Thiranut Jaroonwitchawan, Hideki Arimochi, Yuki Sasaki, Chieko Ishifune, Hiroyuki Kondo, Kunihiro Otsuka, Shin-ichi Tsukumo and Koji Yasutomo : Stimulation of the farnesoid X receptor promotes M2 macrophage polarization., Frontiers in Immunology, Vol.14, 2023.
(Summary)
This skewed polarization towards M2 macrophages by an FXR agonist was accompanied by increased expression of signaling molecules related to the retinoic acid receptor. Inhibition of the retinoic acid receptor suppressed FXR agonist-mediated M2 macrophage polarization, indicating that this polarization was, at least, partly dependent on the retinoic acid receptor pathway. These data demonstrate that FXR has a role in polarization toward M2 macrophages and suggest a possible therapeutic potential of FXR agonists in M2 macrophage-related conditions.
Shin-ichi Tsukumo, Ganesh Poorani Subramani, Noé Seija, Mizuho Tabata, Yoichi Maekawa, Yuya Mori, Chieko Ishifune, Yasushi Itoh, Mineto Ota, Keishi Fujio, M Noia Javier Di and Koji Yasutomo : AFF3, a susceptibility factor for autoimmune diseases, is a molecular facilitator of immunoglobulin class switch recombination., Science Advances, Vol.8, No.34, 2022.
(Summary)
in human B cells. These findings demonstrate that AFF3 directly regulates CSR by facilitating the recruitment of AID to the switch regions.
Chieko Ishifune, Shin-ichi Tsukumo, Yoichi Maekawa, Katsuto Hozumi, Hyun Doo Chung, Chihiro Motozono, Sho Yamasaki, Hiroyasu Nakano and Koji Yasutomo : Regulation of membrane phospholipid asymmetry by Notch-mediated flippase expression controls the number of intraepithelial TCRαβ+CD8αα+ T cells., PLoS Biology, Vol.17, No.5, 2019.
(Summary)
Intestinal intraepithelial lymphocytes (IELs) expressing CD8αα on αβ T cells (TCRαβ+CD8αα+ IELs) have suppressive capabilities in enterocolitis, but the mechanism that maintains homeostasis and cell number is not fully understood. Here, we demonstrated that the number of TCRαβ+CD8αα+ IELs was severely reduced in mice lacking recombination signal binding protein for immunoglobulin kappa J region (Rbpj) or Notch1 and Notch2 in T cells. Rbpj-deficient TCRαβ+CD8αα+ IELs expressed low levels of Atp8a2, which encodes a protein with flippase activity that regulates phospholipid asymmetry of plasma membrane such as flipping phosphatidylserine in the inner leaflet of plasma membrane. Rbpj-deficient TCRαβ+CD8αα+ IELs cannot maintain phosphatidylserine in the inner leaflet of the plasma membrane. Furthermore, depletion of intestinal macrophages restored TCRαβ+CD8αα+ IELs in Rbpj-deficient mice, suggesting that exposure of phosphatidylserine on the plasma membrane in Rbpj-deficient TCRαβ+CD8αα+ IELs acts as an "eat-me" signal. Together, these results revealed that Notch-Atp8a2 is a fundamental regulator for IELs and highlighted that membrane phospholipid asymmetry controlled by Notch-mediated flippase expression is a critical determinant in setting or balancing the number of TCRαβ+CD8αα+ IELs.
MicroRNAs have broad roles in tumorigenesis and cell differentiation through regulation of target genes. Notch signaling also controls cell differentiation and tumorigenesis. However, the mechanisms through which Notch mediates microRNA expression are still unclear. In this study, we aimed to identify microRNAs regulated by Notch signaling. Our analysis found that microRNA-449a (miR-449a) was indirectly regulated by Notch signaling. Although miR-449a-deficient mice did not show any Notch-dependent defects in immune cell development, treatment of miR-449a-deficient mice with azoxymethane (AOM) or dextran sodium sulfate (DSS) increased the numbers and sizes of colon tumors. These effects were associated with an increase in intestinal epithelial cell proliferation following AOM/DSS treatment. In patients with colon cancer, miR-449a expression was inversely correlated with disease-free survival and histological scores and was positively correlated with the expression of MLH1 for which loss-of function mutations have been shown to be involved in colon cancer. Colon tissues of miR-449a-deficient mice showed reduced Mlh1 expression compared with those of wild-type mice. Thus, these data suggested that miR-449a acted as a key regulator of colon tumorigenesis by controlling the proliferation of intestinal epithelial cells. Additionally, activation of miR-449a may represent an effective therapeutic strategy and prognostic marker in colon cancer.
Sultana Taskia Zaman, Hideki Arimochi, Satoshi Maruyama, Chieko Ishifune, Shin-ichi Tsukumo, Akiko Kitamura and Koji Yasutomo : Notch Balances Th17 and Induced Regulatory T Cell Functions in Dendritic Cells by Regulating Aldh1a2 Expression., The Journal of Immunology, Vol.199, No.6, 1989-1997, 2017.
