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The adaptor protein SLP-65 acts as a tumor suppressor that limits pre-B cell expansion

Nature Immunology volume 4, pages 38–43 (2003)Cite this article

Abstract

Mice deficient in the adaptor protein SLP-65 (also known as BLNK) have reduced numbers of mature B cells, but an increased pre-B cell compartment. We show here that compared to wild-type cells, SLP-65−/− pre-B cells show an enhanced ex vivo proliferative capacity. This proliferation requires interleukin 7 and expression of the pre-B cell receptor (pre-BCR). In addition, SLP-65−/− mice have a high incidence of pre-B cell lymphoma. Reintroduction of SLP-65 into SLP-65−/− pre-B cells led to pre-BCR down-regulation and enhanced differentiation. Our results indicate that SLP-65 regulates a developmental program that promotes differentiation and limits pre-B cell expansion, thereby acting as a tumor suppressor.

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Figure 1: SLP-65−/− BM-derived B cells express large amounts of surface pre-BCR.
Figure 2: The enhanced proliferative capacity of SLP-65−/− BM-derived B cells depends on pre-BCR expression.
Figure 3: The pre-BCR of SLP-65−/− pre-B cell lines is expressed on proliferating cells and is signaling-competent.
Figure 4: Inhibition of Erk activation induces apoptosis in SLP-65−/− pre-B cell lines.
Figure 5: Reconstitution of SLP-65 expression results in down-regulation of surface pre-BCR expression.

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Acknowledgements

We thank P. Nielsen for critically reading the manuscript; A. Wuerch for cell sorting, C. Eschbach and M. Mitterer for technical help; U. Klingmüller for providing retroviral constructs and expert advice; H. Eibel and P. Richter for help in establishing the in vitro culture system. Supported by the Deutsche Forschungs Gemeinschaft through SFB 364 and the Leibniz program.

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  1. Institute for Biology III, Albert-Ludwigs University of Freiburg and Max Planck Institute for Immunobiology, Stuebeweg 51, Freiburg, 79108, Germany

    Alexandra Flemming, Tilman Brummer, Michael Reth & Hassan Jumaa

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Correspondence to Michael Reth or Hassan Jumaa.

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Supplementary information

Web Fig. 1.

The number of κ+ cells increases in SLP-65–/– , but not in WT pre-B cell cultures after IL-7 withdrawal. WT and SLP-65–/– pre-B cells from 1-week cultures were analyzed for κ expression and subsequently plated-out in medium lacking IL-7 for 2 days. The ratio of κ+ cells was determined before and after IL-7 withdrawal. (PDF 22 kb)

Web Fig. 2.

The enhanced proliferation capacity of SLP-65–/– BM-derived B cells depends on pre-BCR expansion. (a) c-Kit versus B220 FACS profiles of lymphocyte-gated cells derived from total BM littermates. (b) Sorted B220+ c-Kit+ cells were cultured in IL-7 (>1 ng/ml). Proliferation was assessed by counting at the indicated time points, values were averaged between duplicates. After 5 days of in vitro culture, the number of SLP-65–/– cells had increased 70-fold, whereas the number of WT cells had only increased 5.5-fold. (PDF 66 kb)

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Flemming, A., Brummer, T., Reth, M. et al. The adaptor protein SLP-65 acts as a tumor suppressor that limits pre-B cell expansion. Nat Immunol 4, 38–43 (2003). https://doi.org/10.1038/ni862

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