Germinal center dark and light zone organization is mediated by CXCR4 and CXCR5

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Abstract

Germinal center (GC) dark and light zones segregate cells undergoing somatic hypermutation and antigen-driven selection, respectively, yet the factors guiding this organization are unknown. We report here that GC organization was absent from mice deficient in the chemokine receptor CXCR4. Centroblasts had high expression of CXCR4 and GC B cells migrated toward the CXCR4 ligand SDF-1 (CXCL12), which was more abundant in the dark zone than in the light zone. CXCR4-deficient cells were excluded from the dark zone in the context of a wild-type GC. These findings establish that GC organization depends on sorting of centroblasts by CXCR4 into the dark zone. In contrast, CXCR5 helped direct cells to the light zone and deficiency in CXCL13 was associated with aberrant light zone localization.

Figure 1: CXCR4 deficiency results in GC disorganization.
Figure 2: Treatment with a CXCR4 inhibitor results in GC disorganization.
Figure 3: Deficiency of CXCR4 in B cells disrupts FDC and CXCL13 polarity in the GC.
Figure 4: GC B cells upregulate CXCR4 and show enhanced chemotaxis to SDF-1.
Figure 5: Detection of SDF-1 protein and mRNA in the GC.
Figure 6: CXCR4 is upregulated on centroblasts and is required for dark zone localization.
Figure 7: CXCR5 and CXCL13 function in determining light zone position.

Change history

  • 08 August 2004

    appended erratum PDF to back of AOP PDF; placed footnote in XML at every instance of Fig. 1; corrected online date will appear in issue PDF.

Notes

  1. 1.

    *Note: In the version of this article originally published online, the labels in Figure 1b were positioned incorrectly. This error has been corrected for the HTML and print versions of this article.

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Acknowledgements

We thank D. Hargreaves, Y. Xu and M. Lesneski for technical assistance; C. Miller for training in laser-capture microdissection; J. Dietrich for surgical expertise; S. Jiang for cell sorting; D. Littman for Cxcr4+/− mice; M. Lipp for Cxcr5−/− mice; T. Roach for some Eμ-Bcl2-22 mice; F. Arenzana-Seisdedos for K15C antibody; J. Lin and other members of the Weiss Lab for Jurkat cells; the Werb lab for the use of equipment and supplies; the University of California San Francisco Diabetes Center for use of the ABI Prism 7900HT; S. Luther, T. Okada and G. Cinamon for advice and comments on the manuscript; and M. Matloubian, C. Lo and J. Cholfin for discussions. Work supported by Howard Hughes Medical Institute and grants AI40098 and AI45073 from the National Institutes of Health, and by predoctoral grants from Howard Hughes Medical Institute (C.D.C.A. and K.M.A.).

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Correspondence to Jason G Cyster.

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Supplementary information

Supplementary Fig. 1

GCs in Eμ-bcl-2-22 transgenic mice have normal dark and light zone polarity. (PDF 74 kb)

Supplementary Table 1

Antibodies used in flow cytometry. (PDF 18 kb)

Supplementary Table 2

Primary antibodies used in mouse immunohistochemistry. (PDF 19 kb)

Supplementary Table 3

Secondary antibodies used in mouse immunohistochemistry. (PDF 17 kb)

Supplementary Table 4

Primer and probe sequences for quantitative real-time PCR. (PDF 18 kb)

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Allen, C., Ansel, K., Low, C. et al. Germinal center dark and light zone organization is mediated by CXCR4 and CXCR5. Nat Immunol 5, 943–952 (2004). https://doi.org/10.1038/ni1100

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