M Itakura, M Moritani, F Tashiro, C Hashimoto, J Miyazaki, S Li, E Kudo, H Iwahana, Yoshio Hayashi, T Sano and K Yoshimoto : Diabetes, --- Pancreatic IL-10 accelerates autoimmune insulitis and diabetes in transgenic NOD mice ---, Elsevier Science, 1994.
21.
Yoshio Hayashi, N Haneji and H Hamano : Transfer of murine Sjogren-like autoimmune lesions to SCID mice and its prevention by anti-CD4 and -TCRV β8 treatment, State of Art, 1994.
Mitsunobu Sato, Yoshio Hayashi and Hideo Yoshida : Immunopharmacological aspects of OK-432 in humans, --- Effect of immuno-therapy with streptococcal preparation,OK-432, on the peripheral killer lymphocyte population in patients with head and neck cancer ---, 1986.
25.
Mitsunobu Sato, Yoshio Hayashi and Hideo Yoshida : Recent Advances in Chemotherapy, --- Effect of postoperative immunotherapy with streptpcoccal preparation OK-432 on different immunologic parameters in patients with head and neck cancer ---, 1985.
学術論文(審査論文):
1.
Akiko Yamada, Aya Ushio, Rieko Arakaki, Takaaki Tsunematsu, Yasusei Kudo, Yoshio Hayashi and Naozumi Ishimaru : Impaired expansion of regulatory T cells in a neonatal thymectomy-induced autoimmune mouse model., The American Journal of Pathology, Vol.185, No.11, 2886-2897, 2015.
(要約)
Neonatal thymectomy in certain mouse strains is known to induce organ-specific autoimmunity due to impaired functions of T cells, including Foxp3(+) regulatory T (Treg) cells in the thymus. The precise mechanism underlying the induction of autoimmunity by neonatal thymectomy remains unclear. One possibility is that depletion of Treg cells breaks down peripheral tolerance. We examined the functions of Treg cells by using a murine Sjögren syndrome model of NFS/sld mice that underwent neonatal thymectomy. The ratio of Treg cells to effector memory phenotype T cells in thymectomy mice was significantly lower than that of nonthymectomy mice. In addition, in vitro induction of peripherally induced Treg cells by transforming growth factor-β (TGF-β) using naive T cells from Sjögren syndrome model mice was severely impaired. The mRNA expression of TGF-β receptor I and II and Smad3 and -4 in the TGF-β-induced signal transduction pathway of Treg cells in this Sjögren syndrome model were lower than those of control mice. In addition, Treg cells in this Sjögren syndrome model exhibited an interferon-γ-producing Th1-like phenotype that resembled effector T cells. In conclusion, these results suggest that abnormal expansion and differentiation of Treg cells and inflammatory cytokines produced by Treg cells contribute to the development of autoimmunity.
Akihiko Iwasa, Rieko Arakaki, Naoko Honma, Aya Ushio, Akiko Yamada, Tomoyuki Kondo, Emi Kurosawa, Satoko Kujiraoka, Takaaki Tsunematsu, Yasusei Kudo, Eiji Tanaka, Noriko Yoshimura, Nobuhiro Harada, Yoshio Hayashi and Naozumi Ishimaru : Aromatase controls Sjögren syndrome-like lesions through monocyte chemotactic protein-1 in target organ and adipose tissue-associated macrophages., The American Journal of Pathology, Vol.185, No.1, 151-161, 2015.
(要約)
Several autoimmune diseases are known to develop in postmenopausal women. However, the mechanism by which estrogen deficiency influences autoimmunity is unknown. Aromatase is an enzyme that converts androgens to estrogens. Herein, we used female aromatase gene knockout (ArKO) mice as a model of estrogen deficiency to investigate the molecular mechanism that underlies the onset and development of autoimmunity. Histological analyses showed that inflammatory lesions in the lacrimal and salivary glands of ArKO mice increased with age. Adoptive transfer of spleen cells or bone marrow cells from ArKO mice into recombination activating gene 2 knockout mice failed to induce the autoimmune lesions. Expression of mRNA encoding proinflammatory cytokines and monocyte chemotactic protein-1 increased in white adipose tissue of ArKO mice and was significantly higher than that in wild-type mice. Moreover, an increased number of inflammatory M1 macrophages was observed in white adipose tissue of ArKO mice. A significantly increased monocyte chemotactic protein-1 mRNA expression of the salivary gland tissue in ArKO was found together with adiposity. Furthermore, the autoimmune lesions in a murine model of Sjögren syndrome were exacerbated by administration of an aromatase inhibitor. These results suggest that aromatase may play a key role in the pathogenesis of Sjögren syndrome-like lesions by controlling the target organ and adipose tissue-associated macrophage.
Rieko Arakaki, Hiroshi Eguchi, Akiko Yamada, Yasusei Kudo, Akihiko Iwasa, Enkhmaa Tserennadmid, Hotta Fumika, Mitamura-Aizawa Sayaka, Yoshio Hayashi and Naozumi Ishimaru : Anti-Inflammatory Effects of Rebamipide Eyedrop Administration on Ocular Lesions in a Murine Model of Primary Sjögren's Syndrome., PLoS ONE, Vol.9, No.5, e98390, 2014.
(要約)
Topical therapy is effective for dry eye, and its prolonged effects should help in maintaining the quality of life of patients with dry eye. We previously reported that the oral administration of rebamipide (Reb), a mucosal protective agent, had a potent therapeutic effect on autoimmune lesions in a murine model of Sjögren's syndrome (SS). However, the effects of topical treatment with Reb eyedrops on the ocular lesions in the murine model of SS are unknown. Reb eyedrops were administered to the murine model of SS aged 4-8 weeks four times daily. Inflammatory lesions of the extraorbital and intraorbital lacrimal glands and Harderian gland tissues were histologically evaluated. The direct effects of Reb on the lacrimal glands were analyzed using cultured lacrimal gland cells. Tear secretions of Reb-treated mice were significantly increased compared with those of untreated mice. In addition to the therapeutic effect of Reb treatment on keratoconjunctivitis, severe inflammatory lesions of intraorbital lacrimal gland tissues in this model of SS were resolved. The mRNA expression levels of IL-10 and mucin 5Ac in conjunctival tissues from Reb-treated mice was significantly increased compared with those of control mice. Moreover, lactoferrin production from lacrimal gland cells was restored by Reb treatment. Topical Reb administration had an anti-inflammatory effect on the ocular autoimmune lesions in the murine model of SS and a protective effect on the ocular surfaces.
Ashrin Nur Meinar, Rieko Arakaki, Akiko Yamada, Tomoyuki Kondo, Mie Kurosawa, Yasusei Kudo, Megumi Watanabe, Tetsuo Ichikawa, Yoshio Hayashi and Naozumi Ishimaru : A critical role for thymic stromal lymphopoietin in nickel-induced allergy in mice., The Journal of Immunology, Vol.192, No.9, 4025-4031, 2014.
(要約)
Ni is the most frequent cause of contact allergy induced by metals. However, the underlying mechanism of this induction is unknown. Our previous research demonstrates that activation of dendritic cells (DCs) through p38MAPK/MKK6 is required for Ni-induced allergy in mice. In the current study, we investigated the cellular and molecular mechanisms underlying Ni-induced allergy using a mouse model that involves injecting Ni into the ear, with or without Freund's incomplete or complete adjuvants. Nickel had greater potential to cause allergic reactions compared with palladium and gold. Among the proteins expressed at higher levels in mice with Ni-induced allergy, we focused on thymic stromal lymphopoietin (TSLP), which is produced in abundance by keratinocytes. We detected increased expression of the TSLP receptor (TSLPR) in DCs from cervical lymph nodes of mice with Ni-induced allergy, suggesting that DCs in ear tissues were activated through TSLPR signaling induced by keratinocyte-derived TSLP. Furthermore, delayed-type hypersensitivity reactions in mice with Ni-induced allergy were decreased significantly by injection of a Tslp-short interfering RNA along with atelocollagen in the ear skin. These results suggest that Ni allergy may be triggered by a TSLP/TSLPR-mediated interaction between epithelial and immune cells.
Yosuke Shikama, Naozumi Ishimaru, Yasusei Kudo, Yukiko Bandou, N Aki, Yoshio Hayashi and Makoto Funaki : Effects of Free Fatty Acids on Human Salivary Gland Epithelial Cells., Journal of Dental Research, Vol.92, No.6, 540-546, 2013.
(要約)
Obesity and type 2 diabetes (T2D) are characterized by decreased insulin sensitivity and higher concentrations of free fatty acids (FFAs) in plasma. Among FFAs, saturated fatty acids (SFAs), such as palmitate, have been proposed to promote inflammatory responses. Primary Sjögren's syndrome (SS) is an autoimmune disease characterized by inflammatory mononuclear cell infiltration and destruction of epithelial cells in the salivary and lacrimal glands. IL-6 production and -fodrin degradation are increased in salivary gland epithelial cells of patients with primary SS. Although previous studies have shown a link between SS and either dyslipidemia or T2D, little is known about the clinical significance of FFAs in primary SS. Here we report that SFAs, but not unsaturated fatty acids, induced IL-6 production via NF-B and p38 MAPK activation in human salivary gland epithelial cells. Moreover, palmitate induced apoptosis and -fodrin degradation by caspase-3 activation. Unlike salivary gland epithelial cells, induction of IL-6 production and the degradation of -fodrin in response to palmitate were undetectable in squamous carcinoma cells and keratinocytes. Taken together, SFAs induced IL-6 production and -fodrin degradation in salivary gland epithelial cells, implicating a potential link between the pathogenesis of primary SS and SFAs level in plasma.
Ritsuko 徳永 律子 Oura, Rieko Arakaki, Akiko Yamada, Yasusei Kudo, Eiji Tanaka, Yoshio Hayashi and Naozumi Ishimaru : Induction of Rapid T Cell Death and Phagocytic Activity by Fas-Deficient lpr Macrophages., The Journal of Immunology, 2013.
(要約)
Peripheral T cells are maintained by the apoptosis of activated T cells through the Fas-Fas ligand system. Although it is well known that normal T cells fail to survive in the Fas-deficient immune condition, the molecular mechanism for the phenomenon has yet to be elucidated. In this study, we demonstrate that rapid cell death and clearance of normal T cells were induced by Fas-deficient lpr macrophages. Transfer of normal T cells into lpr mice revealed that Fas expression on donor T cells was promptly enhanced through the IFN-/IFN-R. In addition, Fas ligand expression and phagocytic activity of lpr macrophages were promoted through increased NF-B activation. Controlling Fas expression on macrophages plays an essential role in maintaining T cell homeostasis in the peripheral immune system. Our data suggest a critical implication to the therapeutic strategies such as transplantation and immunotherapy for immune disorder or autoimmunity related to abnormal Fas expression.
BACKGROUND: Although autoimmunity in MRL/lpr mice occurs due to a defect in Fas-mediated cell death of T cells, the role of Fas-independent apoptosis in pathogenesis has rarely been investigated. We have recently reported that receptor activator of nuclear factor (NF)-B ligand (RANKL)-activated dendritic cells (DCs) play a key role in the pathogenesis of rheumatoid arthritis (RA) in MRL/lpr mice. We here attempted to establish a new therapeutic strategy with RANKL-activated DCs in RA by controlling apoptosis of peripheral T cells. Repeated transfer of RANKL-activated DCs into MRL/lpr mice was tested to determine whether this had a therapeutic effect on autoimmunity. METHODS AND FINDING: Cellular and molecular mechanisms of Fas-independent apoptosis of T cells induced by the DCs were investigated by in vitro and in vivo analyses. We demonstrated that repeated transfers of RANKL-activated DCs into MRL/lpr mice resulted in therapeutic effects on RA lesions and lymphoproliferation due to declines of CD4(+) T, B, and CD4(-)CD8(-) double negative (DN) T cells. We also found that the Fas-independent T-cell apoptosis was induced by a direct interaction between tumor necrosis factor (TNF)-related apoptosis-inducing ligand-receptor 2 (TRAIL-R2) on T cells and TRAIL on Fas-deficient DCs in MRL/lpr mice. CONCLUSION: These results strongly suggest that a novel Fas-independent apoptosis pathway in T cells maintains peripheral tolerance and thus controls autoimmunity in MRL/lpr mice.
