecular Immunology, The Rockefeller University, New York, New York, USA

    Gabriel D Victora & Michel C Nussenzweig

  3. Howard Hughes Medical Institute, New York, New York, USA

    Michel C Nussenzweig

  4. Herbert Irving Comprehensive Cancer Center, Columbia University, New York, New York, USA

    Riccardo Dalla-Favera

  5. Department of Pathology and Cell Biology, Columbia University, New York, New York, USA

    Riccardo Dalla-Favera

  6. Department of Genetics and Development, Columbia University, New York, New York, USA

    Riccardo Dalla-Favera

  7. Department of Microbiology and Immunology, Columbia University, New York, New York, USA

    Riccardo Dalla-Favera

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Contributions

D.D.-S. and G.D.V designed the study, did the experiments, analyzed the data and wrote the manuscript; D.D.-S. did the computational analyses; C.Y.Y. did experiments; C.Y.Y., R.T.P. and M.S. identified and characterized the Bcl-6–c-Myc transcriptional interaction; R.D.-F. and M.C.N. supervised the project, provided direction in the study design and wrote the manuscript; and all authors approved the final manuscript.

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Correspondence to Michel C Nussenzweig or Riccardo Dalla-Favera.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–7 and Tables 4–6 (PDF 4310 kb)

Supplementary Table 1

BCL6 ChIP-on-chip binding profile across the proximal region of the MYC locus. (XLSX 50 kb)

Supplementary Table 2

GFPMYC signature genes. (XLSX 31 kb)

Supplementary Table 3

Gene Set Enrichment Analysis of GFPMYC signatures. (XLSX 22 kb)

Supplementary Table 7

List of primers used in real-time quantitative RT-PCR (qRT-PCR), quantitative chromatin immunoprecipitation (qChIP) and IgH-V gene sequence analysis. (XLS 51 kb)

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Dominguez-Sola, D., Victora, G., Ying, C. et al. The proto-oncogene MYC is required for selection in the germinal center and cyclic reentry. Nat Immunol 13, 1083–1091 (2012). https://doi.org/10.1038/ni.2428

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