(Summary)
Dendritic cells (DCs) are important for adaptive immune responses through the activation of T cells. The molecular interplay between DCs and T cells determines the magnitude of T cell responses or outcomes of functional differentiation of T cells. In this study, we demonstrated that DCs in mice that are Rbpj deficient in CD11c(+) cells (Rbpj(-/-) mice) promoted the differentiation of IL-17A-producing Th17 cells. Rbpj-deficient DCs expressed little Aldh1a2 protein that is required for generating retinoic acid. Those DCs exhibited a reduced ability for differentiating regulatory T cells induced by TGF-. Rbpj protein directly regulated Aldh1a2 transcription by binding to its promoter region. The overexpression of Aldh1a2 in Rbpj-deficient DCs negated their Th17-promoting ability. Transfer of naive CD4(+) T cells into Rag1-deficient Rbpj(-/-) mice enhanced colitis with increased Th17 and reduced induced regulatory T cells (iTreg) compared with control Rag1-deficient mice. The cotransfer of iTreg and naive CD4(+) T cells into Rag1-deficient Rbpj(-/-) mice improved colitis compared with transfer of naive CD4(+) T cell alone. Furthermore, cotransfer of DCs from Rbpj(-/-) mice that overexpressed Aldh1a2 or Notch-stimulated DCs together with naive CD4(+) T cells into Rbpj(-/-)Rag1-deficient mice led to reduced colitis with increased iTreg numbers. Therefore, our studies identify Notch signaling in DCs as a crucial balancer of Th17/iTreg, which depends on the direct regulation of Aldh1a2 transcription in DCs.
Takahiro Furukawa, Chieko Ishifune, Shin-ichi Tsukumo, Katsuto Hozumi, Yoichi Maekawa, Naoko Matsui, Ryuji Kaji and Koji Yasutomo : Transmission of survival signals through Delta-like 1 on activated CD4(+) T cells., Scientific Reports, Vol.6, 33692, 2016.
(Summary)
Notch expressed on CD4(+) T cells transduces signals that mediate their effector functions and survival. Although Notch signaling is known to be cis-inhibited by Notch ligands expressed on the same cells, the role of Notch ligands on T cells remains unclear. In this report we demonstrate that the CD4(+) T cell Notch ligand Dll1 transduces signals required for their survival. Co-transfer of CD4(+) T cells from Dll1(-/-) and control mice into recipient mice followed by immunization revealed a rapid decline of CD4(+) T cells from Dll1(-/-) mice compared with control cells. Dll1(-/-) mice exhibited lower clinical scores of experimental autoimmune encephalitis than control mice. The expression of Notch target genes in CD4(+) T cells from Dll1(-/-) mice was not affected, suggesting that Dll1 deficiency in T cells does not affect cis Notch signaling. Overexpression of the intracellular domain of Dll1 in Dll1-deficient CD4(+) T cells partially rescued impaired survival. Our data demonstrate that Dll1 is an independent regulator of Notch-signaling important for the survival of activated CD4(+) T cells, and provide new insight into the physiological roles of Notch ligands as well as a regulatory mechanism important for maintaining adaptive immune responses.
Jaba Gamrekelashvili, Roberto Giagnorio, Jasmin Jussofie, Oliver Soehnlein, Johan Duchene, G Carlos Briseño, K Saravana Ramasamy, Kashyap Krishnasamy, Anne Limbourg, Tamar Kapanadze, Chieko Ishifune, Rabea Hinkel, Freddy Radtke, J Lothar Strobl, Ursula Zimber-Strobl, Christian L Napp, Johann Bauersachs, Hermann Haller, Koji Yasutomo, Christian Kupatt, M Kenneth Murphy, H Ralf Adams, Christian Weber and P Florian Limbourg : Regulation of monocyte cell fate by blood vessels mediated by Notch signalling., Nature Communications, Vol.7, 12597, 2016.
(Summary)
A population of monocytes, known as Ly6C(lo) monocytes, patrol blood vessels by crawling along the vascular endothelium. Here we show that endothelial cells control their origin through Notch signalling. Using combinations of conditional genetic deletion strategies and cell-fate tracking experiments we show that Notch2 regulates conversion of Ly6C(hi) monocytes into Ly6C(lo) monocytes in vivo and in vitro, thereby regulating monocyte cell fate under steady-state conditions. This process is controlled by Notch ligand delta-like 1 (Dll1) expressed by a population of endothelial cells that constitute distinct vascular niches in the bone marrow and spleen in vivo, while culture on recombinant DLL1 induces monocyte conversion in vitro. Thus, blood vessels regulate monocyte conversion, a form of committed myeloid cell fate regulation.
SJ Oh, S Ahn, YH Jin, Chieko Ishifune, JH Kim, Koji Yasutomo and DH Chung : Notch 1 and Notch 2 synergistically regulate the differentiation and function of invariant NKT cells., Journal of Leukocyte Biology, Vol.98, No.5, 781-789, 2015.