Naozumi Ishimaru, Akiko Yamada, Takeshi Nitta, Rieko Arakaki, Martin Lipp, Yousuke Takahama and Yoshio Hayashi : CCR7 with S1P1 signaling through AP-1 for migration of Foxp3+ regulatory T-cells controls autoimmune exocrinopathy., The American Journal of Pathology, Vol.180, No.1, 199-208, 2012.
(要約)
Forkhead box p3-positive (Foxp3(+)) regulatory T cells (T(reg) cells) participate in maintaining peripheral immune tolerance and suppressing autoimmunity. We recently reported that in situ patrolling by C-C-chemokine receptor 7 (CCR7)(+) T(reg) cells in target organs is essential for controlling autoimmune lesions in Sjögren's syndrome. In the present study, the molecular mechanism underlying CCR7-mediated T(reg) cell migration was investigated in a mouse model. The impaired migratory response of Ccr7(-/-) T(reg) cells to sphingosine 1-phosphate (S1P) occurred because of defective association of S1P receptor 1 (S1P(1)) with a G coupled-protein. In addition, T-cell receptor (TCR)- and S1P(1)-mediated Ras-related C3 botulinum toxin substrate 1 (Rac-1), extracellular signal-related kinase (ERK), and c-Jun phosphorylation required for activator protein 1 (AP-1) transcriptional activity were significantly impaired in Ccr7(-/-) T(reg) cells. Surprisingly, the abnormal nuclear localization of Foxp3 was detected after abrogation of the c-Jun and Foxp3 interaction in the nucleus of Ccr7(-/-) T(reg) cells. These results indicate that CCR7 essentially controls the migratory function of T(reg) cells through S1P(1)-mediated AP-1 signaling, which is regulated through its interaction with Foxp3 in the nucleus.
Megumi Watanabe, Naozumi Ishimaru, Meinar Nur Ashrin, Rieko Arakaki, Akiko Yamada, Tetsuo Ichikawa and Yoshio Hayashi : A Novel DC Therapy with Manipulation of MKK6 Gene on Nickel Allergy in Mice, PLoS ONE, Vol.6, No.4, E19017, 2011.
(要約)
BACKGROUND: Although the activation of dermal dendritic cells (DCs) or Langerhans cells (LCs) via p38 mitogen-activated protein kinase (MAPK) plays a crucial role in the pathogenesis of metal allergy, the in vivo molecular mechanisms have not been identified and a possible therapeutic strategy using the control of dermal DCs or LCs has not been established. In this study, we focused on dermal DCs to define the in vivo mechanisms of metal allergy pathogenesis in a mouse nickel (Ni) allergy model. The effects of DC therapy on Ni allergic responses were also investigated. METHODS AND FINDING: The activation of dermal DCs via p38 MAPK triggered a T cell-mediated allergic immune response in this model. In the MAPK signaling cascade in DCs, Ni potently phosphorylated MAP kinase kinase 6 (MKK6) following increased DC activation. Ni-stimulated DCs could prime T cell activation to induce Ni allergy. Interestingly, when MKK6 gene-transfected DCs were transferred into the model mice, a more pronounced allergic reaction was observed. In addition, injection of short interfering (si) RNA targeting the MKK6 gene protected against a hypersensitivity reaction after Ni immunization. The cooperative action between T cell activation and MKK6-mediated DC activation by Ni played an important role in the development of Ni allergy. CONCLUSIONS: DC activation by Ni played an important role in the development of Ni allergy. Manipulating the MKK6 gene in DCs may be a good therapeutic strategy for dermal Ni allergy.
Yu Lei, Adiratna Mat Ripen, Naozumi Ishimaru, Izumi Ohigashi, Takashi Nagasawa, Lukas T. Jeker, Michael R. Bösl, Georg A. Holländer, Yoshio Hayashi, Rene Waal de Malefyt, Takeshi Nitta and Yousuke Takahama : Aire-dependent production of XCL1 mediates medullary accumulation of thymic dendritic cells and contributes to regulatory T cell development, The Journal of Experimental Medicine, Vol.208, No.2, 383-394, 2011.
(要約)
Dendritic cells (DCs) in the thymus (tDCs) are predominantly accumulated in the medulla and contribute to the establishment of self-tolerance. However, how the medullary accumulation of tDCs is regulated and involved in self-tolerance is unclear. We show that the chemokine receptor XCR1 is expressed by tDCs, whereas medullary thymic epithelial cells (mTECs) express the ligand XCL1. XCL1-deficient mice are defective in the medullary accumulation of tDCs and the thymic generation of naturally occurring regulatory T cells (nT reg cells). Thymocytes from XCL1-deficient mice elicit dacryoadenitis in nude mice. mTEC expression of XCL1, tDC medullary accumulation, and nT reg cell generation are diminished in Aire-deficient mice. These results indicate that the XCL1-mediated medullary accumulation of tDCs contributes to nT reg cell development and is regulated by Aire.
Small interfering RNA (siRNA)-mediated silencing of gene expression is rapidly becoming a powerful tool for molecular therapy. However, the rapid degradation of siRNAs and their limited duration of activity require efficient delivery methods. Atelocollagen (ATCOL)-mediated administration of siRNAs is a promising approach to disease treatment, including muscular atrophy. Herein, we report that ATCOL-mediated systemic administration of a myostatin-targeting siRNA into a caveolin-3-deficient mouse model of limb-girdle muscular dystrophy 1C (LGMD1C) induced a marked increase in muscle mass and a significant recovery of contractile force. These results provide evidence that ATCOL-mediated systemic administration of siRNAs may be a powerful therapeutic tool for disease treatment, including muscular atrophy.
Taro Adachi, Emi Kawakami, Naozumi Ishimaru, Takahiro Ochiya, Yoshio Hayashi, Hideyo Ohuchi, Masao Tanihara, Eiji Tanaka and Sumihare Noji : Delivery of small interfering RNA with a synthetic collagen poly(Pro-Hyp-Gly) for gene silencing in vitro and in vivo., Development Growth & Differentiation, Vol.52, No.8, 693-699, 2010.
(要約)
Silencing gene expression by small interfering RNAs (siRNAs) has become a powerful tool for the genetic analysis of many animals. However, the rapid degradation of siRNA and the limited duration of its action in vivo have called for an efficient delivery technology. Here, we describe that siRNA complexed with a synthetic collagen poly(Pro-Hyp-Gly) (SYCOL) is resistant to nucleases and is efficiently transferred into cells in vitro and in vivo, thereby allowing long-term gene silencing in vivo. We found that the SYCOL-mediated local application of siRNA targeting myostatin, coding a negative regulator of skeletal muscle growth, in mouse skeletal muscles, caused a marked increase in the muscle mass within a few weeks after application. Furthermore, in vivo administration of an anti-luciferase siRNA/SYCOL complex partially reduced luciferase expression in xenografted tumors in vivo. These results indicate a SYCOL-based non-viral delivery method could be a reliable simple approach to knockdown gene expression by RNAi in vivo as well as in vitro.
(キーワード)
Animals / Base Sequence / Cell Line, Tumor / Collagen / DNA Primers / Gene Silencing / Mice / RNA, Small Interfering / Reverse Transcriptase Polymerase Chain Reaction
Takashi Sakai, Li Liu, Xichuan Teng, Naozumi Ishimaru, Rika Mukai-Sakai, HoangNam Tran, Sun Mi Kim, Nobuya Sano, Yoshio Hayashi, Ryuji Kaji and Kiyoshi Fukui : Inflammatory disease and cancer with a decrease in Kupffer cell numbers in Nucling-knockout mice., International Journal of Cancer, Vol.126, No.5, 1079-1094, 2010.
(要約)
Nucling is a stress-inducible protein associated with apoptosomes. The cytochrome c-triggered formation of apoptosomes represents a key-initiating event in apoptosis. We have recently reported that Nucling regulates the apoptotic pathway by controlling the activation of NF-kappaB as well. Here we show that hepatocellular carcinoma (HCC) arising spontaneously against a background of hepatitis occurred more frequently in Nucling-knockout (KO) mice than wild-type (WT) mice. Biochemical serum testing revealed potential liver dysfunction with hypercholesterolemia in Nucling-KO males. In the background of Nucling-KO mice, we observed the up-regulation of TNFalpha, spontaneous NF-kappaB-activation and the induction of galectin-3 expression in liver. In addition, we observed a decrease in the number of Kupffer cells (KCs) in the KO mice. KCs are important for the hepatic immune system, acting as phagocytes or antigen-presenting cells (APCs). We found that KCs in Nucling-KO mice were apoptotic possibly through the up-regulation of TNFalpha. These observations indicate that Nucling is important for the regulation of NF-kappaB signals in liver. We propose that Nucling deficiency could be a powerful tool to reveal the NF-kappaB-related molecular networks leading to hepatitis and HCC development.
Naozumi Ishimaru, Takeshi Nitta, Rieko Arakaki, Akiko Yamada, Martin Lipp, Yousuke Takahama and Yoshio Hayashi : In situ Patrolling of Regulatory T cells is Essential for Protecting Autoimmune Exocrinopathy, PLoS ONE, Vol.5, No.1, e8588, 2010.
(要約)
Migration of T cells, including regulatory T (Treg) cells, into the secondary lymph organs is critically controlled by chemokines and adhesion molecules. However, the mechanisms by which Treg cells regulate organ-specific autoimmunity via these molecules remain unclear. Although we previously reported autoimmune exocrinopathy resembling Sjögren's syndrome (SS) in the lacrimal and salivary glands from C-C chemokine receptor 7 (CCR7)-deficient mice, it is still unclear whether CCR7 signaling might specifically affect the dynamics and functions of Treg cells in vivo. We therefore investigated the cellular mechanism for suppressive function of Treg cells via CCR7 in autoimmunity using mouse models and human samples. Patrolling Treg cells were detected in the exocrine organs such as lacrimal and salivary glands from normal mice that tend to be targets for autoimmunity while the Treg cells were almost undetectable in the exocrine glands of CCR7(-/-) mice. In addition, we found the significantly increased retention of CD4(+)CD25(+)Foxp3(+) Treg cells in the lymph nodes of CCR7(-/-) mice with aging. Although Treg cell egress requires sphingosine 1-phosphate (S1P), chemotactic function to S1P of CCR7-/- Treg cells was impaired compared with that of WT Treg cells. Moreover, the in vivo suppression activity was remarkably diminished in CCR7(-/-) Treg cells in the model where Treg cells were co-transferred with CCR7(-/-) CD25(-)CD4(+) T cells into Rag2(-/-) mice. Finally, confocal analysis showed that CCR7(+)Treg cells were detectable in normal salivary glands while the number of CCR7(+)Treg cells was extremely decreased in the tissues from patients with Sjögren's syndrome. These results indicate that CCR7 essentially governs the patrolling functions of Treg cells by controlling the traffic to the exocrine organs for protecting autoimmunity. Characterization of this cellular mechanism could have clinical implications by supporting development of new diagnosis or treatments for the organ-specific autoimmune diseases such as Sjögren's syndrome and clarifying how the local immune system regulates autoimmunity.
