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Cannabinoid receptor 2 mediates the retention of immature B cells in bone marrow sinusoids

Nature Immunology volume 10, pages 403–411 (2009)Cite this article

Abstract

Immature B cells developing in the bone marrow are found in the parenchyma and sinusoids. The mechanisms that control the positioning of B cells in the sinusoids are not understood. Here we show that the integrin α4β1 (VLA-4) and its ligand VCAM-1 were required, whereas the chemokine receptor CXCR4 was dispensable, for sinusoidal retention of B cells. Instead, cannabinoid receptor 2 (CB2), a Gαi protein–coupled receptor upregulated in immature B cells, was required for sinusoidal retention. Using two-photon microscopy, we found immature B cells entering and crawling in sinusoids; these immature B cells were displaced by CB2 antagonism. Moreover, CB2-deficient mice had a lower frequency of immunoglobulin λ-chain–positive B cells in the peripheral blood and spleen. Our findings identify unique requirements for the retention of B cells in the bone marrow sinusoidal niche and suggest involvement of CB2 in the generation of the B cell repertoire.

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Figure 1: In vivo labeling of B lymphocytes in bone marrow sinusoids.
Figure 2: Retention of B cells in bone marrow sinusoids is promoted by α4 and VCAM-1.
Figure 3: Retention of B lymphocytes in bone marrow sinusoids by β1 integrins.
Figure 4: CXCR4 contributes to the retention of B lineage cells in bone marrow parenchyma but does not account for sinusoidal retention.
Figure 5: CB2 is required for the lodgment of immature B cells in bone marrow sinusoids.
Figure 6: Migration dynamics of immature B cells in bone marrow sinusoids.
Figure 7: Constitutive requirement for CB2 in the retention of B cells in bone marrow sinusoids.
Figure 8: Lower frequency of λ+ B cells in CB2-deficient mice.

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Acknowledgements

We thank N. Sakaguchi (Kumamoto University) for Rag1GFP/GFP mice; T. Gerdes and M. Wabl (University of California San Francisco) for Rag1GFP/+ mice; H. Tamamura and N. Fujii (Kyoto University) for the CXCR4 antagonist; Y.-R. Zhou (Columbia University) and D. Littman (New York University) for CXCR4-deficient mice; K. Rajewsky (CBR Institute, Harvard Medical School) for Cd19Cre/+ mice; C. Allen for technical help and discussions; L. Shiow for discussions; I. Grigorova for technical help and discussions; and S. Vilarinho, T. Arnon, E. Gray and S.R. Schwab for comments on the manuscript. Supported by the Howard Hughes Medical Institute (J.P.P. and J.G.C.) and the National Institutes of Health (AI40098).

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Authors and Affiliations

  1. Howard Hughes Medical Institute and Department of Microbiology and Immunology, University of California San Francisco, San Francisco, California, USA

    João P Pereira, Jinping An, Ying Xu & Jason G Cyster

  2. Department of Biopharmaceutical Sciences, Drug Studies Unit, University of California San Francisco, South San Francisco, California, USA

    Yong Huang

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Contributions

J.P.P. did the experiments; J.A. maintained the mouse colonies; Y.X. did the quantitative PCR analysis; Y.H. did the mass spectrometry analysis; and J.P.P. and J.G.C. designed and conceptualized the research, analyzed the data and prepared the manuscript.

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Correspondence to Jason G Cyster.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4 and Supplementary Methods (PDF 685 kb)

Supplementary Video 1

Immature B cells enter and crawl inside BM sinusoids. Timelapse sequence of 51μm z-projection images of the BM calvarium of an anesthetized adult Rag1GFP/+ mouse. A cell in parenchyma and a cell inside a sinusoid and their migratory paths were highlighted with a flashing white arrowhead, and with white dots, respectively. The flashing yellow arrowhead and yellow dots draw attention to a cell in the parenchyma entering and crawling inside a sinusoid. Elapsed time is shown in mm:ss. (MOV 6084 kb)

Supplementary Video 2

Immature B cell arrest and crawling inside BM sinusoids. Timelapse sequence of 51μm z-projection images of the BM calvarium of an anesthetized adult Rag1GFP/+ mouse. Two cells inside sinusoids are highlighted with flashing yellow arrowheads moments before being released and disappearing in flow. The white arrowhead indicates the region where a cell arrests and begins crawling inside the sinsuoid. The white dots depict the migratory path of the cell. Elapsed time is shown in mm:ss. (MOV 8807 kb)

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Pereira, J., An, J., Xu, Y. et al. Cannabinoid receptor 2 mediates the retention of immature B cells in bone marrow sinusoids. Nat Immunol 10, 403–411 (2009). https://doi.org/10.1038/ni.1710

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