(Summary)
Invariant natural killer T cells are a distinct subset of T cells that exert Janus-like functions. Moreover, Notch signaling is known to have critical roles in the development and functions of T cells. However, it is not known whether Notch signaling contributes to the development or functions of invariant natural killer T cells. Here, we found that CD4-specific gene ablation of Notch 1 and Notch 2 (N1N2(-/-)) increased the number of invariant natural killer T cells in the thymus but decreased them in the liver. N1N2(-/-) mice showed impaired thymic maturation of invariant natural killer T cells from the NK1.1(-)CD44(+) to the NK1.1(+)CD44(+) stage, resulting in accumulation of NK1.1(-)CD44(+) invariant natural killer T cells in the thymus. Upon activation, hepatic invariant natural killer T cells from N1N2(-/-) mice produced lower cytokine levels and increased apoptosis versus wild-type invariant natural killer T cells. Furthermore, Notch 1/Notch 2-deficient, but not wild type, invariant natural killer T cells failed to promote antibody-induced arthritis in CD1d(-/-) mice. Unlike N1N2(-/-) mice, RBP-j(lox) (/) (lox) CD4-Cre mice showed similar percentages and numbers of thymic invariant natural killer T cells to wild-type mice but had defects in their homeostasis, maturation, and cytokine production in the liver. Taken together, our data indicate distinct effects of Notch signaling on invariant natural killer T cells in the thymus and liver, which are at least partly independent of RBP-j in the thymus.
Kurihara Takeshi, Hideki Arimochi, Bhuyan Ahmed Zaied, Chieko Ishifune, Tsumura Hideki, Ito Morihiro, Ito Yasuhiko, Akiko Kitamura, Yoichi Maekawa and Koji Yasutomo : CD98 Heavy Chain Is a Potent Positive Regulator of CD4+ T Cell Proliferation and Interferon- Production In Vivo, PLoS ONE, Vol.10, No.10, e0139692, 2015.
(Summary)
Upon their recognition of antigens presented by the MHC, T cell proliferation is vital for clonal expansion and the acquisition of effector functions, which are essential for mounting adaptive immune responses. The CD98 heavy chain (CD98hc, Slc3a2) plays a crucial role in the proliferation of both CD4+ and CD8+ T cells, although it is unclear if CD98hc directly regulates the T cell effector functions that are not linked with T cell proliferation in vivo. Here, we demonstrate that CD98hc is required for both CD4+ T cell proliferation and Th1 functional differentiation. T cell-specific deletion of CD98hc did not affect T cell development in the thymus. CD98hc-deficient CD4+ T cells proliferated in vivo more slowly as compared with control T cells. C57BL/6 mice lacking CD98hc in their CD4+ T cells could not control Leishmania major infections due to lowered IFN- production, even with massive CD4+ T cell proliferation. CD98hc-deficient CD4+ T cells exhibited lower IFN- production compared with wild-type T cells, even when comparing IFN- expression in cells that underwent the same number of cell divisions. Therefore, these data indicate that CD98hc is required for CD4+ T cell expansion and functional Th1 differentiation in vivo, and suggest that CD98hc might be a good target for treating Th1-mediated immune disorders.
Yoichi Maekawa, Chieko Ishifune, Shin-ichi Tsukumo, Katsuto Hozumi, Hideo Yagita and Koji Yasutomo : Notch controls the survival of memory CD4+ T cells by regulating glucose uptake., Nature Medicine, Vol.21, No.1, 55-61, 2015.
(Summary)
CD4+ T cells differentiate into memory T cells that protect the host from subsequent infection. In contrast, autoreactive memory CD4+ T cells harm the body by persisting in the tissues. The underlying pathways controlling the maintenance of memory CD4+ T cells remain undefined. We show here that memory CD4+ T cell survival is impaired in the absence of the Notch signaling protein known as recombination signal binding protein for immunoglobulin J region (Rbpj). Treatment of mice with a Notch inhibitor reduced memory CD4+ T cell numbers and prevented the recurrent induction of experimental autoimmune encephalomyelitis. Rbpj-deficient CD4+ memory T cells exhibit reduced glucose uptake due to impaired AKT phosphorylation, resulting in low Glut1 expression. Treating mice with pyruvic acid, which bypasses glucose uptake and supplies the metabolite downstream of glucose uptake, inhibited the decrease of autoimmune memory CD4+ T cells in the absence of Notch signaling, suggesting memory CD4+ T cell survival relies on glucose metabolism. Together, these data define a central role for Notch signaling in maintaining memory CD4+ T cells through the regulation of glucose uptake.
(Keyword)
Animals / CD4-Positive T-Lymphocytes / Glucose / Glucose Transporter Type 1 / Humans / Immunoglobulin J Recombination Signal Sequence-Binding Protein / Mice / Oncogene Protein v-akt / Receptors, Antigen, T-Cell / Receptors, Notch / Signal Transduction
Chieko Ishifune, Satoshi Maruyama, Yuki Sasaki, Hideo Yagita, Katsuto Hozumi, Taisuke Tomita, Kenji Kishihara and Koji Yasutomo : Differentiation of CD11c+ CX3CR1+ cells in the small intestine requires Notch signaling., Proceedings of the National Academy of Sciences of the United States of America, Vol.111, No.16, 5986-5991, 2014.
