title: Visualization of RelB expression and activation at the single-cell level during dendritic cell maturation in Relb-Venus knock-in mice.
abstract: RelB is activated by the non-canonical NF-κB pathway, which is crucial for immunity by establishing lymphoid organogenesis and B-cell and dendritic cell (DC) maturation. To elucidate the mechanism of the RelB-mediated immune cell maturation, a precise understanding of the relationship between cell maturation and RelB expression and activation at the single-cell level is required. Therefore, we generated knock-in mice expressing a fusion protein between RelB and fluorescent protein (RelB-Venus) from the Relb locus. The Relb(Venus/Venus) mice developed without any abnormalities observed in the Relb(-/-) mice, allowing us to monitor RelB-Venus expression and nuclear localization as RelB expression and activation. Relb(Venus/Venus) DC analyses revealed that DCs consist of RelB(-), RelB(low) and RelB(high) populations. The RelB(high) population, which included mature DCs with projections, displayed RelB nuclear localization, whereas RelB in the RelB(low) population was in the cytoplasm. Although both the RelB(low) and RelB(-) populations barely showed projections, MHC II and co-stimulatory molecule expression were higher in the RelB(low) than in the RelB(-) splenic conventional DCs. Taken together, our results identify the RelB(low) population as a possible novel intermediate maturation stage of cDCs and the Relb(Venus/Venus) mice as a useful tool to analyse the dynamic regulation of the non-canonical NF-κB pathway.
authors.affiliation: Division of Cellular and Molecular Biology, Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan;
authors.forename: Takao
authors.initials: T
authors.lastname: Seki
authors.affiliation: Division of Cellular and Molecular Biology, Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan;
authors.forename: Mami
authors.initials: M
authors.lastname: Yamamoto
authors.affiliation: Division of Cellular and Molecular Biology, Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan;
authors.forename: Yuu
authors.initials: Y
authors.lastname: Taguchi
authors.affiliation: Division of Cellular and Molecular Biology, Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan;
authors.forename: Maki
authors.initials: M
authors.lastname: Miyauchi
authors.affiliation: Division of Cellular and Molecular Biology, Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan;
authors.forename: Nobuko
authors.initials: N
authors.lastname: Akiyama
authors.affiliation: Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675, Japan; and.
authors.forename: Noritaka
authors.initials: N
authors.lastname: Yamaguchi
authors.affiliation: Research Center for Asian Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan.
authors.forename: Jin
authors.initials: J
authors.lastname: Gohda
authors.affiliation: Division of Cellular and Molecular Biology, Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan;
authors.forename: Taishin
authors.initials: T
authors.lastname: Akiyama
authors.affiliation: Division of Cellular and Molecular Biology, Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan; jun-i@ims.u-tokyo.ac.jp.
authors.forename: Jun-ichiro
authors.initials: J
authors.lastname: Inoue
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism; D051940:B7-2 Antigen / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology*; D005260:Female; D005786:Gene Expression Regulation; D055879:Gene Knock-In Techniques; D005802:Genes, MHC Class II; D017930:Genes, Reporter; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D015398:Signal Transduction; D059010:Single-Cell Analysis; D013154:Spleen / Q000166:cytology; D013950:Thymus Gland / Q000166:cytology; D051841:Transcription Factor RelB / Q000235:genetics / Q000378:metabolism*
chemical_list: D051940:B7-2 Antigen; D001426:Bacterial Proteins; D008164:Luminescent Proteins; C497594:Relb protein, mouse; C054562:yellow fluorescent protein, Bacteria; D051841:Transcription Factor RelB
keywords: NF-κB; RelB; dendritic cell; single cell; venus