In-vitro derived germinal centre B cells differentially generate memory B or plasma cells in vivo

Nojima, Toshio; Haniuda, Kei; Moutai, Tatsuya; Matsudaira, Moeko; Mizokawa, Sho; Shiratori, Ikuo; Azuma, Takachika; Kitamura, Daisuke

doi:10.1038/ncomms1475

Cited by 264 publications

(379 citation statements)

References 59 publications

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“…5). B1a-derived iGB cells showed localization similar to that of B2-derived iGB cells as reported previously (45). Because iGB-cultured cells are unselected for Ag specificity, they presumably migrate into the extrafollicular region.…”

Section: Discussionsupporting

confidence: 52%

“…Feeder cells expressing CD40L and BAFF (40LB) were prepared as described previously (45). After 4 d of culture, the cells were transferred onto new feeder cells with 10 ng/ml IL-21 (PeproTech) and cultured for 3 d in the presence or absence of 10 mg/ml JNK inhibitor SP600125 (Wako, Osaka, Japan).…”

Section: In Vitro Gc B Cell Culturementioning

confidence: 99%

“…To this end, we used an in vitro culture system that supports long-term growth of GC B cells (iGB) on fibroblasts expressing CD40L and BAFF (45), and observed the cells in the GC area after adoptive transfer in vivo. As a result, the B1a cells that were highly enriched from the peritoneal cavity of BWF1 and G5PR Tg mice differentiated into GC-like B cells.…”

mentioning

confidence: 99%

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JNK Regulatory Molecule G5PR Induces IgG Autoantibody–Producing Plasmablasts from Peritoneal B1a Cells

Kitabatake

Soma

Zhang

et al. 2015

The Journal of Immunology

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Peritoneal B1a cells expressing CD5 and CD11b generate autoantibody-producing precursors in autoimmune-prone mice. Previous studies show reduced JNK signaling in peritoneal B1a cells of female New Zealand Black mice and an abnormal increase of protein phosphatase 2A subunit G5PR that regulates BCR-mediated JNK signaling as a cause of autoimmunity. To investigate the mechanism regulating B1a differentiation into autoantibody-secreting plasmablasts (PBs), we applied an in vitro culture system that supports long-term growth of germinal center (GC) B cells (iGB) with IL-4, CD40L, and BAFF. Compared with spleen B2 cells, B1a cells differentiated into GC-like B cells, but more markedly into PBs, and underwent class switching toward IgG1. During iGB culture, B1a cells expressed GC-associated aicda, g5pr, and bcl6, and markedly PB-associated prdm1, irf4, and xbp1. B1a-derived iGB cells from New Zealand Black × New Zealand White F1 mice highly differentiated into autoantibody-secreting PBs in vitro and localized to the GC area in vivo. In iGB culture, JNK inhibitor SP600125 augmented the differentiation of C57BL/6 B1a cells into PBs. Furthermore, B1a cells from G5PR transgenic mice markedly differentiated into IgM and IgG autoantibody–secreting PBs. In conclusion, JNK regulation is critical to suppress autoantibody-secreting PBs from peritoneal B1a cells.

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Section: Discussionsupporting

confidence: 52%

Section: In Vitro Gc B Cell Culturementioning

confidence: 99%

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JNK Regulatory Molecule G5PR Induces IgG Autoantibody–Producing Plasmablasts from Peritoneal B1a Cells

Kitabatake

Soma

Zhang

et al. 2015

The Journal of Immunology

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“…Retroviral Plat-E packaging cells were maintained in DMEM (Gibco) supplied with 10% (vol/vol) FCS (Gibco), 2 mM L-glutamine (Gibco), and 2 mM sodium pyruvate (Gibco). 40LB feeder cells, producing BAFF and CD40L, were previously generated by Nojima et al (17) and maintained in completed DMEM. To prepare the feeder layer, 40LB feeder cells were irradiated with 12 Gy and plated at 5 × 10 4 cells per centimeter.…”

Section: Methodsmentioning

confidence: 99%

Efficient CRISPR-mediated mutagenesis in primary immune cells using CrispRGold and a C57BL/6 Cas9 transgenic mouse line

Chu

Graf

Wirtz

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

119

136

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International audienceApplying clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9)-mediated mutagenesis to primary mouse immune cells, we used high-fidelity single guide RNAs (sgRNAs) designed with an sgRNA design tool (CrispRGold) to target genes in primary B cells, T cells, and macrophages isolated from a Cas9 transgenic mouse line. Using this system, we achieved an average knockout efficiency of 80% in B cells. On this basis, we established a robust small-scale CRISPR-mediated screen in these cells and identified genes essential for B-cell activation and plasma cell differentiation. This screening system does not require deep sequencing and may serve as a precedent for the application of CRISPR/Cas9 to primary mouse cells

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“…iGB cell culture and iMB cell generation iGB cell culture and iMB cell formation were performed as previously described (42). In brief, purified B cells from CD45.1 + mice were cultured for 4 d on irradiated 40LB cell feeder layer with rIL-4 (1 ng/ml; Peprotech).…”

Section: Preparation Of 3t3 Cells Expressing Integrin a V Bmentioning

confidence: 99%

gp49B-Mediated Negative Regulation of Antibody Production by Memory and Marginal Zone B Cells

Fukao

Haniuda

Nojima

et al. 2014

The Journal of Immunology

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The rapid Ab responses observed after primary and secondary immunizations are mainly derived from marginal zone (MZ) and memory B cells, respectively, but it is largely unknown how these responses are negatively regulated. Several inhibitory receptors have been identified and their roles have been studied, but mainly on follicular B cells and much less so on MZ B, and never on memory B cells. gp49B is an Ig superfamily member that contains two ITIMs in its cytoplasmic tail, and it has been shown to negatively regulate mast cell, macrophage, and NK cell responses. In this study, we demonstrate that gp49B is preferentially expressed on memory and MZ B cells. We show that gp49B−/− mice produce more IgM after a primary immunization and more IgM and IgG1 after a secondary immunization than gp49B+/+ mice in T cell–dependent immune responses. Memory and MZ B cells from gp49B−/− mice also produce more Abs upon in vitro stimulation with CD40 than those from gp49B+/+ mice. The in vitro IgM production by MZ B cells from gp49B+/+, but not gp49B−/−, mice is suppressed by interaction with a putative gp49B ligand, the integrin αvβ3 heterodimer. In addition, gp49B−/− mice exhibited exaggerated IgE production in the memory recall response. These results suggest that plasma cell development from memory and MZ B cells, as well as subsequent Ab production, are suppressed via gp49B. In memory B cells, this suppression also prevents excessive IgE production, thus curtailing allergic diseases.

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In-vitro derived germinal centre B cells differentially generate memory B or plasma cells in vivo

Cited by 264 publications

References 59 publications

JNK Regulatory Molecule G5PR Induces IgG Autoantibody–Producing Plasmablasts from Peritoneal B1a Cells

JNK Regulatory Molecule G5PR Induces IgG Autoantibody–Producing Plasmablasts from Peritoneal B1a Cells

Efficient CRISPR-mediated mutagenesis in primary immune cells using CrispRGold and a C57BL/6 Cas9 transgenic mouse line

gp49B-Mediated Negative Regulation of Antibody Production by Memory and Marginal Zone B Cells

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