(Summary)
The gastrointestinal tract comes into direct contact with environmental agents, including bacteria, viruses, and foods. Intestine-specific subsets of immune cells maintain gut homeostasis by continuously sampling luminal antigens and maintaining immune tolerance. CD11c(+)CX3CR1(+) cells sample luminal antigens in the small intestine and contribute to the trafficking of bacteria to lymph nodes under dysbiotic conditions. The molecular mechanisms crucial for the differentiation of CD11c(+)CX3CR1(+) cells remain unclear. Here we demonstrate that the Notch1- or Notch2-Rbpj axis is essential for the development of CD11c(+)CX3CR1(+) cells. In mice in which Rbpj or Notch1 and Notch2 were deleted from CD11c(+) cells, there was a deficit of CD11c(+)CX3CR1(+) cells and an accumulation of CD11c(low)CX3CR1(+) cells. The CD11c(low)CX3CR1(+) cells could not differentiate to CD11c(+)CX3CR1(+) cells, suggesting that CD11c(low)CX3CR1(+) cells represent a lineage distinct from CD11c(+)CX3CR1(+) cells. These data indicate that Notch signaling is essential for lineage fixation of intestinal CD11c(+)CX3CR1(+) cells.
(Keyword)
Animals / Antigens, CD11c / Cell Count / Cell Differentiation / Immunoglobulin J Recombination Signal Sequence-Binding Protein / Intestine, Small / Mice / Mice, Inbred C57BL / Receptors, Chemokine / Receptors, Notch / Signal Transduction
Ahmed Zaied Bhuyan, Michihito Asanoma, Akiko Iwata, Chieko Ishifune, Yoichi Maekawa, Mitsuo Shimada and Koji Yasutomo : Abrogation of Rbpj attenuates experimental autoimmune uveoretinitis by inhibiting IL-22-producing CD4+ T cells., PLoS ONE, Vol.9, No.2, e89266, 2014.
(Summary)
Experimental autoimmune uveoretinitis (EAU) is an organ-specific T cell-mediated disease induced by immunizing mice with interphotoreceptor retinoid binding protein (IRBP). Autoaggressive CD4(+) T cells are the major pathogenic population for EAU. We investigated the contribution of Notch signaling in T cells to EAU pathogenesis because Notch signaling regulates various aspects of CD4(+) T cell functions. Rbpj is required for Notch signaling, and Rbpj deficiency in T cells inhibited EAU disease severity. The amelioration of EAU in T cell-specific Rbpj-deficient mice correlated with low levels of IL-22 production from CD4(+) T cells, although IRBP-specific CD4(+) T cell proliferation and Th17 differentiation were unaffected. Administration of recombinant IL-22 during the late phase, but not the early phase, of EAU increased EAU clinical scores in T cell-specific Rbpj-deficient mice. Notch inhibition in mice immunized with IRBP with a -secretase inhibitor (GSI) suppressed EAU progression, even when GSI was administered as late as 13 days after IRBP immunization. Our data demonstrate that Rbpj/Notch-mediated IL-22 production in T cells has a key pathological role in the late phase of EAU, and suggest that Notch blockade might be a useful therapeutic approach for treating EAU.
Ahmed Zaied Bhuyan, Hideki Arimochi, Jun Nishida, Keiko Kataoka, Takeshi Kurihara, Chieko Ishifune, Hideki Tsumura, Morihiro Ito, Yasuhiko Ito, Akiko Kitamura and Koji Yasutomo : CD98hc regulates the development of experimental colitis by controlling effector and regulatory CD4(+) T cells., Biochemical and Biophysical Research Communications, Vol.444, No.4, 628-633, 2014.
(Summary)
CD4(+) T cell activation is controlled by signaling through the T cell receptor in addition to various co-receptors, and is also affected by their interactions with effector and regulatory T cells in the microenvironment. Inflammatory bowel diseases (IBD) are caused by the persistent activation and expansion of auto-aggressive CD4(+) T cells that attack intestinal epithelial cells. However, the molecular basis for the persistent activation of CD4(+) T cells in IBD remains unclear. In this study, we investigated how the CD98 heavy chain (CD98hc, Slc3a2) affected the development of colitis in an experimental animal model. Transferring CD98hc-deficient CD4(+)CD25(-) T cells into Rag2(-/-) mice did not cause colitis accompanied by increasing Foxp3(+) inducible regulatory T cells. By comparison, CD98hc-deficient naturally occurring regulatory T cells (nTregs) had a decreased capability to suppress colitis induced by CD4(+)CD25(-) T cells, although CD98hc-deficient mice did not have a defect in the development of nTregs. Blocking CD98hc with an anti-CD98 blocking antibody prevented the development of colitis. Our results indicate that CD98hc regulates the expansion of autoimmune CD4(+) T cells in addition to controlling nTregs functions, which suggests the CD98hc as an important target molecule for establishing strategies for treating colitis.
