Germinal center reentries of BCL2-overexpressing B cells drive follicular lymphoma progression
Stéphanie Sungalee, … , Bertrand Nadel, Sandrine Roulland
Stéphanie Sungalee, … , Bertrand Nadel, Sandrine Roulland
Published November 10, 2014
Citation Information: J Clin Invest. 2014;124(12):5337-5351. https://doi.org/10.1172/JCI72415.
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Research Article

Germinal center reentries of BCL2-overexpressing B cells drive follicular lymphoma progression

Abstract

It has recently been demonstrated that memory B cells can reenter and reengage germinal center (GC) reactions, opening the possibility that multi-hit lymphomagenesis gradually occurs throughout life during successive immunological challenges. Here, we investigated this scenario in follicular lymphoma (FL), an indolent GC-derived malignancy. We developed a mouse model that recapitulates the FL hallmark t(14;18) translocation, which results in constitutive activation of antiapoptotic protein B cell lymphoma 2 (BCL2) in a subset of B cells, and applied a combination of molecular and immunofluorescence approaches to track normal and t(14;18)+ memory B cells in human and BCL2-overexpressing B cells in murine lymphoid tissues. BCL2-overexpressing B cells required multiple GC transits before acquiring FL-associated developmental arrest and presenting as GC B cells with constitutive activation–induced cytidine deaminase (AID) mutator activity. Moreover, multiple reentries into the GC were necessary for the progression to advanced precursor stages of FL. Together, our results demonstrate that protracted subversion of immune dynamics contributes to early dissemination and progression of t(14;18)+ precursors and shapes the systemic presentation of FL patients.

Authors

Stéphanie Sungalee, Emilie Mamessier, Ester Morgado, Emilie Grégoire, Philip Z. Brohawn, Christopher A. Morehouse, Nathalie Jouve, Céline Monvoisin, Cédric Menard, Guilhaume Debroas, Mustapha Faroudi, Violaine Mechin, Jean-Marc Navarro, Charlotte Drevet, Franziska C. Eberle, Lionel Chasson, Fannie Baudimont, Stéphane J. Mancini, Julie Tellier, Jean-Michel Picquenot, Rachel Kelly, Paolo Vineis, Philippe Ruminy, Bruno Chetaille, Elaine S. Jaffe, Claudine Schiff, Jean Hardwigsen, David A. Tice, Brandon W. Higgs, Karin Tarte, Bertrand Nadel, Sandrine Roulland

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Figure 3

BCL2+ GC and post-GC B cells are able to reenter and reactivate GC reactions during repeated antigenic challenges.

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BCL2+ GC and post-GC B cells are able to reenter and reactivate GC react...
(A) Schematic of the adoptive transfer procedure. (B) Representative F-PCR analysis of BCL2CJ junctions in total spleen and LNs harvested before (left panel) and after adoptive transfer of the indicated EYFP+ cell subpopulations (middle and right panel). BraLN, brachial LN. (C) FACS plots showing the retrieved EYPF+ cells in recipient mouse R1 6 days after adoptive transfer of EYFP+GL7+ cells and F-PCR analysis of BCL2CJ in FACS-sorted EYFP+GL7+ and EYFP+GL7– cells. 650 EYFP+GL7– and 250 EYFP+GL7+ cells were recovered from the EYFP+ cell subpopulation (0.01% of total splenic B cells). (D) Spleen histology from recipient mouse R2 after adoptive transfer of EYFP+GL7–IgM+ cells, stained with IgD (blue), GL7 (white), and huBCL2 (pink) antibodies. huBCL2+ cells in GCs and peri-GCs are shown (arrows). Images are representative of 2 independent experiments with 2 mice per condition. Scale bars: 50 μm (left) and 10 μm (right). (E) Splenic histology from recipient mouse R1 after adoptive transfer of EYFP+GL7+ cells. A single GC invaded with huBCL2+EYFP+ cells was observed among otherwise normal GCs (dashed line outlines), mimicking a human FLIS. Arrows indicate the presence of both GL7+ and GL7– BCL2+EYFP+IgD– cells in the same GC. Images are representative of 3 independent experiments with 2 mice per condition (additional figures in Supplemental Figure 3, A and B). Fo, follicular zone. (F) F-PCR of BCL2CJ in the splenic section containing the FLIS structure that was scraped off the slide. Ten PCR replicates of 100 ng DNA each were performed in parallel, cloned, and sequenced. Scale bars: 200 μm (left) and 30 μm (right).