Hiroshi Watarai

Hiroshi Watarai
Kanazawa University | Kindai · Department of Immunology and Stem Cell Biology

Professor

About

147
Publications
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Publications

Publications (147)
Article
Full-text available
Author Summary T cells are a diverse group of immune cells involved in cell-mediated acquired immunity. One subset of T cells is the innate-like invariant natural killer T (iNKT) cells that recognize glycolipid ligands on target cells instead of peptides. We know that functionally distinct subtypes of iNKT cells are involved in specific pathologies...
Article
Full-text available
NKT cells demonstrate antitumor activity when activated to produce Th1 cytokines by DCs loaded with alpha-galactosylceramide, the prototypic NKT cell-activating glycolipid antigen. However, most patients do not have sufficient numbers of NKT cells to induce an effective immune response in this context, indicating a need for a source of NKT cells th...
Article
Full-text available
Airway hypersensitive reaction (AHR) is an animal model for asthma, which is caused or enhanced by environmental factors such as allergen exposure. However, the precise mechanisms that drive AHR remain unclear. We identified a novel subset of natural killer T (NKT) cells that expresses the interleukin 17 receptor B (IL-17RB) for IL-25 (also known a...
Article
Full-text available
Dendritic cells (DC) that are stimulated with inflammatory mediators can maturate and migrate from nonlymphoid tissues to lymphoid organs to initiate T cell-mediated immune responses. This migratory step is closely related to the maturation of the DC. In an attempt to identify chemokine receptors that might influence migration and are selectively e...
Article
This protocol describes methods to identify, purify and culture CD1d restricted invariant natural killer T (iNKT) cells from mouse tissue or human blood samples. The methods for identification and purification of iNKT cells are based on the interaction between iNKT cell receptor and its ligand. The iNKT cell receptor is composed of the invariant V...
Article
Background: Melioidosis is an infectious disease caused by Burkholderia pseudomallei. In infected mice, IFN-γ can provide protection against B. pseudomallei infection. Invariant Natural Killer T (iNKT) cells are a subpopulation of T lymphocytes, activated by recognition of glycolipid ligands such as α-Galactosylceramide presented by CD1d, produce...
Article
Full-text available
Marine sponges are an excellent source of biologically active secondary metabolites. We focus on deep-sea sponges for our discovery study. A marine sponge Cladocroce sp. exhibited cytotoxic activity in the bioactivity screening. From this sponge a previously unreported cytotoxic glycosphingolipid, calyxoside B, was isolated and the structure of thi...
Article
Full-text available
Objective Signal-transducing adaptor protein (STAP) family members function as adaptor molecules and are involved in several events during immune responses. Notably however, the biological functions of STAP-1 in other cells are not known. We aimed to investigate the functions of STAP-1 in invariant natural killer T (iNKT) cells and iNKT cell-depend...
Article
Full-text available
The advent of image-activated cell sorting and imaging-based cell picking has advanced our knowledge and exploitation of biological systems in the last decade. Unfortunately, they generally rely on fluorescent labeling for cellular phenotyping, an indirect measure of the molecular landscape in the cell, which has critical limitations. Here we demon...
Article
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Droplet microfluidics has become a powerful tool in precision medicine, green biotechnology, and cell therapy for single-cell analysis and selection by virtue of its ability to effectively confine cells. However, there remains a fundamental trade-off between droplet volume and sorting throughput, limiting the advantages of droplet microfluidics to...
Article
Thymic stromal lymphopoietin (TSLP) acts on dendritic cells (DCs), which prime helper T (Th) cells to become type 2 cytokine producing cells. Recently, a different set of populations of TSLP-responsive DCs has been discovered. Here, we identified two populations of CD103loEpCAMhi migratory DCs (fraction I and fraction II) that accumulated in skin-d...