Kohhei Nakajima, Yoichi Maekawa, Keiko Kataoka, Chieko Ishifune, Jun Nishida, Hideki Arimochi, Akiko Kitamura, Takayuki Yoshimoto, Shuhei Tomita, Shinji Nagahiro and Koji Yasutomo : The ARNT-STAT3 axis regulates the differentiation of intestinal intraepithelial TCRαβ+CD8αα+cells., Nature Communications, Vol.4, 2112, 2013.
(Summary)
Intestinal intraepithelial T cells contribute to the regulation of inflammatory responses in the intestine; however, the molecular basis for their development and maintenance is unknown. The aryl hydrocarbon receptor complexes with the aryl hydrocarbon receptor nuclear translocator (ARNT) and senses environmental factors, including gut microbiota. Here, we identify ARNT as a critical regulator of the differentiation of TCRαβ(+)CD8αα(+) intestinal intraepithelial T cells. Mice deficient in either ARNT or aryl hydrocarbon receptor show a greater than- eight-fold reduction in the number of TCRαβ(+)CD8αα(+) intestinal intraepithelial T cells. The number of TCRαβ(+)CD8αα(+) intestinal intraepithelial T cells is increased by treatment with an aryl hydrocarbon receptor agonist in germ-free mice and is decreased by antibiotic treatment. The Arnt-deficient precursors of TCRαβ(+)CD8αα(+) intestinal intraepithelial T cells express low amounts of STAT3 and fail to differentiate towards the TCRαβ(+)CD8αα(+) cell fate after IL-15 stimulation, a deficiency that is overcome by overexpression of Stat3. These data demonstrate that the ARNT-STAT3 axis is a critical regulator of TCRαβ(+)CD8αα(+) intestinal intraepithelial T-cell development and differentiation.
Shuichi Iwahashi, Yoichi Maekawa, Jun Nishida, Chieko Ishifune, Akiko Kitamura, Hideki Arimochi, Keiko Kataoka, Shigeru Chiba, Mitsuo Shimada and Koji Yasutomo : Notch2 regulates the development of marginal zone B cells through Fos., Biochemical and Biophysical Research Communications, Vol.418, No.4, 701-707, 2012.
(Summary)
B cells are classified into several subsets depending on their functions, marker expression pattern and localization. Marginal zone B (MZB) cells are a distinct lineage from follicular B cells, and regulate host defenses against blood-borne pathogens. Notch2/RBP-J signaling regulates the development of MZB cells by interacting with delta-like 1 ligand, although the target genes for Notch2 signaling remain unclear. We identified Fos as an upregulated gene in LPS-stimulated B cells that received Notch2 signaling. Fos is expressed in CD21(high)CD23(low) MZB cells at a higher level compared to CD21(Int)CD23(high) follicular B cells. Deleting the Notch2 gene in CD19(+) B cells decreased Fos expression in B cells. Overexpression of Fos in Notch2-deficient B cells or bone marrow cells partially restored MZB development. Fos promoter activity was upregulated by Notch2 signaling, indicating that Notch2 directly controls Fos transcription associated with MZB development. These data identify Fos as one of the target genes for Notch2 signaling that is crucial for MZB development.
Tomomi Kuwahara, Yositoshi Ogura, Kenshiro Oshima, Ken Kurokawa, Tadasuke Ooka, Hideki Hirakawa, Takehiko Itoh, Haruyuki Nakayama-Imaohji, Minoru Ichimura, Kikuji Itoh, Chieko Ishifune, Yoichi Maekawa, Koji Yasutomo, Masahira Hattori and Tetsuya Hayashi : The lifestyle of the segmented filamentous bacterium: a non-culturable gut-associated immunostimulating microbe inferred by whole-genome sequencing., DNA Research, Vol.18, No.4, 291-303, 2011.
(Summary)
Numerous microbes inhabit the mammalian intestinal track and strongly impact host physiology; however, our understanding of this ecosystem remains limited owing to the high complexity of the microbial community and the presence of numerous non-culturable microbes. Segmented filamentous bacteria (SFBs), which are clostridia-related Gram-positive bacteria, are among such non-culturable populations and are well known for their unique morphology and tight attachment to intestinal epithelial cells. Recent studies have revealed that SFBs play crucial roles in the post-natal maturation of gut immune function, especially the induction of Th17 lymphocytes. Here, we report the complete genome sequence of mouse SFBs. The genome, which comprises a single circular chromosome of 1 620 005 bp, lacks genes for the biosynthesis of almost all amino acids, vitamins/cofactors and nucleotides, but contains a full set of genes for sporulation/germination and, unexpectedly, for chemotaxis/flagella-based motility. These findings suggest a triphasic lifestyle of the SFB, which comprises two types of vegetative (swimming and epicellular parasitic) phases and a dormant (spore) phase. Furthermore, SFBs encode four types of flagellin, three of which are recognized by Toll-like receptor 5 and could elicit the innate immune response. Our results reveal the non-culturability, lifestyle and immunostimulation mechanisms of SFBs and provide a genetic basis for the future development of the SFB cultivation and gene-manipulation techniques.