Akiko Yamada, Naozumi Ishimaru, Rieko Arakaki, Nobuhiko Katsunuma and Yoshio Hayashi : Cathepsin L inhivition prevents murine autoimmune diabetes via suppression of CD8+ T cell activity., PLoS ONE, Vol.5, No.9, e12894, 2010.
(要約)
Type 1 diabetes (T1D) is an autoimmune disease resulting from defects in central and peripheral tolerance and characterized by T cell-mediated destruction of islet β cells. To determine whether specific lysosomal proteases might influence the outcome of a T cell-mediated autoimmune response, we examined the functional significance of cathepsin inhibition on autoimmune T1D-prone non-obese diabetic (NOD) mice. Here it was found that specific inhibition of cathepsin L affords strong protection from cyclophosphamide (CY)-induced insulitis and diabetes of NOD mice at the advanced stage of CD8(+) T cell infiltration via inhibiting granzyme activity. It was discovered that cathepsin L inhibition prevents cytotoxic activity of CD8(+) T cells in the pancreatic islets through controlling dipeptidyl peptidase I activity. Moreover, the gene targeting for cathepsin L with application of in vivo siRNA administration successfully prevented CY-induced diabetes of NOD mice. Finally, cathepsin L mRNA expression of peripheral CD8(+) T cells from NOD mice developing spontaneous T1D was significantly increased compared with that from control mice. Our results identified a novel function of cathepsin L as an enzyme whose activity is essential for the progression of CD8(+) T cell-mediated autoimmune diabetes, and inhibition of cathepsin L as a powerful therapeutic strategy for autoimmune diabetes.
Yoshio Hayashi, Rieko Arakaki and Naozumi Ishimaru : Salivary gland and autoimmunity, The Journal of Medical Investigation : JMI, Vol.56, 185-191, 2009.
(要約)
Recent evidences suggest that the apoptotic pathway plays a central role in tolerazing T cells to tissue-specific self antigen, and may drive the autoimmune phenomenon in the salivary glands. We found that retinoblastoma-associated protein RbAp48 overexpression induces p53-mediated apoptosis in the salivary glands caused by estrogen deficiency. We demonstrated that transgenic (Tg) expression of RbAp48 resulted in the development of autoimmune exocrinopathy resembling Sjögren's syndrome (SS). CD4(+)T cell-mediated autoimmune lesions in the salivary glands were aggravated with age, in association with autoantibody productions. We obtained evidences that salivary epithelial cells can produce interferon-gamma (IFN-gamma) besides interleukin (IL)-18, which activates interferon regulatory factor-1 (IRF-1), and class II transactivator (CIITA). Indeed, the autoimmune lesions into Rag2(-/-) mice were induced by the adoptive transfer of lymph node cells from RbAp48-Tg mice. These results indicate a novel immunocompetent role of epithelial cells that can produce IFN-gamma, resulting in loss of local tolerance prior to developing gender-based autoimmunity. The studies reviewed the molecular mechanisms on the development of salivary gland autoimmunity, and gender-related differences in SS.
Kimi Yamakoshi, Akiko Takahashi, Fumiko Hirota, Rika Nakayama, Naozumi Ishimaru, Yoshiaki Kubo, J. David Mann, Masako Ohmura, Atsushi Hirao, Hideyuki Saya, Seiji Arase, Yoshio Hayashi, Kazuki Nakao, Mitsuru Matsumoto, Naoko Ohtani and Eiji Hara : Real-time in vivo imaging of p16Ink4a reveals cross-talk with p53, The Journal of Cell Biology, Vol.186, No.3, 393-407, 2009.
(要約)
Expression of the p16(Ink4a) tumor suppressor gene, a sensor of oncogenic stress, is up-regulated by a variety of potentially oncogenic stimuli in cultured primary cells. However, because p16(Ink4a) expression is also induced by tissue culture stress, physiological mechanisms regulating p16(Ink4a) expression remain unclear. To eliminate any potential problems arising from tissue culture-imposed stress, we used bioluminescence imaging for noninvasive and real-time analysis of p16(Ink4a) expression under various physiological conditions in living mice. In this study, we show that oncogenic insults such as ras activation provoke epigenetic derepression of p16(Ink4a) expression through reduction of DNMT1 (DNA methyl transferase 1) levels as a DNA damage response in vivo. This pathway is accelerated in the absence of p53, indicating that p53 normally holds the p16(Ink4a) response in check. These results unveil a backup tumor suppressor role for p16(Ink4a) in the event of p53 inactivation, expanding our understanding of how p16(Ink4a) expression is regulated in vivo.
Takeo Iwata, Masamichi Kuwajima, Akiko Sukeno, Naozumi Ishimaru, Yoshio Hayashi, Martin Wabitsch, Noriko Mizusawa, Mitsuo Itakura and Katsuhiko Yoshimoto : YKL-40 secreted from adipose tissue inhibits degradation of type I collagen., Biochemical and Biophysical Research Communications, Vol.388, No.3, 511-516, 2009.
(要約)
Obesity is considered a chronic low-grade inflammatory status and the stromal vascular fraction (SVF) cells of adipose tissue (AT) are considered a source of inflammation-related molecules. We identified YKL-40 as a major protein secreted from SVF cells in human visceral AT. YKL-40 expression levels in SVF cells from visceral AT were higher than in those from subcutaneous AT. Immunofluorescence staining revealed that YKL-40 was exclusively expressed in macrophages among SVF cells. YKL-40 purified from SVF cells inhibited the degradation of type I collagen, a major extracellular matrix of AT, by matrix metalloproteinase (MMP)-1 and increased rate of fibril formation of type I collagen. The expression of MMP-1 in preadipocytes and macrophages was enhanced by interaction between these cells. These results suggest that macrophage/preadipocyte interaction enhances degradation of type I collagen in AT, meanwhile, YKL-40 secreted from macrophages infiltrating into AT inhibits the type I collagen degradation.
Naozumi Ishimaru, Takagi Atuya, Kohashi Masayuki, Akiko Yamada, Rieko Arakaki, Kanno Jun and Yoshio Hayashi : Neonatal Exposure to Loe-Dose 2.3.7.8-Tetrachlorodibenzo-p-Dioxin Causes Autoimmunity Due to the Disruption of TCell Tolerance1, The Journal of Immunology, Vol.182, No.10, 6576-6586, 2009.
(要約)
Although 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been shown to influence immune responses, the effects of low-dose TCDD on the development of autoimmunity are unclear. In this study, using NFS/sld mice as a model for human Sjögren's syndrome, in which the lesions are induced by the thymectomy on day 3 after birth, the autoimmune lesions in the salivary glands, and in later phase, inflammatory cell infiltrations in the other organs were developed by neonatal exposure to nonapoptotic dosage of TCDD without thymectomy on day 3 after birth. We found disruption of thymic selection, but not thymic atrophy, in TCDD-administered mice. The endogenous expression of aryl hydrocarbon receptor in the neonatal thymus was significantly higher than that in the adult thymus, suggesting that the neonatal thymus may be much more sensitive to TCDD compared with the adult thymus. In addition, the production of T(H)1 cytokines such as IL-2 and IFN-gamma from splenic CD4(+) T cells and the autoantibodies relevant for Sjögren's syndrome in the sera from TCDD-exposed mice were significantly increased compared with those in control mice. These results suggest that TCDD/aryl hydrocarbon receptor signaling in the neonatal thymus plays an important role in the early thymic differentiation related to autoimmunity.
Rieko Arakaki, Ai Nagaoka, Naozumi Ishimaru, Akiko Yamada, Y Satoko and Yoshio Hayashi : Role of Plasmacytoid Dendric Cells for Aberrant Class Expression in Exocrine Glands from Estrogen-Deficient Mice of healthy Background, The American Journal of Pathology, Vol.174, No.5, 1715-1724, 2009.
(要約)
Although it has been well documented that aberrant major histocompatibility complex class II molecules may contribute to the development of autoimmune disorders, the precise mechanisms responsible for their tissue-specific expression remain unknown. Here we show that estrogen deficiency induces aberrant class II major histocompatibility complex expression in exocrine glands via interactions between epithelial cells and plasmacytoid dendritic cells. Relatively modest but functionally significant expression levels of major histocompatibility complex class II and class II transactivator molecules were observed in the exocrine glands of ovariectomized (Ovx) C57BL/6 (B6) mice, but were not seen in the exocrine glands of control B6 mice. We observed that the salivary dendritic cells adjacent to the apoptotic epithelial cells positive for terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling, were activated in Ovx mice, but were not activated in control mice. We obtained evidence that the salivary gland cells express both interferon regulatory factor-1 and class II transactivator type IV molecules in Ovx mice. Salivary gland cells from Ovx mice were also capable of inducing the activation of antigen-specific T cells from OT-II transgenic mice. These findings indicate that estrogen deficiency initiates class II transactivator type IV mRNA expression in exocrine glands via interactions between epithelial cells and plasmacytoid dendritic cells, suggesting that plasmacytoid dendritic cells play a pivotal role in gender-based autoimmune disorders in postmenopausal women.
Naozumi Ishimaru, Rieko Arakaki, Satoko Yoshida, Akiko Yamada, Sumihare Noji and Yoshio Hayashi : Expression of the retinoblastoma protein RbAp48 in exocrine glands leads to Sjögren's syndrome-like autoimmune exocrinopathy., The Journal of Experimental Medicine, Vol.205, No.12, 2915-2927, 2008.
(要約)
Although several autoimmune diseases are known to develop in postmenopausal women, the mechanisms by which estrogen deficiency influences autoimmunity remain unclear. Recently, we found that retinoblastoma-associated protein 48 (RbAp48) induces tissue-specific apoptosis in the exocrine glands depending on the level of estrogen deficiency. In this study, we report that transgenic (Tg) expression of RbAp48 resulted in the development of autoimmune exocrinopathy resembling Sjögren's syndrome. CD4(+) T cell-mediated autoimmune lesions were aggravated with age, in association with autoantibody productions. Surprisingly, we obtained evidence that salivary and lacrimal epithelial cells can produce interferon-gamma (IFN-gamma) in addition to interleukin-18, which activates IFN regulatory factor-1 and class II transactivator. Indeed, autoimmune lesions in Rag2(-/-) mice were induced by the adoptive transfer of lymph node T cells from RbAp48-Tg mice. These results indicate a novel immunocompetent role of epithelial cells that can produce IFN-gamma, resulting in loss of local tolerance before developing gender-based autoimmunity.
Yu Hikosaka, Takeshi Nitta, Izumi Ohigashi, Kouta Yano, Naozumi Ishimaru, Yoshio Hayashi, Mitsuru Matsumoto, Koichi Matsuo, Josef M Penninger, Hiroshi Takayanagi, Yoshifumi Yokota, Hisakata Yamada, Yasunobu Yoshikai, Jun-ichiro Inoue, Taishin Akiyama and Yousuke Takahama : The cytokine RANKL produced by positively selected thymocytes fosters medullary thymic epithelial cells that express autoimmune regulator., Immunity, Vol.29, No.3, 438-450, 2008.
(要約)
The thymic medulla provides a microenvironment where medullary thymic epithelial cells (mTECs) express autoimmune regulator and diverse tissue-restricted genes, contributing to launching self-tolerance. Positive selection is essential for thymic medulla formation via a previously unknown mechanism. Here we show that the cytokine RANK ligand (RANKL) was produced by positively selected thymocytes and regulated the cellularity of mTEC by interacting with RANK and osteoprotegerin. Forced expression of RANKL restored thymic medulla in mice lacking positive selection, whereas RANKL perturbation impaired medulla formation. These results indicate that RANKL produced by positively selected thymocytes is responsible for fostering thymic medulla formation, thereby establishing central tolerance.