Article
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By virtue of the combined merits of flow cytometry and fluorescence microscopy, imaging flow cytometry (IFC) has become an established tool for cell analysis in diverse biomedical fields such as cancer biology, microbiology, immunology, hematology, and stem cell biology. However, the performance and utility of IFC are severely limited by the fundam...
Preprint
Full-text available
By virtue of the combined merits of flow cytometry and fluorescence microscopy, imaging flow cytometry (IFC) has become an established tool for cell analysis in diverse biomedical fields such as cancer biology, microbiology, immunology, hematology, and stem cell biology. However, the performance and utility of IFC are severely limited by the fundam...
Article
α-Galactosylceramide (α-GalCer) is recognized by the CD1d proteins on antigen-presenting cells at the ceramide moiety and the galactose moiety is presented to iNKT cells, which stimulates the immune responses. However, the immune suppression by repeated injections of α-GalCer has discouraged its development as an anti-cancer agent. To overcome the...
Article
Background: Burkholderia thailandensis is a non-pathogenic bacterium that is closely related to B. pseudomallei. Invariant natural killer T (iNKT) cells are innate lymphoid cells that play a role in bacterial infections; however the iNKT cells in B. thailandensis infections are still uncharacterized. Objective: To study the cytokine production i...
Article
Full-text available
Zinc (Zn) is an essential nutrient and its deficiency causes immunodeficiency and skin disorders. Various cells including mast cells release Zn-containing granules when activated; however, the biological role of the released Zn is currently unclear. Here we report our findings that Zn transporter ZnT2 is required for the release of Zn from mast cel...
Article
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Primary tumours establish metastases by interfering with distinct organs. In pre-metastatic organs, a tumour-friendly microenvironment supports metastatic cells and is prepared by many factors including tissue resident cells, bone marrow-derived cells and abundant fibrinogen depositions. However, other components are unclear. Here, we show that a t...
Article
Full-text available
Invariant natural killer T (iNKT) cells are a unique T cell subset that exhibits characteristics of both innate immune cells and T cells. They express Vα14-Jα18 (Trav11-Traj18) as an invariant chain of the T cell receptor (TCR) and are restricted to the MHC class I-like monomorphic antigen presenting molecule CD1d. iNKT cells are known as immune re...
Article
Laser-scanning confocal fluorescence microscopy is an indispensable tool for biomedical research by virtue of its high spatial resolution. Its temporal resolution is equally important, but is still inadequate for many applications. Here we present a confocal fluorescence microscope that, for the first time to our knowledge, surpasses the highest po...
Article
Full-text available
iNKT cells play important roles in immune regulation by bridging the innate and acquired immune systems. The functions of iNKT cells have been investigated in mice lacking the Traj18 gene segment that were generated by traditional embryonic stem cell technology, but these animals contain a biased T cell receptor (TCR) repertoire that might affect i...
Article
Foxp3 controls the development and function of regulatory T (Treg) cells, but it remains elusive how Foxp3 functions in vivo. Here, we established mouse models harboring three unique missense Foxp3 mutations that were identified in patients with the autoimmune disease IPEX. The I363V and R397W mutations were loss-of-function mutations, causing mult...
Article
Full-text available
Interleukin-22 (IL-22) acts protectively and harmfully on intestinal tissue depending on the situation; therefore, IL-22 signaling needs to be tightly regulated. IL-22 binding protein (IL-22BP) binds IL-22 to inhibit IL-22 signaling. It is expressed in intestinal and lymphoid tissues, although its precise distribution and roles have remained unclea...
Article
Aside from a role in clot dissolution, the fibrinolytic factor, plasmin is implicated in tumorigenesis. Although abnormalities of coagulation and fibrinolysis have been reported in multiple myeloma patients, the biological roles of fibrinolytic factors in multiple myeloma (MM) using in vivo models have not been elucidated. In this study, we establi...
Conference Paper
Microalgae have been receiving great attention for their ability to produce biomaterials that are applicable for food supplements, drugs, biodegradable plastics, and biofuels. Among such microalgae, Euglena gracilis has become a popular species by virtue of its capability of accumulating useful metabolites including paramylon and lipids. In order t...