Chieko Ishifune, Yoichi Maekawa, Jun Nishida, Akiko Kitamura, Tanigaki Kenji, Yagita Hideo and Koji Yasutomo : Notch signaling regulates the development of a novel type of Thy1-expressing dendritic cell in the thymus., European Journal of Immunology, Vol.41, No.5, 1309-1320, 2011.
(Summary)
Dendritic cells (DCs) are specialized antigen-presenting cells (APCs) required for T-cell activation and are classified into several subtypes by phenotypic and functional characteristics. However, it remains unclear if distinct transcription factors control the development of each DC subpopulation. In this report, we demonstrate that Notch signaling controls the development of a novel DC subtype that expresses Thy1 (Thy1(+) DCs). Overstimulation of bone marrow cells with the Notch ligand Delta-like 1 promoted the development of Thy1(+) DCs. Thy1(+) DCs are characterized as CD11c(+) MHC class II(+) NK1.1(-) B220(-) CD8(+) , and are present in the thymus but not in the spleen and lymph nodes. Thymic Thy1(+) DCs are able to capture exogenous proteins and delete CD4(+) CD8(+) T cells. Transplantation experiments demonstrated that CD44(+) CD25(-) and CD44(+) CD25(+) thymocytes can differentiate into Thy1(+) DCs. Recombination signal binding protein for immunoglobulin kappa J region (RBP-J) deficiency in lineage-negative bone marrow cells, but not CD11c(+) cells, disrupted Thy1(+) DC development in the thymus. Our data indicate that Notch signaling controls the development of a novel type of Thy1-expressing DC in the thymus that possibly controls negative selection, and indicates that there may be highly regulated, differential transcriptional control of DC development. Furthermore, our findings suggest that Notch signaling regulates T-cell development not only by intrinsically inducing T-cell lineage-specific gene programs, but also by regulating negative selection through Thy1(+) DCs.
Yoichi Maekawa, Yoshiaki Minato, Chieko Ishifune, Takeshi Kurihara, Akiko Kitamura, Hidefumi Kojima, Hideo Yagita, Mamiko Sakata-Yanagimoto, Toshiki Saito, Ichiro Taniuchi, Shigeru Chiba, Saburo Sone and Koji Yasutomo : Notch2 integrates signaling by the transcription factors RBP-J and CREB1 to promote T cell cytotoxicity., Nature Immunology, Vol.9, No.10, 1140-1147, 2008.
(Summary)
The acquisition of cytotoxic effector function by CD8(+) T cells is crucial for the control of intracellular infection and tumor invasion. However, it remains unclear which signaling pathways are required for the differentiation of CD8(+) cytotoxic T lymphocytes. We show here that Notch2-deficient T cells had impaired differentiation into cytotoxic T lymphocytes. In addition, dendritic cells with lower expression of the Notch ligand Delta-like 1 induced the differentiation of cytotoxic T lymphocytes less efficiently. We found that the intracellular domain of Notch2 interacted with a phosphorylated form of the transcription factor CREB1, and together these proteins bound the transcriptional coactivator p300 to form a complex on the promoter of the gene encoding granzyme B. Our results suggest that the highly regulated, dynamic control of T cell cytotoxicity depends on the integration of Notch2 and CREB1 signals.
Mika Kijima, Takeshi Yamaguchi, Chieko Ishifune, Yoichi Maekawa, Akemi Koyanagi, HIdeo Yagita, Shigeru Chiba, Kenji Kishihara, Mitsuo Shimada and Koji Yasutomo : Dendritic cell-mediated NK cell activation is controlled by Jagged2-Notch interaction., Proceedings of the National Academy of Sciences of the United States of America, Vol.105, No.19, 7010-7015, 2008.
(Summary)
Natural killer (NK) cells regulate various immune responses by exerting cytotoxic activity or secreting cytokines. The interaction of NK cells with dendritic cells (DC) contributes to NK cell-mediated antitumor or antimicrobial responses. However, the cellular and molecular mechanisms for controlling this interaction are largely unknown. Here, we show an involvement of Jagged2-Notch interaction in augmenting NK cell cytotoxicity mediated by DC. Enforced expression of Jagged2 on A20 cells (Jag2-A20 cells) suppressed their growth in vivo, which was abrogated by depleting NK cells. Moreover, Jag2-A20 cells exerted a suppression on the growth of nonmanipulated A20 cells in SCID mice in an NK-dependent manner. Consistently, coinoculation of A20 cells with DC overexpressing Jagged2 (Jag2-DC) suppressed the growth of A20 cells in mice. Stimulation of NK cells with Jagged2 directly enhanced their cytotoxicity, IFN-gamma production, and proliferation. Ligation of Notch2 on NK cells enhanced their cytotoxic activity, and Jag2-DC or CpG-treated DC-mediated NK cell cytotoxicity was suppressed by a gamma-secretase inhibitor. These results indicate that the Jagged2-Notch axis plays a crucial role in DC-mediated NK cell cytotoxicity. Furthermore, manipulation of this interaction may provide an approach to induce potent tumor immunity or to inhibit certain autoimmune diseases caused by NK cell activation.