We reported three cases of Mikuhcz's disease. Mikulicz's disease was diagnosed by the following criteria: 1) symmetrical enlargement of the lacrimal and salivary glands lasting at least 3 months', 2) the lacrimal and salivary glands were microscopically involved in the infiltration of inflammatory cells, 3) sarcoidosis and other lymphoproliferative diseases were excluded. Three patients with Mikulicz's disease were all characterized by elevated IgG4 concentration in the serum and prominent infiltration of IgG4-positive plasmacytes in the minor salivary glands. On the contrary, there were no findings indicating sicca syndrome or anti-SS-A and SS-B antibodies, which characterize Sjögren syndrome. It is suggested that Mikulicz's disease is an IgG4-related plasmacytec disease, with a pathogenesis differing from that of Sjögren syndrome.
Nao Kinouchi, Yutaka Ohsawa, Naozumi Ishimaru, Hideyo Ohuchi, Y Sunada, Yoshio Hayashi, Yukiho Tanimoto, Keiji Moriyama and Sumihare Noji : Atelocollagen-mediated local and systemic applications of myostatin-targeting siRNA increase skeletal muscle mass., Gene Therapy, Vol.15, No.15, 1126-1130, 2008.
(要約)
RNA interference (RNAi) offers a novel therapeutic strategy based on the highly specific and efficient silencing of a target gene. Since it relies on small interfering RNAs (siRNAs), a major issue is the delivery of therapeutically active siRNAs into the target tissue/target cells in vivo. For safety reasons, strategies based on vector delivery may be of only limited clinical use. The more desirable approach is to directly apply active siRNAs in vivo. Here, we report the effectiveness of in vivo siRNA delivery into skeletal muscles of normal or diseased mice through nanoparticle formation of chemically unmodified siRNAs with atelocollagen (ATCOL). ATCOL-mediated local application of siRNA targeting myostatin, a negative regulator of skeletal muscle growth, in mouse skeletal muscles or intravenously, caused a marked increase in the muscle mass within a few weeks after application. These results imply that ATCOL-mediated application of siRNAs is a powerful tool for future therapeutic use for diseases including muscular atrophy.
Naozumi Ishimaru, Akiko Yamada, Masayuki Kohashi, Rieko Arakaki, Tetsuyuki Takahashi, Keisuke Izumi and Yoshio Hayashi : Development of Inflammatory Bowel Disease in Long-Evans Cinnamon Rats Based on CD4+CD25+Foxp3+ Regulatory T Cell Dysfunction, The Journal of Immunology, Vol.180, No.10, 6997-7008, 2008.
(要約)
A mutant strain with defective thymic selection of the Long-Evans Cinnamon (LEC) rat was found to spontaneously develop inflammatory bowel disease (IBD)-like colitis. The secretion of Th1-type cytokines including IFN-gamma and IL-2 from T cells of mesenteric lymph node cells (MLNs) and lamina propria mononuclear cells, but not spleen cells, in LEC rats was significantly increased more than that of the control Long-Evans Agouti rats through up-regulated expression of T-bet and phosphorylation of STAT-1 leading to NF-kappaB activation. In addition, the number of CD4(+)CD25(+)Foxp3(+) regulatory T (Treg) cells of the thymus, MLNs, and lamina propria mononuclear cells from LEC rats was significantly reduced, comparing with that of the control rats. Moreover, bone marrow cell transfer from LEC rats into irradiated control rats revealed significantly reduced CD25(+)Foxp3(+) Treg cells in thymus, spleen, and MLNs compared with those from control rats. Indeed, adoptive transfer with T cells of MLNs, not spleen cells, from LEC rats into SCID mice resulted in the development of inflammatory lesions resembling the IBD-like lesions observed in LEC rats. These results indicate that the dysfunction of the regulatory system controlled by Treg cells may play a crucial role in the development of IBD-like lesions through up-regulated T-bet, STAT-1, and NF-kappaB activation of peripheral T cells in LEC rats.
Masayuki Kohashi, Naozumi Ishimaru, Rieko Arakaki and Yoshio Hayashi : Effective Treatment With Oral Administration of Rebamipide in a Model of Sjogren's Syndrome, Arthritis and Rheumatism, Vol.58, No.2, 389-400, 2008.
(要約)
To determine whether oral administration of rebamipide, a mucosal protective agent, is effective in the treatment of Sjögren's syndrome (SS) in the NFS/sld mouse model of the disease. NFS/sld mice were given daily oral doses of rebamipide (0.3 mg/kg of body weight or 3 mg/kg) or vehicle alone starting from the age of 4 weeks to the age of 8 weeks. The volume of saliva and tears was monitored during and after treatment. After the final dose, histologic features of the tissues, TUNEL+ apoptotic duct cells in affected glands, T cell and cytokine function, and levels of immunoglobulin isotypes and serum autoantibodies were examined. The 3-mg/kg dose of rebamipide prevented the development of autoimmune lesions. The average volume of saliva in rebamipide-treated mice was significantly higher than that in control mice. We found decreased TUNEL+ apoptotic duct cells in the salivary and lacrimal glands of rebamipide-treated mice as compared with control mice. Rebamipide treatment suppressed the activation of CD4+ T cells and Th1 cytokines (interleukin-2, interferon-gamma) associated with impaired NF-kappaB activity. Production of serum autoantibodies, IgM, and IgG1 was clearly inhibited. Our findings demonstrate the efficacy of oral administration of rebamipide in the treatment of SS. Rebamipide represents a new therapeutic strategy for the treatment of patients with sicca symptoms caused by SS, as well as for patients with other diseases.
Kazuhito Satomura, Reiko Tokuyama, Tetsuya Yuasa, Y Yamasaki, Seiko Tatehara, Naozumi Ishimaru, Yoshio Hayashi and Masaru Nagayama : Possible involvement of stem cell factor and endothelin-1 in the emergence of pigmented squamous cell carcinoma in oral mucosa., Journal of Oral Pathology & Medicine, Vol.36, No.10, 621-624, 2007.
(要約)
We present here the clinical, morphological and immunohistochemical features of a pigmented squamous cell carcinoma (SCC) in the oral mucosa of the hard palate of a 76-year-old Japanese man. He underwent a partial resection of the maxilla subsequent to radiotherapy. The tumor was typical, moderately well-differentiated SCC but had many melanocytes (melanocytosis) within it. Immunohistochemical analysis for stem cell factor (SCF) and endothelin-1, both of which are known to stimulate proliferation and differentiation of melanocytes, revealed prominent expression of both factors in the neoplastic squamous cells of the pigmented SCC, while the non-pigmented oral SCC showed little sign of either factor. These findings strongly suggest that SCF and endothelin-1 secreted by neoplasmic squamous cells are involved in the emergence of a rare variant of oral SCC.
Naoko Ohtani, Yuko Imamura, Kimi Yamakoshi, Fumiko Hirota, Rika Nakayama, Yoshiaki Kubo, Naozumi Ishimaru, Akiko Takahashi, Atsushi Hirao, Takatsune Shimizu, David J. Mann, Hideyuki Saya, Yoshio Hayashi, Seiji Arase, Mitsuru Matsumoto, Nakao Kazuki and Eiji Hara : Visualizing the dynamics of p21 (Wafl/Cip1)cyclin-dependent kinase inhibitor expression in living animals., Proceedings of the National Academy of Sciences of the United States of America, Vol.104, No.38, 15034-15039, 2007.
(要約)
Although the role of p21(Waf1/Cip1) gene expression is well documented in various cell culture studies, its in vivo roles are poorly understood. To gain further insight into the role of p21(Waf1/Cip1) gene expression in vivo, we attempted to visualize the dynamics of p21(Waf1/Cip1) gene expression in living animals. In this study, we established a transgenic mice line (p21-p-luc) expressing the firefly luciferase under the control of the p21(Waf1/Cip1) gene promoter. In conjunction with a noninvasive bioluminescent imaging technique, p21-p-luc mice enabled us to monitor the endogenous p21(Waf1/Cip1) gene expression in vivo. By monitoring and quantifying the p21(Waf1/Cip1) gene expression repeatedly in the same mouse throughout its entire lifespan, we were able to unveil the dynamics of p21(Waf1/Cip1) gene expression in the aging process. We also applied this system to chemically induced skin carcinogenesis and found that the levels of p21(Waf1/Cip1) gene expression rise dramatically in benign skin papillomas, suggesting that p21(Waf1/Cip1) plays a preventative role(s) in skin tumor formation. Surprisingly, moreover, we found that the level of p21(Waf1/Cip1) expression strikingly increased in the hair bulb and oscillated with a 3-week period correlating with hair follicle cycle progression. Notably, this was accompanied by the expression of p63 but not p53. This approach, together with the analysis of p21(Waf1/Cip1) knockout mice, has uncovered a novel role for the p21(Waf1/Cip1) gene in hair development. These data illustrate the unique utility of bioluminescence imaging in advancing our understanding of the timing and, hence, likely roles of specific gene expression in higher eukaryotes.
Takashi Izawa, Naozumi Ishimaru, Keiji Moriyama, Masayuki Kohashi, Rieko Arakaki and Yoshio Hayashi : Crosstalk between RANKL and Fas signaling in dendritic cells controls immune tolerance, Blood, Vol.110, No.1, 242-250, 2007.
(要約)
Although receptor activator of nuclear factor (NF)-kappaB ligand (RANKL) signaling has been shown to prolong the survival of mature dendritic cells (DCs), the association of RANKL pathway with Fas-mediated apoptosis is obscure. Here, we found that bone marrow-derived DCs (BMDCs) from the Fas-deficient strain MRL/lpr mice, could survive much longer than normal DCs. The expressions of Bcl-x and Bcl-2 and the nuclear transport of NF-kappaB of RANKL-stimulated BMDCs from MRL/lpr mice were significantly up-regulated. By contrast, Fas expression of BMDCs from normal C57BL/6 and MRL(+/+) mice was increased by RANKL stimulation, and an enhanced DC apoptosis was found when stimulated with both RANKL and anti-Fas mAb, which was associated with activation of caspase-3 and caspase-9. Furthermore, the expression of FLIP(L), an inhibitory molecule against Fas-mediated apoptosis, in normal DCs was significantly decreased by RANKL and anti-Fas mAb. Indeed, the adoptive transfer of RANKL-stimulated DCs resulted in rapid acceleration of autoimmunity in MRL/lpr recipients. These findings indicate that the crosstalk between RANKL and Fas signaling in DCs might control immune tolerance.
Naozumi Ishimaru, Hidehiro Kishimoto, Yoshio Hayashi and Jonathan Sprent : Regulation of naive T cell function by the NF-κB2 pathway, Nature Immunology, Vol.7, No.7, 763-772, 2006.
(要約)
T cell activation involves the orchestration of several signaling pathways, including that of the 'classical' transcription factor NF-kappaB components NF-kappaB1-RelA. The function of the 'nonclassical' NF-kappaB2-RelB pathway is less clear, although T cells lacking components of this pathway have activation defects. Here we show that mice deficient in NF-kappaB-inducing kinase have a complex phenotype consisting of immunosuppression mediated by CD25(-)Foxp3(-) memory CD4(+) cells and, in the absence of those cells, hyper-responsive naive CD4(+) T cells, which caused autoimmune lesions after adoptive transfer into hosts deficient in recombination-activating genes. Biochemical studies indicated involvement of a cell-intrinsic mechanism in which NF-kappaB2 (p100) limits nuclear translocation of NF-kappaB1-RelA and thereby functions as a regulatory 'brake' for the activation of naive T cells.
Nobuhiko Katunuma, Quang Trong Le, Etsuko Murata, Atsushi Matsui, Eiji Majima, Naozumi Ishimaru, Yoshio Hayashi and Atsushi Ohashi : A novel apoptosis cascade mediated by lysosomal lactoferrin and its participation in hepatocyte apoptosis induced by D-galactosamine., FEBS Letters, Vol.580, No.15, 3699-3705, 2006.
