The sphingosine 1-phosphate receptor S1P2 maintains the homeostasis of germinal center B cells and promotes niche confinement

Abstract

Mice deficient in sphingosine 1-phosphate receptor type 2 (S1P2) develop diffuse large B cell lymphoma. However, the role of S1P2 in normal germinal center (GC) physiology is unknown. Here we show that S1P2-deficient GC B cells outgrew their wild-type counterparts in chronically established GCs. We found that antagonism of the kinase Akt mediated by S1P2 and its downstream mediators Gα12, Gα13 and p115RhoGEF regulated cell viability and was required for growth control in chronically proliferating GCs. Moreover, S1P2 inhibited GC B cell responses to follicular chemoattractants and helped confine cells to the GC. In addition, S1P2 overexpression promoted the centering of activated B cells in the follicle. We suggest that by inhibiting Akt activation and migration, S1P2 helps restrict GC B cell survival and localization to an S1P-low niche at the follicle center.

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Figure 1: Growth advantage of GC B cells deficient in S1P2, Gα12-Gα13 or p115RhoGEF in chronic GCs.
Figure 2: Resistance to apoptosis and greater Akt activation in GC B cells deficient in S1P2, Gα12-Gα13 or p115RhoGEF.
Figure 3: Akt activation confers an advantage to mucosal GCs, and S1P2 regulates translation in GC cells.
Figure 4: Regulation of the migration and positioning of GC B cells by S1P2.
Figure 5: S1P2 acts together with CXCR5 and FDCs to promote GC B cell clustering.
Figure 6: Degradation of S1P by B cells.
Figure 7: S1P2 directs activated B cells to the GC and the center of the follicle.

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Acknowledgements

We thank N. Danial (Dana-Farber) for Bad-deficient bone marrow; K. Rajewsky (Children's Hospital and Immune Disease Institute, Harvard) for mice expressing the Cr2-Cre transgene; J. Browning (Biogen Idec) for lymphotoxin-β receptor–Fc fusion protein; J. An for help with the mouse colony; and S. Schwab for critical reading of the manuscript. Supported by the National Science Foundation (J.A.G.), Howard Hughes Medical Institute (J.G.C.) and the US National Institutes of Health (Intramural Research Program; National Institute of Diabetes and Digestive and Kidney Diseases (R.L.P.); HL65590 and HL44907 to S.R.C.; and AI45073 to J.G.C.).

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Authors

Contributions

J.A.G., B.C. and J.G.C. designed the experiments; J.A.G. did most of the experiments; B.C. did some of the early experiments; K.S. and J.A.G. did the imaging experiments; C.D.C.A. and Y.X. did the initial gene-expression analysis; T.H.S. and J.A.G. did the experiments with PTEN inhibitor; L.D.W. and S.R.C. generated Gα12-deficient mice; D.P. and S.R.C. generated Gna12−/−Gna13f/fMx1-Cre bone marrow; R.L.P. generated S1P2-deficient mice; J.A.G., K.S., B.C. and J.G.C. analyzed the data; and J.A.G. and J.G.C. wrote the paper.

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Correspondence to Jason G Cyster.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–8 (PDF 1607 kb)

41590_2011_BFni2047_MOESM12_ESM.mpg

Movement of S1pr2+/+ Hy10 GC B cells within FDC network. (MPG 5540 kb)

Supplementary Video 1

Movement of S1pr2+/+ Hy10 GC B cells within FDC network. (MPG 5540 kb)

41590_2011_BFni2047_MOESM13_ESM.mpg

Movement of S1pr2−/− Hy10 GC B cells around the perimeter of the FDC network. (MPG 5490 kb)

Supplementary Video 2

Movement of S1pr2−/− Hy10 GC B cells around the perimeter of the FDC network. (MPG 5490 kb)

41590_2011_BFni2047_MOESM14_ESM.mpg

Tracks of wild-type and S1pr2−/− Hy10 GC B cells in and around PE-IC-labeled FDC network. (MPG 20721 kb)

Supplementary Video 3

Tracks of wild-type and S1pr2−/− Hy10 GC B cells in and around PE-IC-labeled FDC network. (MPG 20721 kb)

41590_2011_BFni2047_MOESM15_ESM.mpg

Movement of S1pr2+/+ Hy10 GC B cells within a GC surrounded by an excess of labeled follicular B cells. (MPG 9826 kb)

Supplementary Video 4

Movement of S1pr2+/+ Hy10 GC B cells within a GC surrounded by an excess of labeled follicular B cells. (MPG 9826 kb)

41590_2011_BFni2047_MOESM16_ESM.mpg

Movement of S1pr2−/− Hy10 GC B cells around the perimeter of a GC surrounded by an excess of labeled follicular B cells. (MPG 8424 kb)

Supplementary Video 5

Movement of S1pr2−/− Hy10 GC B cells around the perimeter of a GC surrounded by an excess of labeled follicular B cells. (MPG 8424 kb)

41590_2011_BFni2047_MOESM17_ESM.mpg

Tracks of wild-type and S1pr2−/− Hy10 GC B cells with respect to the surface of the GC, defined by the distribution of wild-type follicular and GC B cells. (MPG 25770 kb)

Supplementary Video 6

Tracks of wild-type and S1pr2−/− Hy10 GC B cells with respect to the surface of the GC, defined by the distribution of wild-type follicular and GC B cells. (MPG 25770 kb)

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Green, J., Suzuki, K., Cho, B. et al. The sphingosine 1-phosphate receptor S1P2 maintains the homeostasis of germinal center B cells and promotes niche confinement. Nat Immunol 12, 672–680 (2011). https://doi.org/10.1038/ni.2047

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