Chapter
Invariant natural killer T (iNKT) cells are a subpopulation of T lymphocytes with unique specificities against (glyco)lipids presented in the context of CD1d belonging to major histocompatibility complex (MHC) class Ib. They recognise microbially encoded or synthetic glycolipids directly through their CD1d-restricted T-cell receptors (TCRs) other t...
Article
A specialized bone marrow microenvironment (niche) regulates hematopoietic stem cell (HSC) self- renewal and commitment. For successful donor-HSC engraftment, the niche must be emptied via myeloablative irradiation or chemotherapy. However, myeloablation can cause severe complications and even mortality. Here, we report that the essential amino aci...
Article
Full-text available
Zinc transporters play a critical role in spatiotemporal regulation of zinc homeostasis. Although disruption of zinc homeostasis has been implicated in disorders such as intestinal inflammation and aberrant epithelial morphology, it is largely unknown which zinc transporters are responsible for the intestinal epithelial homeostasis. Here, we show t...
Data
Effect of supplementation of Zn on Zip7 deletion-induced upregulation of ER stress-related gene expression. (A) Quantitative PCR analysis of Derl3, CHOP, and Creld2 in cultured organoids from Zip7Cont and Zip7ΔIEC mice 48 h after 4-OHT treatment with or without ZnSO4 (1 μM). (B) Quantitative PCR analysis of Derl3 and CHOP Zip7-/- MEF cells treated...
Data
In situ hybridization analysis of Zip7 in small intestine. In situ hybridization of Zip7 in small intestine from C57BL/6J mice. Scale bar: 100 μm. (TIF)
Data
Primer sequences of mouse genes used for quantitative real-time PCR. (PDF)
Data
Generation of Zip7flox/flox mice. (A) Targeting vectors with three LoxP insertions. In Zip7flox mice, two LoxP sites were inserted into introns 4 and 6, respectively. Zip7Δflox mice were obtained by crossing with Cre-transgenic mice. Gray triangles indicate LoxP sites; closed boxes indicate exons. (B) Zip7 deletion. MEF cells derived from Zip7-/- m...
Data
Schematic diagram for intestinal organoid culture. Intestinal crypts isolated from Zip7Cont and Zip7ΔIEC mice were embedded in Matrigel and incubated with 1 μM 4-OHT. (TIF)
Data
Zip7 in Lgr5+ stem cells is required for regeneration after irradiation. (A) Representative H&E staining of intestines from Zip7Cont and Zip7ΔLgr5 mice. (B) Organoids established from Zip7ΔLgr5 mice with (lower panel) or without (upper panel) 4-OHT treatment for 48 h. (C) The tamoxifen (TM) induced-induced Zip7 deletion and radiation ragimen used i...
Data
ZIP7 governs organoid self-renewal independent of Paneth cells. (A) In vitro Zip7-deletion protocol to examine the effect of added Wnt3a on intestinal organoids from Zip7Cont and Zip7ΔIEC mice. (B) Crypts cultured in standard medium plus Wnt3a. Crypts from a control mouse showed rounded cysts. Exogenous Wnt3a did not support the growth of crypts fr...
Data
Zip7 deletion-induced gene expression changes. Quantitative PCR analysis of Xbp1 (A) and Mt1 (B) mRNA in cultured organoids from Zip7Cont and Zip7ΔIEC mice 48 h after 4-OHT treatment. (TIF)
Data
RNA expression profiles of sorted Lgr5hi and Lgr5int cells. DNA microarray expression experiments were performed with sorted Lgr5hi and Lgr5int cells. (A) Heat map of stem cell-related genes in sorted Lgr5hi and Lgr5int cells from Zip7Cont and Zip7ΔIEC mice, showing a clear separation between the different Lgr5 populations. (B) Heat map of 1.8-fold...