(要約)
A new apoptosis cascade mediated by lysosomal lactoferrin was found in apoptotic liver induced by d-galactosamine. Caspase-3 and lactoferrin were increased in the apoptotic liver cytoplasm and serum transaminases were elevated. Recombinant lactoferrin stimulated procaspase-3 processing at 10(-6)-10(-7)M to an extent similar to that by granzyme B in vitro. Lactoferrin changed procaspase-3 structure susceptible to the processing. Synthetic peptide Y(679)-K(695) in lactoferrin molecule inhibited the processing of procaspase-3 by lactoferrin. Lactoferrin in lysosomes was decreased and lactoferrin released into cytoplasm was increased quantitatively in d-galactosamine induced apoptotic liver, and procaspase-3 in cytoplasm was processed to caspase-3.
Masayuki Azuma, Yuki Ashida, Tetsuya Tamatani, Katsumi Motegi, Natsumi Takamaru, Naozumi Ishimaru, Yoshio Hayashi and Mitsunobu Sato : Cepharanthin,a biscoclaurine alkaloid,prevents destruction of acinar tissues in murine Sjogren's syndrome., The Journal of Rheumatology, Vol.33, No.5, 912-920, 2006.
(要約)
Our previous study suggested that suppression by cepharanthin of tumor necrosis factor-a (TNF-a)-induced matrix metalloproteinase-9 (MMP-9) could prevent destruction of the acinar structure in the salivary glands of patients with Sjögren's syndrome (SS). In this study, we observed that in vivo administration of cepharanthin prevented severe damage to acinar tissues in the murine model of human SS. Cepharanthin was intraperitoneally administered to thymectomized female NFS/sld mice. Inflammatory lesions in the salivary and lacrimal glands were then examined histologically. Expression of phosphorylated IkB-a, MMP-9, and type IV collagen was analyzed immunohistochemically. The apoptotic cell death of acinar cells was determined. Although extensive mononuclear cell infiltration and destruction of acinar tissue in salivary and lacrimal glands were observed in control mice, significant improvement of these lesions was evident in mice treated with cepharanthin. Immunohistochemical analysis revealed that p65, phosphorylated IkB-a, and MMP-9 were more strongly stained in the acinar cells of control mice than in cepharanthin-treated mice. Although no staining for type IV collagen was observed in the acinar tissues of control mice, continuity of staining for type IV collagen was observed in acinar tissues of cepharanthin-treated mice. Destruction of acinar tissues was attributed to the induction of apoptosis, suggesting that cepharanthin inhibits apoptosis by suppressing phosphorylation of IkB-a, followed by prevention of MMP-9 activation. Our findings suggest that cepharanthin may be a promising agent for use in preventing destruction of acinar tissues in murine SS.
Naoto Suzue, Takeshi Nikawa, Yuko Onishi, Chiharu Yamada, Katsuya Hirasaka, Takayuki Ogawa, Harumi Furochi, Hirofumi Kosaka, Kazumi Ishidoh, Gu Hua, Shin'ichi Takeda, Naozumi Ishimaru, Yoshio Hayashi, Hironori Yamamoto, Kyoichi Kishi and Natsuo Yasui : Ubiqitin Ligase Cbl-b Downregulates Bone Formation Through Suppression of IGF-I Signaling in Osteoblasts During Denervation, Journal of Bone and Mineral Research, Vol.21, No.5, 722-734, 2006.
(要約)
Unloading can prevent bone formation by osteoblasts. To study this mechanism, we focused on a ubiquitin ligase, Cbl-b, which was highly expressed in osteoblastic cells during denervation. Our results suggest that Cbl-b may mediate denervation-induced osteopenia by inhibiting IGF-I signaling in osteoblasts. Unloading, such as denervation (sciatic neurectomy) and spaceflight, suppresses bone formation by osteoblasts, leading to osteopenia. The resistance of osteoblasts to growth factors contributes to such unloading-mediated osteopenia. However, a detailed mechanism of this resistance is unknown. We first found that a RING-type ubiquitin ligase, Cbl-b, was highly expressed in osteoblastic cells after sciatic neurectomy in mice. In this study, we reasoned that Cbl-b played an important role in the resistance of osteoblasts to IGF-I. Cbl-b-deficient (Cbl-b(-/-)) or wildtype (Cbl-b(+/+)) mice were subjected to sciatic neurectomy. Bone formation in these mice was assessed by calcein labeling and histomorphometric analyses. We examined IGF-I signaling molecules in femora of these mice by Western blot and immunohistochemical analyses. We also examined the mitogenic response of Cbl-b-overexpressing or -deficient osteoblastic cells to various growth factors. In Cbl-b(+/+) mice, denervation decreased femur mass and bone formation, whereas it increased the expression of Cbl-b protein in osteoprogenitor cells and in osteocalcin-positive cells (osteoblastic cells) in hindlimb bone. In contrast, in Cbl-b(-/-) mice, bone mass and bone formation were sustained during denervation. Denervation inhibited the mitogenic response of osteoprogenitor cells most significantly to IGF-I. Therefore, we focused on Cbl-b-mediated modification of IGF-I signaling. Denervation decreased the amounts of insulin receptor substrate-1 (IRS-1), phosphatidly inositol 3-phosphate kinase (PI3K), and Akt-1 proteins in femora of Cbl-b(+/+) mice, whereas the amounts of these IGF-I signaling molecules in femora of Cbl-b(-/-) mice were constant after denervation. On a cellular level, primary osteoblastic cells from Cbl-b(-/-) mice were more stimulated to proliferate by IGF-I treatment compared with those from Cbl-b(+/+) mice. Furthermore, overexpression of Cbl-b increased ubiquitination and degradation of IRS-1 in primary Cbl-b(-/-) osteoblastic cells, leading to their impaired mitogenic response to IGF-I. These results suggest that Cbl-b induces resistance of osteoblasts to IGF-I during denervation by increasing IRS-1 degradation and that Cbl-b-mediated modification of IGF-I signaling may contribute to decreased bone formation during denervation.
(キーワード)
Animals / Base Sequence / Blotting, Western / Bone Development / Cells, Cultured / DNA Primers / Denervation / Down-Regulation / Hydrolysis / 免疫組織化学 (immunohistochemistry) / Insulin-Like Growth Factor I / Mice / Mice, Inbred C57BL / Osteoblasts / Protein Binding / Reverse Transcriptase Polymerase Chain Reaction / シグナル伝達 (signal transduction) / Ubiquitin / Ubiquitin-Protein Ligases
Naozumi Ishimaru, Rieko Arakaki, Fumie Omotehara, Kouichi Yamada, Kenji Mishima, Ichiro Saito and Yoshio Hayashi : Novel role for RbAp48 in tissue-specific,estrogen deficiency-dependent apoptosis in the exocrine glands., Molecular and Cellular Biology, Vol.26, No.8, 2924-2935, 2006.
(要約)
Although tissue-specific apoptosis in the exocrine glands in estrogen-deficient mice may contribute to the development of autoimmune exocrinopathy, the molecular mechanism responsible for tissue-specific apoptosis remains obscure. Here we show that RbAp48 overexpression induces p53-mediated apoptosis in the exocrine glands caused by estrogen deficiency. RbAp48-inducible transfectant results in rapid apoptosis with p53 phosphorylation (Ser9) and alpha-fodrin cleavage. Reducing the expression of RbAp48 through small interfering RNA inhibits the apoptosis. Prominent RbAp48 expression with apoptosis was observed in the exocrine glands of C57BL/6 ovariectomized (OVX) mice but not in OVX estrogen receptor alpha(-/-), p53(-/-), and E2F-1(-/-) mice. Indeed, transgenic expression of the RbAp48 gene induced apoptosis in the exocrine glands but not in other organs. These findings indicate that estrogen deficiency initiates p53-mediated apoptosis in the exocrine gland cells through RbAp48 overexpression and exerts a possible gender-based risk of autoimmune exocrinopathy in postmenopausal women.
Nobuhiko Katumuma, Atsushi Ohashi, Etsuko Sano, Naozumi Ishimaru, Yoshio Hayashi and Etsuko Murata : Catechin derivatives:specific inhibitor for caspases-3,7and 2,and the prevention of apoptosis at the cell and animal levels., FEBS Letters, Vol.580, No.3, 741-746, 2006.
(要約)
Tea-catechin derivatives are shown to inhibit activities of caspases-3, 2 and 7 in vitro, and prevented experimental apoptosis at the cell and animal levels. Epigallo-catechin-gallate showed the strongest inhibition at 1 x 10(-7)M to these caspases, but cysteine cathepsins and caspase-8 were not inhibited. Caspase-3 inhibition showed a 2nd-order allosteric-type, but the inhibition of caspases-2 and 7 showed a non-competitive-type. The apoptosis-test using cultured HeLa cells was inhibited by these catechins. In rat hepatocytes, apoptosis was induced by d-galactosamine in vivo. In this case, caspase-3 activity in the cytoplasm, the serum aminotransferases and dUTP nick formation detected by TUNNEL-staining were effects, and these elevations were suppressed by administration of catechin.
Hirotsugu Kurobe, Cunian Liu, Tomoo Ueno, Fumi Saito, Izumni Ohigashi, Natalle Seach, Rieko Arakaki, Yoshio Hayashi, Tetsuya Kitagawa, Martin Lipp, Richard L. Boyd and Yousuke Takahama : CCR7-Dependent Cortex-to-Medulla Migration of Positively Selected Thymocytes Is Essential for Establishing Central Tolerance, Immunity, Vol.24, No.2, 165-177, 2006.
(要約)
Immature CD4+CD8+ thymocytes, which are generated in the thymic cortex, are induced upon positive selection to differentiate into mature T lymphocytes and relocate to the thymic medulla. It was recently shown that a chemokine signal via CCR7 is essential for the cortex-to-medulla migration of positively selected thymocytes in the thymus. However, the role of the cortex-to-medulla migration in T cell development and selection has remained unclear. The present study shows that the developmental kinetics and the thymic export of mature thymocytes were undisturbed in adult mice lacking CCR7 or its ligands (CCR7L). The inhibition of sphingosine-1-phosphate-mediated lymphocyte egress from the thymus led to the accumulation of mature thymocytes in the cortex of CCR7- or CCR7L-deficient mice, unlike the accumulation in the medulla of normal mice, thereby suggesting that mature thymocytes may be exported directly from the cortex in the absence of CCR7 signals. However, the thymocytes that were generated in the absence of CCR7 or CCR7L were potent in causing autoimmune dacryoadenitis and sialadenitis in mice and were thus incapable of establishing central tolerance to organ-specific antigens. These results indicate that CCR7-mediated cortex-to-medulla migration of thymocytes is essential for establishing central tolerance rather than for supporting the maturation or export of thymocytes.
Kumi Obara, Fumio Ide, Kenji Mishima, Hiroko Inoue, Hiroyuki Yamada, Yoshio Hayashi and Ichiro Saito : Biological and oncogenic properties of p53-deficient salivary gland epithelial cells with particular emphasis on stromal-epithelial interactions in tumorigenesis., Pathobiology, Vol.73, No.5, 261-270, 2006.
(要約)
To understand the salivary gland pathobiology, we established an immortalized duct/basal cell line (MSE) from the submandibular glands of p53-deficient mice. A variety of culture assays and xenograft experiments were conducted. Cellular characteristics were analyzed using histological, immunohistochemical, ultrastructural, and molecular techniques. Inoculation of a mixture of MSE and Matrigel reconstructed polarized ducts whereas cotransplantation of MSE with both Matrigel and NIH3T3 (3T3) cells developed mixed tumors of adenoma and sarcoma. A daughter adenoma line (MSA) showed some transformed phenotype in vitro, but was marginally tumorigenic in vivo. Notably, pleomorphic adenoma gene 1 (PLAG1) was expressed in MSA but not in MSE. As compared with MSE, MSA showed higher levels of insulin-like growth factor-I receptor (IGF-IR). Interestingly, 3T3 sarcoma secreted insulin-like growth factor-II (IGF-II), while MSA did not. The intrinsic tumorigenic programs of p53 null salivary epithelium are promoted by 3T3 sarcoma-derived IGF-IIin a paracrine manner through overexpression of PLAG1 and IGF-IR.
