STAT5 regulates the self-renewal capacity and differentiation of human memory B cells and controls Bcl-6 expression

Abstract

It is unknown how B cells that mature during a germinal center reaction 'decide' between plasma or memory cell fate. Here we describe a previously unknown subpopulation of B cells in the human germinal center that is characterized by tyrosine phosphorylated transcriptional activator STAT5. These cells had an activated centrocyte phenotype and had abundant expression of BCL6 but low expression of PRDM1, both encoding transcriptional repression proteins. Using RNA interference and ectopic expression of constitutively activated forms of STAT5, we demonstrate here a function for STAT5 in the self-renewal of B cells in vitro. STAT5b isoform seemed to directly upregulate Bcl-6, and ectopic expression of Bcl-6 in B cells resulted in self-renewal and inhibition of plasma cell differentiation. These data indicate that activation of STAT5 is involved in regulation of memory B cell differentiation.

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Figure 1: Detection of STAT5 in human B cells in vivo and in vitro.
Figure 2: Quantitative real-time PCR of BCL6 and PRDM1.
Figure 3: Reduction of STAT5 expression limits the proliferative potential of primary human B cells.
Figure 4: Expression of CA-STAT5b leads to survival and expansion of B cell populations, whereas expression of WT-STAT5b results in survival only.
Figure 5: CA-STAT5b-ER localizes to the nucleus and induces B cell population expansion in a 4HT-dependent way.
Figure 6: Characteristics of peripheral blood human B cells transduced with Bcl-6–IRES-GFP and cultured in CD40L, IL-2 and IL-4.
Figure 7: Induction of BCL6 expression by STAT5.

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Acknowledgements

We thank A. Bakker (Netherlands Cancer Institute, Amsterdam, Netherlands) for help in cloning; B. Hooibrink (Academic Medical Center, Amsterdam, Netherlands), A. Pfauth and F. van Diepen (Netherlands Cancer Institute, Amsterdam, Netherlands) for help with cell sorting; T. Dellemijn, L. Oomen and L. Brooks (Netherlands Cancer Institute, Amsterdam, Netherlands) for help with CLSM analyses of the tonsil sections and transduced B cell samples; N. van der Stoep (Leiden University Medical Center, Leiden, Netherlands) for help with the luciferase assays; and the Department of Otolaryngology, Academic Medical Center, Amsterdam, Netherlands (W. Fokkens) for providing tonsil tissue.

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Correspondence to Hergen Spits.

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A patent has been filed (application PCT/5B2002/005753) based on the findings presented here and a company has been founded to commercialize the findings reported here.

Supplementary information

Supplementary Fig. 1

Flow cytometric analysis of a representative culture of CA-STAT5b-ER-IRES-ΔNGFR transduced B cells cultured in CD40L, IL-2 and IL-4 in the presence of 4HT. (PDF 219 kb)

Supplementary Fig. 2

Expression of WT-STAT5b-ER in IgG+ cells results in a 4HT dependent expansion in the presence of CD40L, IL-2 and IL-4. (PDF 83 kb)

Supplementary Fig. 3

Effect of BCL6 knock down on the growth of CA-STAT5b transduced B cells. (PDF 88 kb)

Supplementary Fig. 4

Activation of CA-STAT5-ER by 4HT upregulates MYC in transduced primary CD19+ cells. (PDF 106 kb)

Supplementary Fig. 5

Activation of CA-STAT5-ER by 4HT upregulates PAX5 in transduced primary CD19+ cells. (PDF 137 kb)

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Scheeren, F., Naspetti, M., Diehl, S. et al. STAT5 regulates the self-renewal capacity and differentiation of human memory B cells and controls Bcl-6 expression. Nat Immunol 6, 303–313 (2005). https://doi.org/10.1038/ni1172

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