Chieko Ishifune and Koji Yasutomo : Ahrによる腸管の恒常性制御 (レドックスUPDATE ストレス制御の臨床医学・健康科学に関する別冊号), Journal of Clinical and Experimental Medicine, 90-95, Jul. 2015.
2.
Chieko Ishifune and Koji Yasutomo : The differentiation of intestinal dendtitic cells and the Notch signaling, Clinical Immunology & Allergology, Vol.58, No.4, 382-386, Oct. 2012.
3.
Chieko Ishifune and 安友 康二 : IL-22 and the New IL-10 Family (IL-19, 20, 22, 24, 26), 細胞工学, Vol.31, No.7, 751-757, Jun. 2012.
Chieko Ishifune and Koji Yasutomo : NotchによるPARP1活性化はB細胞特異的に腫瘍化を抑制する, Hematology, Vol.63, No.6, 734-739, Dec. 2011.
5.
Chieko Ishifune and Koji Yasutomo : Regulation of T cell homeostasis by Notch signaling, Clinical Immunology & Allergology, Vol.55, No.5, 490-495, May 2011.
(Keyword)
Notch / T cell / homeostasis / proliferation / differentiation
Chieko Ishifune and Koji Yasutomo : 【樹状細胞による免疫制御と臨床応用 T細胞制御機構の理解から，樹状細胞療法の開発，自己免疫疾患・感染症の病態解明とその治療まで】 樹状細胞の生物学と生理学(分化・サブセット活性化・動態) Notchシグナルと樹状細胞分化, Experimental Medicine, Vol.26, No.20, 3169-3174, Dec. 2008.
7.
Chieko Ishifune and Koji Yasutomo : 【制御性T細胞の多様性と臨床応用】 微生物感染と制御性T細胞, Inflammation & Immunology, Vol.15, No.4, 437-442, Jul. 2007.
Proceeding of International Conference:
1.
Chieko Ishifune and Koji Yasutomo : Notch signal controls TCR+CD8+ intestinal intraepithelial lymphocyte, 7th Joint Meeting of Tokushima University, Gifu University, KAIST, and Seoul National University, Seoul, Nov. 2018.
2.
Chieko Ishifune, Shin-ichi Tsukumo and Koji Yasutomo : Notch signal controls the number of TCR+CD8+ intraepithelial lymphocytes via phospholipid asymmetry by maintaining flippase ATP8a2, Australia-Japan Meeting on Cell Death, Tokyo, May 2018.
3.
Chieko Ishifune, Yoichi Maekawa, Katsuto Hozumi and Koji Yasutomo : Notch-STAT5b is central for the development of TCR alpha beta+ CD8 alpha alpha+ small intestinal intraepithelial lymphocytes, 16th International Congress of Immunology (ICI), Melbourne, Australia, Aug. 2016.
4.
Chieko Ishifune and Koji Yasutomo : Notch-STAT5b signaling regulates the TCRalpha beta+ CD8alpha alpha+ intraepithelial lymphocytes in the small intestine, 17th International Congress of Mucosal Immunology, Berlin, Jul. 2015.
5.
Chieko Ishifune and Koji Yasutomo : Notch signaling is essential for the differentiation of CX3CR1 positive cells in lamina propria of the small intestine, 2012 International Symposium on Dendritic Cells., Daegu, Oct. 2012.
6.
Chieko Ishifune, Yoichi Maekawa and Koji Yasutomo : Functions of Notch signaling-dependent Thy1-expressing dendritic cells in immune response, The 94th AAI Annual Meeting, No.B-164, Miami, USA, May 2007.
Proceeding of Domestic Conference:
1.
Chieko Ishifune and Koji Yasutomo : Notch signal controls final differentiation of TCRγδ+CD8αα+ intraepithelial lymphocytes in the small intestine, 第51回 日本免疫学会学術集会, Dec. 2022.
2.
Chieko Ishifune and Koji Yasutomo : Notch-mediated final differantiation of TCRgd+CD8aa+ intraepithelial lymphocyte in the small intestine, 第47回 日本免疫学会学術集会, Dec. 2018.
3.
Chieko Ishifune, Shin-ichi Tsukumo and Koji Yasutomo : Notch signal controls the number of TCRab+CD8aa+ intraepithelial lymphocytes via phospholipid asymmetry by maintaining flippase ATP8a2, 第46回 日本免疫学会学術集会, Dec. 2017.
4.
Chieko Ishifune and Koji Yasutomo : Notchシグナルによる細胞膜リン脂質の極性制御を介した腸管上皮間リンパ球の制御, 第32回自己免疫研究会, Jul. 2017.