Katsushi Miyazaki, Noriaki Takeda, Naozumi Ishimaru, Fumie Omotehara, Rieko Arakaki and Yoshio Hayashi : Analysis of in vivo role of alpha-fodrin autoantigen in primary Sjogren's syndrome, The American Journal of Pathology, Vol.167, No.4, 1051-1059, 2005.
(要約)
The alpha-fodrin N-terminal portion (AFN) autoantigen mediates in vivo immunoregulation of autoimmune responses in primary Sjögren's syndrome (SS). We further examined this process and found that cleavage products of AFN were frequently detected in the salivary gland duct cells of SS patients. In in vitro studies using human salivary gland HSY cells, anti-Fas-induced apoptosis resulted in specific cleavage of alpha-fodrin into the 120-kd fragment, in association of alpha-fodrin with mu-calpain, and activation of caspase 3. Significant proliferative responses against AlphaFN autoantigen were observed in the peripheral blood mononuclear cells (PBMCs) from SS patients with higher pathological score (grade 4) and with short duration from onset (within 5 years). In vivo roles of AFN peptides were investigated using PBMCs from patients with SS, systemic lupus erythematosus, and rheumatoid arthritis. Significant proliferative T-cell responses of PBMCs to AFN peptide were detected in SS but not in systemic lupus erythematosus or rheumatoid arthritis. AFN peptide induced Th1-immune responses and accelerated down-regulation of Fas-mediated T-cell apoptosis in SS. Our data further elucidate the in vivo role of AFN autoantigen on the development of SS and suggest that the AFN autoantigen is a novel participant in peripheral tolerance.
Noriyuki Kuroda, Tasuku Mitani, Naoki Takeda, Naozumi Ishimaru, Rieko Arakaki, Yoshio Hayashi, Yoshimi Bando, Keisuke Izumi, Takeshi Takahashi, Takashi Nomura, Shimon Sakaguchi, Tomoo Ueno, Yousuke Takahama, Daisuke Uchida, Shijie Sun, Fumiko Kajiura, Yasuhiro Mouri, Hongwei Han, Akemi Matsushima, Gen Yamada and Mitsuru Matsumoto : Development of autoimmunity against transcriptionally unrepressed target antigen in the thymus of Aire-deficient mice, The Journal of Immunology, Vol.174, No.4, 1862-1870, 2005.
(要約)
Autoimmune regulator (AIRE) gene mutation is responsible for the development of organ-specific autoimmune disease with monogenic autosomal recessive inheritance. Although Aire has been considered to regulate the elimination of autoreactive T cells through transcriptional control of tissue-specific Ags in thymic epithelial cells, other mechanisms of AIRE-dependent tolerance remain to be investigated. We have established Aire-deficient mice and examined the mechanisms underlying the breakdown of self-tolerance. The production and/or function of immunoregulatory T cells were retained in the Aire-deficient mice. The mice developed Sjogren's syndrome-like pathologic changes in the exocrine organs, and this was associated with autoimmunity against a ubiquitous protein, alpha-fodrin. Remarkably, transcriptional expression of alpha-fodrin was retained in the Aire-deficient thymus. These results suggest that Aire regulates the survival of autoreactive T cells beyond transcriptional control of self-protein expression in the thymus, at least against this ubiquitous protein. Rather, Aire may regulate the processing and/or presentation of self-proteins so that the maturing T cells can recognize the self-Ags in a form capable of efficiently triggering autoreactive T cells. With the use of inbred Aire-deficient mouse strains, we also demonstrate the presence of some additional factor(s) that determine the target-organ specificity of the autoimmune disease caused by Aire deficiency.
Miriam Entesarian, Hans Matsson, Joakim Klar, Birgitta Bergendal, Lena Olson, Rieko Arakaki, Yoshio Hayashi, Hideyo Ohuchi, Babak Falahat, Isinev Anne Bolstad, Roland Jonsson, Marie Wahren-Herlenius and Niklas Dahl : Mutations in the gene encoding fibroblast growth factor 10 are associated with aplasia of lacrimal and salivary glands, Nature Genetics, Vol.37, No.2, 125-127, 2005.
(要約)
Autosomal dominant aplasia of lacrimal and salivary glands (ALSG; OMIM 180920 and OMIM 103420) is a rare condition characterized by irritable eyes and dryness of the mouth. We mapped ALSG to 5p13.2-5q13.1, which coincides with the gene fibroblast growth factor 10 (FGF10). In two extended pedigrees, we identified heterozygous mutations in FGF10 in all individuals with ALSG. Fgf10(+/-) mice have a phenotype similar to ALSG, providing a model for this disorder. We suggest that haploinsufficiency for FGF10 during a crucial stage of development results in ALSG.
Masaru Kobayashi, Natsuo Yasui, Naozumi Ishimaru, Rieko Arakaki and Yoshio Hayashi : Development of Autoimmune Arthritis With Aging Via Bystander T Cell Activation in the Mouse Model of Sjogren's Syndrome, Arthritis and Rheumatism, Vol.50, No.12, 3974-3984, 2004.
(要約)
A wide spectrum of extraglandular manifestations may occur in patients with Sjogren's syndrome (SS), but the mechanisms responsible for in vivo progression are still obscure. We undertook this study to evaluate the age-related changes during the development of extraglandular autoimmune lesions, including arthritis, in the murine model of primary SS, and to evaluate the possible relationship between age-related disturbance of activation-induced cell death and the in vivo kinetics against autoantigens. A total of 126 NFS/sld mice were investigated at ages 2, 4, 6, 10, 12, 18, 20, and 24 months. Cytokine production was tested using culture supernatants from anti-CD3 monoclonal antibody-stimulated T cells. Anti-single-stranded DNA (anti-ssDNA) antibodies, Ig isotypes (IgG1, IgG2a), rheumatoid factor (RF), and anti-type II collagen (anti-CII) antibodies were detected by enzyme-linked immunosorbent assay. Proliferative T cell responses against each of 3 recombinant alpha-fodrin proteins and against CII were analyzed. Autoimmune arthritis developed in SS model mice until age 24 months. Significant elevations in serum levels of RF, anti-ssDNA antibodies, and anti-CII antibodies were found in aging SS model mice. A high titer of serum autoantibodies against alpha-fodrin fragments (containing different epitopes that were originally identified in primary SS model mice) was frequently detected in young and aged SS model mice. Moreover, we found that alpha-fodrin autoantigen induced Th1 immune responses and accelerated disturbance of Fas-mediated T cell apoptosis in aged SS model mice. These results indicate that age-related disturbance of activation-induced cell death via bystander T cell activation may play a crucial role in the development of autoimmune arthritis in a murine model of SS.
Yuki Hayashi, Naozumi Ishimaru, Rieko Arakaki, Shin-ichi Tsukumo, Hitomi Fukui, Kenji Kishihara, Hiroshi Shiota, Koji Yasutomo and Yoshio Hayashi : Effective Treatment of a Mouse Model of Sjogren's Syndrome With Eyedrop Administrasiton of Anti-CD4 Monoclonal Antibody, Arthritis and Rheumatism, Vol.50, No.9, 2903-2910, 2004.
(要約)
To determine whether eyedrop administration of an anti-CD4 monoclonal antibody (mAb) is effective in the treatment of Sjögren's syndrome (SS) using a mouse model of the disease. The anti-CD4 mAb was administered daily into the eyes of mice with SS from ages 4 to 8 weeks or ages 10 to 12 weeks. During treatment, tear volume was monitored and after final treatment, histologic features of the lacrimal and salivary glands, the phenotypes and function of T cells, and serum titers of anti-alpha-fodrin antibody were examined. Eyedrop administration of anti-CD4 mAb before the onset of SS prevented the autoimmune pathology seen in the lacrimal glands but not that in the salivary glands. Furthermore, eyedrop administration of anti-CD4 mAb after the development of SS inhibited mononuclear cell infiltration and the destruction of parenchyma only in the lacrimal glands. Eyedrop administration of anti-CD4 mAb suppressed the local activation of CD4+ T cells rather than deleting CD4+ T cells, which reduced the expansion of pathologic CD4+ T cells against alpha-fodrin. These results demonstrate the remarkable efficacy of anti-CD4 mAb eyedrops in the treatment of SS eye symptoms, which illustrates a new antibody-based therapeutic strategy for patients with eye problems caused by SS as well as other diseases.
Nobuhiko Katumuma, Etsuko Murata, T Quang Le, Yoshio Hayashi and Atsushi Ohashi : Newapoptosis cascade mediated by lysosomal enzyme and its protection by epigallo-catechin gallate, Advances in Enzyme Regulation, Vol.44, No.1, 1-10, 2004.
(要約)
We found a novel procaspase-3 activating cascade mediated by lysosomal enzyme. The activating enzyme of procaspase-3, named lysoapoptase having the molecular weight of 78kDa was determined to be a lactoferrin located in the lysosome. Recombinant lactoferrin accelerated the processing of procaspase-3 to form active caspase-3 in vitro. D-Galactosamine is a well-known inducer of hepatocyte apoptosis. The caspase-3 which plays a common central role in the final step of various apoptosis cascades, was dramatically increased in the cytoplasm by the d-galactosamine administration in vivo. When D-galactosamine was administrated as a death signal in vivo, the lysosomal lactoferrin was released into the cytoplasm and procaspase-3 located in the cytoplasm was processed to form active caspase-3. The cotreatment of epigallo-catechin gallate resulted in the complete protection of the hepatocyte apoptosis suppressing the increases of caspase-3 in the cytoplasm. The caspase-3 activity was also inhibited directly by the epigallo-catechin gallate. Therefore, all apoptosis cascades mediated by caspase-3 should be suppressed by epigallo-catechin gallate. The caspase-3 activity was not only directly inhibited by epigallo-catechin gallate in vitro, but the release of lactoferrin from the lysosomes into the cytoplasm was also suppressed by epigallo-catechin gallate treatment in vivo. Therefore, the apoptosis induction was suppressed at the two successive steps by cotreatment of epigallo-catechin gallate in vivo.
Toshiaki Maruyama, Ichiro Saito, Yoshio Hayashi, Elizabeth Kompfner, Robert I. Fox, Dennis R. Burton and Henrik J. Ditzel : Molecular Analysis of the Human Autoantibody Response to α-Fodrin in Sjogren Syndrome Reveals Novel Apoptosis-Induced Specificity, The American Journal of Pathology, Vol.165, No.1, 53-61, 2004.
