Direct observation correlates NFκB cRel in B cells with activating and terminating their proliferative program.

Publisher: National Academy of Sciences Country of Publication: United States NLM ID: 7505876 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1091-6490 (Electronic) Linking ISSN: 00278424 NLM ISO Abbreviation: Proc Natl Acad Sci U S A Subsets: MEDLINE

Original Publication: Washington, DC : National Academy of Sciences

B-Lymphocytes*/immunology ; B-Lymphocytes*/metabolism ; Cell Proliferation* ; NF-kappa B*/metabolism; Animals ; Mice ; Mice, Knockout ; Mice, Inbred C57BL ; Proto-Oncogene Proteins c-rel/metabolism ; Proto-Oncogene Proteins c-rel/genetics

Antibody responses require the proliferative expansion of B cells controlled by affinity-dependent signals. Yet, proliferative bursts are heterogeneous, varying between 0 and 8 divisions in response to the same stimulus. NFκB cRel is activated in response to immune stimulation in B cells and is genetically required for proliferation. Here, we asked whether proliferative heterogeneity is controlled by natural variations in cRel abundance. We developed a fluorescent reporter mTFP1-cRel for the direct observation of cRel in live proliferating B cells. We found that cRel is heterogeneously distributed among naïve B cells, which are enriched for high expressors in a heavy-tailed distribution. We found that high cRel expressors show faster activation of the proliferative program, but do not sustain it well, with population expansion decaying earlier. With a mathematical model of the molecular network, we showed that cRel heterogeneity arises from balancing positive feedback by autoregulation and negative feedback by its inhibitor IκBε, confirmed by mouse knockouts. Using live-cell fluorescence microscopy, we showed that increased cRel primes B cells for early proliferation via higher basal expression of the cell cycle driver cMyc. However, peak cMyc induction amplitude is constrained by incoherent feedforward regulation, decoding the fold change of cRel activity to terminate the proliferative burst. This results in a complex nonlinear, nonmonotonic relationship between cRel expression and the extent of proliferation. These findings emphasize the importance of direct observational studies to complement gene knockout results and to learn about quantitative relationships between biological processes and their key regulators in the context of natural variations.
Competing Interests: Competing interests statement:The authors declare no competing interest.

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R01AI132731 HHS | NIH (NIH); N/A The University of Utah, Department of Pathology; N/A James S. McDonnell Foundation (JSMF); R56 AI177789 United States AI NIAID NIH HHS; R01 AI127867 United States AI NIAID NIH HHS; R01AI127867 HHS | NIH (NIH); Scholar Award American Society of Hematology (ASH); DRQ11-21 Damon Runyon Cancer Research Foundation (DRCRF); R01 AI132731 United States AI NIAID NIH HHS

Keywords: B cell proliferation; IκBε negative feedback; cRel fluorescent reporter; cell-to-cell heterogeneity; direct observation

0 (NF-kappa B)
0 (Proto-Oncogene Proteins c-rel)

Date Created: 20240718 Date Completed: 20240718 Latest Revision: 20240816

20240816

PMC11287273

10.1073/pnas.2309686121

39024115