Data
RNA expression profiles of MEF cells from Zip7+/- and Zip7-/- mice. Heat map of 2.0-fold change of gene epxression between Zip7+/- and Zip7-/- MEF cells. Columns represent different cell types, and rows represent differentially expressed genes. (TIF)
Article
Experimental autoimmune uveoretinitis (EAU) represents an experimental model for human endogenous uveitis, which is caused by Th1/Th17 cell-mediated inflammation. Natural killer T (NKT) cells recognize lipid antigens and produce large amounts of cytokines upon activation. To examine the role of NKT cells in the development of uveitis, EAU was elici...
Article
Understanding metabolism in live microalgae is crucial for efficient biomaterial engineering, but conventional methods fail to evaluate heterogeneous populations of motile microalgae due to the labelling requirements and limited imaging speeds. Here, we demonstrate label-free video-rate metabolite imaging of live Euglena gracilis and statistical an...
Article
Conventional dendritic cells (cDCs) present α-galactosylceramide (αGC) to invariant natural killer T (iNKT) cells through CD1d. Among cDC subsets, CD8(+) DCs efficiently induce IFN-γ production in iNKT cells. Using fluorescence-labeled αGC, we showed that CD8(+) DCs incorporated larger amounts of αGC and kept it intact longer than CD8(-) DCs. Histo...
Article
Full-text available
The interleukin-17 (IL-17) family of cytokines has emerged as a critical player in inflammatory diseases. Among them, IL-25 has been shown to be important in allergic inflammation and protection against parasitic infection. Here we have demonstrated that IL-17B, a poorly understood cytokine, functions to inhibit IL-25-driven inflammation. IL-17B an...
Patent
Full-text available
Provided are an iPS cell derived from a somatic cell such as an NKT cell, having the α-chain region of the T cell antigen receptor gene rearranged to uniform Vα-Jα in an NKT cell receptor-specific way, NKT cells differentiated from the iPS cell, a method of creating the same, and an immune cell therapy agent prepared using cells differentiated from...
Patent
Full-text available
A compound represented by the following formula (1): wherein R1 is an aldopyranose residue wherein the 6-hydroxyl group is optionally alkylated, R2 is a C1-26 hydrocarbon group optionally having substituent(s), R3 is a hydrogen atom or a C1-26 hydrocarbon group optionally having substituent(s), R4 is a C1-21 hydrocarbon group optionally having sub...
Article
Immunological memory has been regarded as a unique feature of the adaptive immune response mediated in an antigen-specific manner by T and B lymphocytes. However, natural killer (NK) cells and γδT cells, which traditionally are classified as innate immune cells, have been shown in recent studies to have hallmark features of memory cells. Invariant...
Article
Full-text available
NKT cells are characterized by their expression of an NKT-cell-specific invariant antigen-receptor α chain encoded by Vα14Jα18 gene segments. These NKT cells bridge the innate and acquired immune systems to mediate effective and augmented responses; however, the limited number of NKT cells in vivo hampers their analysis. Here, two lines of induced...
Article
Contact hypersensitivity (CHS) has been widely used to study cutaneous immune responses, as a prototype of delayed-type hypersensitivity. Although natural killer T (NKT) cells have been assumed to play an important role in CHS, their role is controversial. Here, we report the role of NKT cells in the sensitization phase of CHS, by promoting the sur...
Article
RCAI-147 is one of the hydroxylated analogues of KRN7000 which is known as a ligand for the activation of CD1d mediated invariant natural killer T cells (iNKT cells) and releases both T helper 1 (Th1) cytokines such as IFN-γ and T helper 2 (Th2) cytokines such as IL-4. KRN7000 has been anticipated as an antitumor drug or an adjuvant for viral infec...
Article
RCAI-80 is one of the ester analogs of KRN7000 (α-galactosylceramide). This compound released mainly T helper 2 (Th2) cytokines, such as IL-4 rather than T helper 1 (Th1) cytokines, such as IFNγ from the invariant natural killer T (iNKT) cells. In addition, it has been known that some of the hydroxylated derivatives of KRN7000 make the cytokine sec...