(要約)
Lymphocyte infiltration of salivary and lacrimal glands leading to diminished secretion and gland destruction as a result of apoptosis is thought to be pivotal in the pathogenesis of Sjögren's syndrome (SS). The cytoskeletal protein alpha-fodrin is cleaved during this apoptotic process, and a strong antibody (Ab) response is elicited to a 120-kd fragment of cleaved alpha-fodrin in the majority of SS patients, but generally not in other diseases in which apoptosis also occurs. Little is known about the anti-alpha-fodrin autoantibody response on a molecular level. To address this issue, IgG phage display libraries were generated from the bone marrow of two SS donors and a panel of anti-alpha-fodrin IgGs was isolated by selection on alpha-fodrin immunoblots. All of the human monoclonal Abs (hmAbs) reacted with a 150-kd fragment and not with the 120-kd fragment or intact alpha-fodrin, indicating that the epitope recognized became exposed after alpha-fodrin cleavage. Analysis of a large panel of SS patients (defined by the strict San Diego diagnostic criteria) showed that 25% of SS sera exhibited this 150-kd alpha-fodrin specificity. The hmAbs stained human cultured salivary acinar cells and the staining was redistributed to surface blebs during apoptosis. They also stained inflamed acinar/ductal epithelial cells in SS salivary tissue biopsies, and only partially co-localized with monoclonal Abs recognizing the full-length alpha-fodrin. Our study shows that in SS patients, neoepitopes on the 150-kd cleaved product of alpha-fodrin become exposed to the immune system, frequently eliciting anti-150-kd alpha-fodrin Abs in addition to the previously reported anti-120-kd Abs. The anti-150-kd alpha-fodrin hmAbs may serve as valuable reagents for the study of SS pathogenesis and diagnostic analyses of SS salivary gland tissue.
The aims of this study were to evaluate the in vivo effects of estrogen deficiency in MRL/lpr mice as a model for rheumatoid arthritis and to analyze the possible relationship between immune dysregulation and receptor activator of nuclear factor-kappaB ligand (RANKL)-mediated osteoclastogenesis. Experimental studies were performed in ovariectomized (Ovx)-MRL/lpr, Ovx-MRL+/+, sham-operated-MRL/lpr, and sham-operated-MRL+/+ mice. Severe autoimmune arthritis developed in younger Ovx-MRL/lpr mice until 24 wk of age, whereas these lesions were entirely recovered by pharmacological levels of estrogen administration. A significant elevation in serum rheumatoid factor, anti-double-stranded DNA, and anti-type II collagen was found in Ovx-MRL/lpr mice and recovered in mice that underwent estrogen administration. A high proportion of CD4(+) T cells bearing RANKL was found, and an enhanced expression of RANKL mRNA and an impaired osteoprotegerin mRNA was detected in the synovium. An increase in both osteoclast formation and bone resorption pits was found. These results indicate that estrogen deficiency may play a crucial role in acceleration of autoimmune arthritis associated with RANKL-mediated osteoclastogenesis in a murine model for rheumatoid arthritis.
Hideko Endo, Mitsuyoshi Hirokawa, Naozumi Ishimaru, Yasushi Tanaka, Michiko Yamashita, Mika Sakaki, Yoshio Hayashi and Toshiaki Sano : Unique cell membrane expression if topoisomerase-ll alpha as a useful diagnostic marker of liposarcoma, Pathology International, Vol.54, No.3, 145-150, 2004.
(要約)
Topoisomerase-II alpha (Topo-II alpha) is known as a cell cycle-related intranuclear marker. To the best of our knowledge, the expression of Topo-II alpha on extranuclear sites has not been reported. The aim of the present study was to determine the usefulness of Topo-II alpha immunostaining for detecting the lipoblasts that are essential to diagnosing liposarcoma. Surgical specimens, including benign lipomatous tumors (four cases), well-differentiated liposarcomas (three cases), myxoid liposarcomas (six cases), pleomorphic liposarcomas (two cases), dedifferentiated liposarcomas (two cases), myxoid malignant fibrous histiocytomas (six cases), and one case of mesenteric panniculitis, were studied. Samples were immunostained using antibodies for Topo-II alpha, S-100 protein and Ki-67. In addition, we used the western blot method to investigate immunohistochemical-affinity in adipocytes. Mature adipocytes and lipoblasts in all of the benign and malignant lipomatous tumors intensively expressed cell contours positivity for Topo-II alpha. Cytoplasm of the lipoblasts occasionally reacted to the antibody and highlighted intracytoplasmic small unilocular, multivacuolated, or bubble-like patterns. Western blot analysis confirmed a 70 kDa product reactive to Topo-II alpha in the cell membrane fragment of mature adipocytes. S-100 protein expressed adipocytes and lipoblasts, but the detection of lipoblasts was not as easy as in Topo-II alpha immunostaining. Immunoreactivity of Ki-67 was limited to the nuclei, and the nuclear labeling index of Ki-67 correlated with that of Topo-II alpha. The immunoreactivity of Topo-II alpha for lipoblasts was more sensitive and obvious than those of S-100 protein. Immunostaining using the antibody for Topo-II alpha seems to be useful in recognizing lipoblasts that have been overlooked in hematoxylin-eosin-stained preparations, and is a useful marker for diagnosing liposarcoma.
Naozumi Ishimaru, Rieko Arakaki, Nobuhiko Katunuma and Yoshio Hayashi : Critical Role of Cathepsin-Inhibitors for Autoantigen Processing and Autoimmunity, Advances in Enzyme Regulation, Vol.44, No.1, 309-320, 2004.
Rieko Arakaki, Naozumi Ishimaru, Ichiro Saito, Masaru Kobayashi, Natsuo Yasui, Takayuki Sumida and Yoshio Hayashi : Development of Autoimmune Exocrinopathy Resembling Sjogren's Syndrome in Adoptively Transferred Mice With Autoreactive CD4+ T Cells, Arthritis and Rheumatism, Vol.48, No.12, 3603-3609, 2003.
(要約)
The pathologic mechanisms responsible for organ-specific tissue damage in primary Sjögren's syndrome (SS) remain unclear, but it has been suggested that the pathology is mediated by autoreactive CD4+ T cells infiltrating the salivary and lacrimal glands. This study was undertaken to investigate whether alpha-fodrin autoantigen-specific autoreactive CD4+ T cells are capable of inducing autoimmune lesions. A total of 45 synthetic alpha-fodrin peptides designed to be 20 amino acid residues in length were generated. To establish an autoreactive T cell line, limiting dilution analysis (LDA) was performed on lymph node cells (LNCs) in the presence of alpha-fodrin peptides. The effects of adoptive transfer of autoreactive CD4+ T cells into normal syngeneic recipients were investigated. Autoreactive CD4+ T cell lines that recognize synthetic alpha-fodrin peptide, which produced Th1 cytokines and showed cytotoxic activities, were established in a murine model for SS. T cell receptor V(beta) usage and third complementarity-determining region (CDR3) sequences indicated that in some cases V(beta)6-CDR3 genes matched between the tissue-infiltrating T cells and the autoreactive T cell lines. Adoptive transfer of the autoreactive CD4+ T cells into normal syngeneic recipients induced autoimmune lesions quite similar to those of SS. Our data help to elucidate the pathogenic mechanisms responsible for tissue destruction in autoimmune exocrinopathy and indicate that autoreactive CD4+ T cells play a pivotal role in the development of murine SS.
(キーワード)
Adoptive Transfer / Amino Acid Sequence / Animals / Base Sequence / CD4-Positive T-Lymphocytes / Carrier Proteins / Complementarity Determining Regions / Disease Models, Animal / Mice / Mice, Mutant Strains / Microfilament Proteins / Molecular Sequence Data / Sjogren's Syndrome / Specific Pathogen-Free Organisms
Moyamoya disease (MMD) is a rare entity that results in progressive occlusion of the arteries of the circle of Willis, but the pathogenesis of MMD is unknown. MMD sera (n=32) were tested for anti-endothelial cell antibodies by enzyme-linked immunoassays and flow cytometric analysis. Apoptosis was induced in human umbilical vein endothelial cells by tumor necrosis factor-alpha. We found that a high proportion of MMD sera had anti-endothelial cell antibodies with apoptotic stimuli. Prominent reactivities of MMD sera (72%) with recombinant human alpha-fodrin were observed. Our study demonstrates that MMD sera contain a high incidence of anti-alpha-fodrin autoantibodies, providing new insight into the mechanisms of occlusion of MMD arteries.
Naozumi Ishimaru, Rieko Arakaki, Megumi Watanabe, Kobayashi Masaru, Katsushi Miyazaki and Yoshio Hayashi : Development of autoimmune exocrinopathy resembling Sjogren's syndorome in estorogen deficient mice of healthy background, The American Journal of Pathology, Vol.163, No.4, 1481-1490, 2003.
(要約)
Although a number of autoimmune diseases are known to develop in postmenopausal women, the mechanisms by which estrogen deficiency influences autoimmune lesions remain unclear. We speculate that antiestrogenic actions might be a potent factor in the formation of pathogenic autoantigens. Previously, we have identified 120-kd alpha-fodrin as an important autoantigen in Sjögren's syndrome (SS). When healthy C57BL/6 (B6) mice were treated with an ovariectomy (Ovx), we found a significant increase in TUNEL(+)-apoptotic epithelial cells in the salivary gland cells associated with alpha-fodrin cleavage during 2 and 3 weeks after Ovx. By contrast, no apoptotic cells were found in estrogen receptor-alpha knockout mice. In in vitro studies using primary cultured mouse salivary gland cells and human salivary gland cells, we found a cleavage product of 120-kd alpha-fodrin in cells that had undergone tamoxifen (Tam)-induced apoptosis through caspase activation, especially caspase-1. Adoptive transfer of alpha-fodrin-reactive T cells into Ovx-B6 and -SCID mice resulted in the development of autoimmune exocrinopathy quite similar to SS. These results suggest that estrogen deficiency exerts a crucial influence on autoantigen cleavage, and may cause, in part, autoimmune exocrinopathy in postmenopausal women.
Although synovial chondromatosis occurs frequently near the ends of long bones, such as the knee, elbow, and hip joints, it rarely affects the temporomandibular joint.<BR>A 52-year-old woman visited our hospital because of swelling in the left side of the temporomandibular joint and trismus. On magnetic resonance imaging, the lesion was found to be chondroid tissue between the left mandibular fossa and the condylar head. Magnetic resonance imaging revealed a high signal mass measuring 35×40mm around the left condylar head. The clinical diagnosis was synovial chondromatosis. Synovectomy and removal of the mass were performed with the patient under general anesthesia. Histopathologically, the mass was formed by typical chondrocytes and was diagnosed to be synovial chondromatosis. The patient has been followed up for 1 year 4 months. There has been no reccurence of the mass or temporomandibular joint disorder.
Nobuhiko Katunuma, Yoishi Matsunaga, Kunisuke Himeno and Yoshio Hayashi : Insights into the roles of cathepsins in antigen processing and presentation revealed by specific inhibitors, Biological Chemistry, Vol.384, No.6, 883-890, 2003.
(要約)
Eleven human cathepsins have been identified, however, the in vivo roles of individual cathepsins are still largely unknown. In this brief review we will summarize the functions of individual cathepsins in antigen processing and presentation, which are the initial steps of the immune response. Two general inhibitors of papain-like cysteine proteases, E-64 and pyridoxal phosphate, can completely suppress antigen presentation in vivo. To evaluate the contribution of individual cathepsins, specific inhibitors have been developed based on cathepsin tertiary structures: CA-074 for cathepsin B, CLIK-148 and -195 for cathepsin L, CLIK-60 for cathepsin S. Administration of CA-074, a cathepsin B inhibitor, suppresses the response to exogenous antigens, such as hepatitis B virus antigen, ovalbumin and Leishmania major antigen, and induces switching of the helper T cell responses from Th-2 to Th-1 of CD4+ T cells, thereby downregulating the production of IgE and IgG1. Administration of the cathepsin S inhibitor CLIK-60 impairs presentation of an autoantigen, alpha-fodrin, in Sjogren's syndrome and suppresses the Th-1 response and autoantibody production.
Yoshifumi Nagata, Hiroko Inoue, Koichi Yamada, Hiroyuki Higashiyama, Kenji Mishima, Yasuhiro Kizu, Ienaka Takeda, Fumio Mizuno, Yoshio Hayashi and Ichiro Saito : Activation of Epstein-Barr virus by saliva from Sjogren's syndrome patients, Immunology, Vol.111, No.2, 223-229, 2003.
