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In vivo imaging of germinal centres reveals a dynamic open structure

Nature volume 446, pages 83–87 (2007)Cite this article

Abstract

Germinal centres are specialized structures wherein B lymphocytes undergo clonal expansion, class switch recombination, antibody gene diversification and affinity maturation. Three to four antigen-specific B cells colonize a follicle to establish a germinal centre and become rapidly dividing germinal-centre centroblasts that give rise to dark zones1,2,3,4. Centroblasts produce non-proliferating centrocytes that are thought to migrate to the light zone of the germinal centre, which is rich in antigen-trapping follicular dendritic cells and CD4+ T cells5,6,7. It has been proposed that centrocytes are selected in the light zone on the basis of their ability to bind cognate antigen5,6,7,8. However, there have been no studies of germinal-centre dynamics or the migratory behaviour of germinal-centre cells in vivo. Here we report the direct visualization of B cells in lymph node germinal centres by two-photon laser-scanning microscopy in mice. Nearly all antigen-specific B cells participating in a germinal-centre reaction were motile and physically restricted to the germinal centre but migrated bi-directionally between dark and light zones. Notably, follicular B cells were frequent visitors to the germinal-centre compartment, suggesting that all B cells scan antigen trapped in germinal centres. Consistent with this observation, we found that high-affinity antigen-specific B cells can be recruited to an ongoing germinal-centre reaction. We conclude that the open structure of germinal centres enhances competition and ensures that rare high-affinity B cells can participate in antibody responses.

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Figure 1: Germinal centre and naive B-cell behaviour.
Figure 2: B-cell behaviour in germinal-centre light and dark zones.
Figure 3: Contacts between B cells and FDC-M1 + cells.
Figure 4: Ongoing seeding of germinal centres.

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Acknowledgements

We thank E. Besmer, M. Zimmer and members of the Nussenzweig laboratory for comments on the manuscript. This work was supported by the Schering Foundation (T.A.S.), a Medical Scientist Training Program grant (R.L.L.), the Rothchild Foundation (G.S.), Fondation de Recherche Medicale (D.S.), and the NIH (M.C.N. and M.L.D.). M.C.N. is an investigator of the Howard Hughes Medical Institute.

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Author notes

  1. Guy Shakhar

    Present address: Present address: Department of Immunology, Weizmann Institute of Science, Rehovot 76100, Israel.,

Authors and Affiliations

  1. Laboratory of Molecular Immunology, and,,

    Tanja A. Schwickert, Randall L. Lindquist, Geulah Livshits, Dimitris Skokos & Michel C. Nussenzweig

  2. Howard Hughes Medical Institute, The Rockefeller University, New York, New York 10021, USA,

    Michel C. Nussenzweig

  3. Program in Molecular Pathogenesis and Department of Pathology, Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, New York 10016, USA,

    Guy Shakhar & Michael L. Dustin

  4. NovImmune SA, 64 avenue de la Roseraie, 1211 Geneva 4, Switzerland,

    Marie H. Kosco-Vilbois

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Correspondence to Michael L. Dustin or Michel C. Nussenzweig.

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Competing interests: Marie Kosco-Vilbois works at Novlmmune, Geneva, as already stated.

Supplementary information

Supplementary Information

This file contains Supplementary Methods, Supplementary Figures S1-S8 and additional references. (PDF 7889 kb)

Supplementary Movie 1

This file contains Supplementary Movie 1 showing a time-lapse movie of a GC without FDC-M1. (MOV 2867 kb)

Supplementary Movie 2

This file contains Supplementary Movie 2 showing a time-lapse movie of GC B cells and Plasmablasts. (MOV 9775 kb)

Supplementary Movie 3

This file contains Supplementary Movie 3 showing a time-lapse movie of a B cell follicle without NP-OVA boost. (MOV 6240 kb)

Supplementary Movie 4

This file contains Supplementary Movie 4 showing a time-lapse movie of a GC. (MOV 6850 kb)

Supplementary Movie 5

This file contains Supplementary Movie 5 showing a two stacked time-lapse movie of a GC. (MOV 10942 kb)

Supplementary Movie 6

This file contains Supplementary Movie 6 showing a time-lapse movie of a WT B cell contacting FDC-M1+ cells. (MOV 3758 kb)

Corrigendum

In vivo imaging of germinal centres reveals a dynamic open structure Tanja A. Schwickert, Randall L. Lindquist, Guy Shakhar, Geulah Livshits, Dimitris Skokos, Marie H. Kosco-Vilbois, Michael L. Dustin and Michel C. Nussenzweig Nature doi:10.1038/nature 05573 (published online 31 January 2007) In Supplementary Movie 4 of this Letter, the LZ and DZ labels were swapped. The corrected version was uploaded on 5 February 2007. (PDF 11 kb)

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Schwickert, T., Lindquist, R., Shakhar, G. et al. In vivo imaging of germinal centres reveals a dynamic open structure. Nature 446, 83–87 (2007). https://doi.org/10.1038/nature05573

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