Patent
Full-text available
The invention provides esterified α-galactosylceramides effective for cancer treatment and the like, and a medicament containing same. In particular, the invention relates to a compound represented by the formula (I): wherein R1 is a hydrocarbon group having a carbon number of 1 to 30, R2 is a hydrocarbon group having a carbon number of 1 to 20, R...
Patent
Full-text available
The invention provides a glycolipid effective for cancer treatment and the like and a synthetic intermediate therefor, as well as a medicament containing the glycolipid and the like. The glycolipid is represented by the formula (1) or a salt thereof
Article
Full-text available
Background: The commensal yeast Candida albicans is a major cause of invasive fungal infections. Despite treatment with antifungal agents, the mortality rate attributed to these types of infection is high. Although numerous cases have been reported regarding a poor outcome for patients with bacterial and C. albicans coinfection, the mechanisms by...
Article
Macrophages play a critical role in chronic inflammation and metabolic diseases. We identified a longer splice variant of ubiquitin specific protease (USP) 2-69 as a novel molecule that modulates pathways implicated in metabolic disorders. Expression levels of aP2/FABP4 and PAI-1/SERPINE1 genes were increased by 4- and 1.8-fold, respectively, after...
Article
Pancreatic islet transplantation is an attractive therapy for the treatment of insulin-dependent diabetes mellitus. However, the low efficiency of this procedure necessitating sequential transplantations of islets with the use of 2-3 donors for a single recipient, mainly due to the early loss of transplanted islets, hampers its clinical application...
Article
Full-text available
We synthesized seven new analogues of KRN7000: RCAI-133 and 154–159. RCAI-154, 156 and 158 are secondary-amide analogues having a hydroxy, a methyl or two methyl groups at the α-position of a linear C18-acyl chain, respectively. RCAI-155, 157 and 159 are corresponding N-methylated tertiary amide analogues, and RCAI-133 is the N-methylated KRN7000....
Article
KRN7000 is one of the α-galactosylceramides, which has a 2-hexacosanoylamino-3,4-dihydroxyoctadecyl group. This compound, known as a ligand for the activation of CD1d mediated invariant natural killer T cells (iNKT cells) which release both T helper 1 (Th1) cytokines such as IFNγ and Th2 cytokines such as IL-4, has been anticipated as an antitumor...
Article
We synthesized ten new analogs of 6'-modified KRN7000 (A): RCAI-58, 61, 64, 83, 85-87, 113, 119, and 125. They could be synthesized by α-selective galactosylation of ceramide 9 with the 6-modified d-galactopyranosyl fluorides (8a-8f) or l-arabinopyranosyl fluoride (17), or by etherification of the known alcohol 19. Bioassay of the ten analogs demon...
Article
Candida albicans is a major cause of invasive fungal infections. Mortality attributable to candidemia is very high, even when patients are treated with adequate antifungal agents. Therefore, it is important to investigate the mechanisms of immune response to C. albicans. Invariant natural killer T (iNKT) cells, innate lymphocytes that express an in...
Chapter
Natural killer T (NKT) cells are characterized by the expression of an invariant Vα14Jα18 receptor that recognizes glycolipids such as α-galactosylceramide (α-GalCer) in conjunction with CD1d molecule. Functionally, NKT cells can act as innate immune cells but can also bridge the innate and acquired immune systems. A prime example of one such funct...
Article
RCAI-84, 91, and 105-108 (1-6), the analogs of KRN7000 (A) with a ureido or a thioureido linkage instead of a carboxamido bond, were synthesized to examine their immunostimulatory activity against mouse lymphocytes. According to their bioassay, the ureido analog of KRN7000 [RCAI-105 (1)] and its 6'-O-methylated derivative [RCAI-106 (4)] induced a l...