Chieko Ishifune, Yoichi Maekawa and Koji Yasutomo : Notch-Rbpj regulates Eat-Me singal of TCRab+CD8aa+ intraepithelial lymphocytes by controlling fllipase ATP8a2, 第45回日本免疫学会学術集会, Dec. 2016.
7.
N Doi, Chieko Ishifune, Koji Yasutomo, T Miura, Y Sakai, K Fujimoto, S Harada, K Yoshimura, Masako Nomaguchi and Akio Adachi : Replication and pathogenicity of HIV-1rmt: towards evaluation of viral growth ability in gut-derived cells, 第64回日本ウイルス学会学術集会, Oct. 2016.
8.
Chieko Ishifune, Yoichi Maekawa, Katsuto Hozumi and Koji Yasutomo : Notch-STAT5b is central for the differentiation of TCR alpha beta+ CD8 alpha alpha+ intraepithelial lymphocytes, 第44回日本免疫学会学術集会, Nov. 2015.
9.
Chieko Ishifune, Maekawa Yoichi, Katsuto Hozumi and Koji Yasutomo : Notch regulates the differentiation of TCRalpha beta+ CD8alpha alpha+ intraepithelial lymphocytes in the small intestine through Stat5b signaling, 第43回 日本免疫学会学術集会, Dec. 2014.
10.
Chieko Ishifune, Yoichi Maekawa, Katsuto Hozumi and Koji Yasutomo : Notch signaling regulates the differentiation of TCR alpha beta+ CD8alpha alpha+ Thy1- intraepithelial lymphocytes in the small intestine, 第42回日本免疫学会学術集会, Dec. 2013.
11.
Chieko Ishifune, Yoichi Maekawa and Koji Yasutomo : Notch signaling is essential for the differentiation of CX3CR1+ cell in lamina propria of the small intestine, 第41回日本免疫学会総会, Dec. 2012.
Chieko Ishifune, 前川 洋一 and 安友 康二 : NotchリガンドDll-1誘導性の樹状細胞による原虫感染克服の試み, 第75回日本寄生虫学会大会, May 2006.
Et cetera, Workshop:
1.
Chieko Ishifune and Koji Yasutomo : Notch signal controls number of TCRalphabeta+CD8alphaalpha+IEL thorough regulating phospholipid asymmetry, 6th joint meeting; Tokushima University, Gifu University, KAIST and Seoul National University, Sep. 2017.
2.
Chieko Ishifune and Koji Yasutomo : Notch and Intraepithelial Lymphocytes., 5th joint meeting; Tokushima University, KAIST and Seoul National University, Sep. 2016.
3.
Chieko Ishifune and Koji Yasutomo : Notch-STAT5b Signaling Regulates the TCRαβ+CD8αα+ Intraepithelial Lymphocytes in the Small Intestine., 4th joint meeting; Tokushima University, KAIST and Seoul National University, Oct. 2015.
4.
Chieko Ishifune and 安友 康二 : 難治性腸炎の発症に関わる腸管上皮細胞間リンパ球の分化制御, 第一回病因研究会別府シンポジウム, Jan. 2015.
5.
Chieko Ishifune and Koji Yasutomo : Control of intraepithelial lymphocytes by Notch signaling., 3rd joint meeting; Tokushima University, KAIST and Seoul National University, Sep. 2014.
6.
Chieko Ishifune and 安友 康二 : 自己免疫性大腸炎におけるNotchシグナルによる腸管上皮間T細胞の維持調節, 自己免疫研究会, Jul. 2014.
7.
Chieko Ishifune and Koji Yasutomo : Requirement of Notch signaling for the development of intestinal intraepithelial T lymphocytes, The Third Bizan Immunology Symposium at The University of Tokushima (BISUT3), "Immune System Development, Deviation, and Regulation", Feb. 2014.
8.
Chieko Ishifune and Koji Yasutomo : The differentiation of CX3CR1+CD11c+ cells by Notch signaling., 2nd joint meeting; Tokushima University, KAIST and Seoul National University, Sep. 2013.
9.
Chieko Ishifune and Koji Yasutomo : Notch-mediated regulation of intestinal CD11c+ cells, The Second Immunology Symposium of The University of Tokushima Immune Development, Deviation, and Regulation 25 January 2013, Jan. 2013.
10.
Chieko Ishifune and Koji Yasutomo : Notch-mediated regulation of intestinal antigen presenting cells., 1st joint meeting; Tokushima University, KAIST and Seoul National University, Dec. 2012.
Elucidation of the mechanism of Notch-mediated acquisition of interepithelial lymphocyte diversity in the small intestine using single-cell analysis technology (Project/Area Number: 23K06585 )
Elucidation of the regulation mechanisms of maturation and TCR repertoire of small intestinal interepithelial lymphocytes by Notch signaling (Project/Area Number: 18K15191 )
Elucidation of maintenance mechanism of intestinal homeostasis through control of small intestinal epithelial lymphocytes by Notch signal (Project/Area Number: 15K19131 )
Regulation of differentiation of the small intestinal lamina proprial CX3CR1-positive cells by the Notch signal. (Project/Area Number: 24890146 )