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Transcription factor IRF4 controls plasma cell differentiation and class-switch recombination

Nature Immunology volume 7, pages 773–782 (2006)Cite this article

Abstract

B cells producing high-affinity antibodies are destined to differentiate into memory B cells and plasma cells, but the mechanisms leading to those differentiation pathways are mostly unknown. Here we report that the transcription factor IRF4 is required for the generation of plasma cells. Transgenic mice with conditional deletion of Irf4 in germinal center B cells lacked post–germinal center plasma cells and were unable to differentiate memory B cells into plasma cells. Plasma cell differentiation required IRF4 as well as the transcriptional repressor Blimp-1, which both acted 'upstream' of the transcription factor XBP-1. In addition, IRF4-deficient B cells had impaired expression of activation-induced deaminase and lacked class-switch recombination, suggesting an independent function for IRF4 in this process. These results identify IRF4 as a crucial transcriptional 'switch' in the generation of functionally competent plasma cells.

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Figure 1: Functional analysis of the conditionally deleted Irf4 allele.
Figure 2: Development of GC B cells in Irf4fl/−Cγ1-Cre mice.
Figure 3: Irf4fl/−Cγ1-Cre mice lack plasma cells.
Figure 4: Response of Irf4−/− B cells to LPS stimulation.
Figure 5: Response of Irf4−/− B cells to stimulation with CD40 plus IL-4.
Figure 6: Generation of antigen-specific memory B cells by Irf4fl/−Cγ1-Cre mice.
Figure 7: Lack of memory–to–plasma cell differentiation in Irf4fl/−Cγ1-Cre mice.

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Acknowledgements

We thank P. Smith and M. Benito for help with the immunohistochemistry; M. Deren for genotyping; J. Gao for help with ELISA; K. Gordon and S. Stefanova for cell sorting; V. Miljkovic for sequencing; M. Alizhamov for pEasyFlox; D. Tarlinton (Walter Eliza Hall Institute, Parkville, Australia) for advice on the detection of NP-binding B cells; and L. Pasqualucci, M. Saito and A. Pernis for discussions. Supported by the National Institutes of Health (CA92625 and CA098285 to K.R. and CA92625 and CA37295 to R.D.-F.) and the Human Frontiers Science Program (U.K.).

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Authors and Affiliations

  1. Department of Pathology and Herbert Irving Comprehensive Cancer Center, Institute for Cancer Genetics, Columbia University, New York, 10032, New York, USA

    Ulf Klein, Giorgio Cattoretti, Qiong Shen, Marie Lia, Tongwei Mo, Thomas Ludwig & Riccardo Dalla-Favera

  2. CBR Institute for Biomedical Research, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Stefano Casola & Klaus Rajewsky

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Contributions

U.K. designed research, did experiments, analyzed data and wrote the manuscript; R.D.-F. designed research and wrote the manuscript; S.C. and K.R. contributed the Cγ1-Cre mouse, which was critical for this analysis, and designed research; G.C. did the immunostaining experiments and analyzed data; Q.S. did the embryonic stem cell injection and ELISA; M.L. did the VH sequencing analysis; T.M. maintained and genotyped the mouse cohort; and T.L. was involved in construction of the transgenic Irf4 mouse and provided materials.

Note: Supplementary information is available on the Nature Immunology website.

Corresponding author

Correspondence to Riccardo Dalla-Favera.

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Supplementary information

Supplementary Fig. 1

Generation and functional analysis of the conditional Irf4 allele (PDF 109 kb)

Supplementary Fig. 2

Functionality of the conditional Irf4 allele in vivo. (PDF 280 kb)

Supplementary Fig. 3

Absence of plasma cells in Irf4f1/−Cγ1-Cre mice. (PDF 51 kb)

Supplementary Fig. 4

Production of IgG1+ cells in B cell cultures from Irf4fl/−Cγ1-Cre mice stimulated with CD40 plus IL-4 and CSFE analysis of CD40 plus IL-4-stimulated Irf4−/− cells. (PDF 159 kb)

Supplementary Fig. 5

IgG1+eGFP+ B cells in Irf4fl/−Cγ1-Cre mice display a phenotype typical of memory B cells. (PDF 72 kb)

Supplementary Fig. 6

Lack of memory to plasma cell differentiation in Irf4fl/−Cγ1-Cre mice. (PDF 99 kb)

Supplementary Table 1

Sequence analysis of γ1 (membrane-form) transcripts amplified splenic and PB B cells of Irf4fl/−, Irf4fl/+ and Irf4+/− mice at day 14 after NP-KLH immunization. (PDF 23 kb)

Supplementary Table 2

Sequence analysis of γ1 transcripts amplified from B cell purified from Irf4fl/−Cγ1-Cre and Irf4fl/+Cγ1-Cre mice at day 14 after NP-KLH immunization. (PDF 25 kb)

Supplementary Table 3

Sequence analysis of γ1 transcripts amplified from purified PB B cells of Irf4fl/−Cγ1-Cre and Irf4fl/+Cγ1-Cre mice at day 42 after NP-KLH immunization. (PDF 23 kb)

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Klein, U., Casola, S., Cattoretti, G. et al. Transcription factor IRF4 controls plasma cell differentiation and class-switch recombination. Nat Immunol 7, 773–782 (2006). https://doi.org/10.1038/ni1357

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