(要約)
The aim of this study was to examine the mechanism of Epstein-Barr virus (EBV) activation by soluble factors from the inflamed salivary glands of patients with Sjogren's syndrome (SS). Saliva from SS patients was used to examine the regulation of EBV activation by an inflammatory salivary microenvironment. Transient transfection of the EBV-negative salivary gland cell line (HSY) with BZLF1, a trans-activating EBV gene promoter-fusion construct (Zp-luc), was used in this study. The results showed that under conditions where the BZLF1 promoter is activated by potent stimuli, SS saliva (from eight of 12 patients) exerts a significant effect on expression of the luciferase gene. A specific inhibitor of protein kinase C did not affect the SS saliva-induced Zp-luc activity, whereas treatment with inhibitors of calmodulin, calcineurin and IP3, dose-dependently decreased this induction. Transforming growth factor beta1 (TGF-beta1), which is known to be expressed in SS salivary glands, dose-dependently induced Zp-luc activity. Hence, these results demonstrate the activation of EBV by SS saliva and suggest that EBV activation at the inflammatory site may occur in the presence of TGF-beta1 via triggering of the mitogen-activated protein kinase (MAPK) kinase signalling pathway.
(キーワード)
Adult / Biological Factors / Calcium-Calmodulin-Dependent Protein Kinases / Cell Line / DNA-Binding Proteins / Enzyme Inhibitors / Female / Herpesvirus 4, Human / Humans / MAP Kinase Signaling System / Promoter Regions, Genetic / Protein Kinase C / Saliva / Salivary Glands / Signal Transduction / Sjogren's Syndrome / Trans-Activators / Transfection / Transforming Growth Factor beta / Transforming Growth Factor beta1 / Viral Proteins / Virus Activation
Kaoru Saegusa, Naozumi Ishimaru, Kumiko Yanagi, Rieko Arakaki, Kouichi Ogawa, Ichiro Saito, Nobuhiko Katunuma and Yoshio Hayashi : Cathepsin S-inhibitor prevents autoantigen presentation and autoimmunity, The Journal of Clinical Investigation, Vol.110, No.3, 361-369, 2002.
(要約)
The cysteine endoprotease cathepsin S mediates degradation of the MHC class II invariant chain Ii in human and mouse antigen-presenting cells. Studies described here examine the functional significance of cathepsin S inhibition on autoantigen presentation and organ-specific autoimmune diseases in a murine model for Sjögren syndrome. Specific inhibitor of cathepsin S (Clik60) in vitro markedly impaired presentation of an organ-specific autoantigen, 120-kDa alpha-fodrin, by interfering with MHC class II-peptide binding. Autoantigen-specific T cell responses were significantly and dose-dependently inhibited by incubation with Clik60, but not with inhibitor s of cathepsin B or L. Clik60 treatment of mouse salivary gland cells selectively inhibited autopeptide-bound class II molecules. Moreover, the treatment with Clik60 in vivo profoundly blocked lymphocytic infiltration into the salivary and lacrimal glands, abrogated a rise in serum autoantibody production, and led to recovery from autoimmune manifestations. Thus, inhibition of cathepsin S in vivo alters autoantigen presentation and development of organ-specific autoimmunity. These data identify selective inhibition of cysteine protease cathepsin S as a potential therapeutic strategy for autoimmune disease processes.
Kenji Mishima, Kazuya Inoue and Yoshio Hayashi : Overexpression of extracelular-signal regulated kinases on oral squamous cell carcinoma, Oral Oncology, Vol.38, No.5, 468-474, 2002.
(要約)
Mitogen-activated protein kinases (MAPKs) consist of major three subfamilies, extracellular-signal regulated kinases (ERK MAPKs), the c-Jun N-terminal kinases/stress activated protein kinases (JNK MAPKs/SAP MAPKs), and p38 MAPKs. ERK MAPKs pathway is one of the most important pathways for cell proliferation. ERK MAPKs are located at downstream of a lot of growth factors (epidermal growth factor (EGF), nerve growth factor (NGF), platelet-derived growth factor (PDGF), etc.), the overexpressions and activation of which are frequently detected on a number of cancers including oral squamous cell carcinoma (OSCC). These data indicate that overexpression and activation of ERK MAPKs play an important role in cancer progression. On the contrary, JNK MAPKs are possible regulators of cell death induced by chemotherapeutic agents. p38 MAPKs are activated by pro-inflammatory cytokines and inflammatory drugs (non-steroidal anti-inflammatory drug), which are known to suppress cancer growth. These findings imply that each MAPKs can be molecular targets for cancer therapy in OSCC and its investigation is very important things in OSCC.
Masayuki Azuma, Keiko Aota, Tetsuya Tamatani, Katsumi Motegi, Yuki Ashida, Yoshio Hayashi, Mitsunobu Sato and Mitsunobu Sato : Suppression of TNF-α-induced MMP 9 production in human salivary gland acinar cells by cepharanthine occurs via down-regulation of NFκB, --- A possible therapeutic agent for preventing the destruction of the acinar structure in the salivary glands of Sjögren's syndrome patients ---, Arthritis and Rheumatism, Vol.46, No.6, 1585-1594, 2002.
(要約)
Our previous results suggested that suppression of tumor necrosis factor alpha (TNFalpha)-induced matrix metalloproteinase 9 (MMP-9) could prevent the destruction of acinar tissue in the salivary glands of patients with Sjögren's syndrome (SS). The present study was undertaken to investigate the effect of cepharanthine on the suppression of TNFalpha-induced MMP-9 production in NS-SV-AC, an SV40-immortalized normal human acinar cell clone. After pretreatment with or without cepharanthine, NS-SV-AC cells were treated with TNFalpha alone or with a combination of TNFalpha and cepharanthine. The expression of MMP-9 was then examined at the protein and messenger RNA levels. In addition, the effect of cepharanthine on the morphogenetic behavior of NS-SV-AC cells cultured on type IV collagen-coated dishes in the presence of TNFalpha was examined. Although TNFalpha induced the production of MMP-9 in NS-SV-AC cells, this production was greatly suppressed when cells were pretreated with cepharanthine, followed by treatment with both TNFalpha and cepharanthine. In addition, cepharanthine suppressed the TNFalpha-stimulated NF-kappaB activity by partly preventing the degradation of IkappaBalpha protein in NS-SV-AC cells. When NS-SV-AC cells were seeded on type IV collagen-coated dishes in the presence of both TNFalpha and plasmin, type IV collagen interaction with the cells was lost and the cells entered apoptosis. However, pretreatment with cepharanthine restored the aberrant in vitro morphogenesis of the NS-SV-AC cells. These results may indicate a molecular mechanism by which cepharanthine is able to protect against the destruction of the acinar structure in salivary glands from patients with SS.
Kaoru Saegusa, Naozumi Ishimaru, Kumiko Yanagi, Kenji Mishima, Rieko Arakaki, Takashi Suda, Ichiro Saito and Yoshio Hayashi : Prevention and induction of autoimmune exocrinopathy is dependent on pathogenic autoantigen cleavage in murine Sjogren's syndrome, The Journal of Immunology, Vol.169, No.2, 1050-1057, 2002.
(要約)
The in vivo role of autoantigen cleavage during apoptosis in autoimmune diseases remains unclear. Previously, we found a cleavage product of 120-kDa alpha-fodrin as an important autoantigen in the pathogenesis of primary Sjögren's syndrome (SS). In the murine primary SS model, tissue-infiltrating CD4(+) T cells purified from the salivary glands bear a large proportion of Fas ligand, and the salivary gland duct cells constitutively possess Fas. Infiltrating CD4(+) T cells, but not CD8(+) T cells, identified significant (51)Cr release against mouse salivary gland cells. In vitro studies demonstrated that apoptotic mouse salivary gland cells result in a specific alpha-fodrin cleavage into 120 kDa and that preincubation with caspase inhibitor peptides blocked alpha-fodrin cleavage. In vivo treatment with caspase inhibitors N-benzyloxycarbonyl-Val-Ala-Asp fluoromethyl ketone and N-acetyl-Asp-Glu-Val-Asp-al-CHO into the murine model results in dramatic inhibitory effects on the development of autoimmune lesions and in restoration of sicca syndrome. Furthermore, we found that immunization with recombinant alpha-fodrin protein identical with an autoantigen into normal recipients induced autoimmune lesions similar to SS. These data indicate that prevention and induction of autoimmune exocrinopathy is dependent on autoantigen cleavage via caspase cascade and that caspase inhibitors might provide a new therapeutic option directed at reducing tissue damage in the murine model for SS.
Naozumi Ishimaru, Kumiko Yanagi, Kouichi Ogawa, Takashi Suda, Ichiro Saito and Yoshio Hayashi : Possible Role of Organ-Specific Autoantigen for Fas Ligand-Mediated Activation-Induced Cell Death in Murine Sjögren's Syndrome1, The Journal of Immunology, Vol.167, No.10, 6031-6037, 2001.
(要約)
Activation-induced cell death (AICD) is a well-known mechanism of peripheral T cell tolerance that depends upon an interaction between Fas and Fas ligand (FasL). In this study, we demonstrate that the administration of a soluble form of anti-FasL Ab, FLIM58, results in severe destructive autoimmune exocrinopathy in the murine model of human Sjögren's syndrome (SS), and we found that an organ-specific autoantigen may play an important role on down-modulation of AICD. A high titer of serum autoantibodies against 120-kDa alpha-fodrin autoantigen was detected in the FLIM58-treated mice, and splenic T cell culture supernatants contained high levels of IFN-gamma. In vitro T cell apoptosis assay indicated that FasL-mediated AICD is down-regulated by autoantigen stimulation in spleen cells from the murine SS model, but not from Fas-deficient MRL/lpr mice and FasL-deficient MRL/gld mice. FasL undergo metalloproteinase-mediated proteolytic processing in their extracellular domains, resulting in the release of soluble trimeric ligands (soluble FasL). We showed that the processing of soluble FasL occurs in autoantigen-specific CD4(+) T cells, and that a significant increase in expressions of metalloproteinase-9 mRNA was observed in spleen cells from SS model mice. These findings indicate that the increased generation of soluble FasL inhibits the normal AICD process, leading to the proliferation of effector CD4(+) T cells in the murine SS model.
Hiroko Inoue, Kazuo Tsubota, Masafumi Ono, Yasuhiro Kizu, Fumio Mizuno, Kenzo Takada, Koichi Yamada, Kumiko Yanagi, Yoshio Hayashi and Ichiro Saito : Possible Involvement of EBV-Mediated α-Fodrin Cleavage for Organ-Specific Autoantigen in Sjogren's Syndrome, The Journal of Immunology, Vol.166, No.9, 5801-5809, 2001.
(要約)
A cleavage product of alpha-fodrin may be an important organ-specific autoantigen in the pathogenesis of Sjogren's syndrome (SS), but the mechanisms of alpha-fodrin cleavage remain unclear. Since EBV has been implicated in the pathogenesis of SS, we determined whether EBV activation could induce the SS-specific 120-kDa autoantigen alpha-fodrin. ZEBRA mRNA expression, a marker for activation of the lytic cycle of EBV, was found in the salivary gland tissues from SS patients, but not in those from control individuals. ZEBRA-expressing lymphoid cells were also found in the SS glands in double-stained immunohistochemistry. Furthermore, a significant link between production of Abs against 120-kDa alpha-fodrin and reactivated EBV Ag was found in sera from patients with SS, but not in those from control individuals. EBV-activated lymphoid cells showed specific alpha-fodrin cleavage to the expected 120-kDa fragments in vitro. Pretreatment with caspase inhibitors inhibited cleavage of alpha-fodrin. Thus, an increase in apoptotic protease activities induced by EBV reactivation may be involved in the progression of alpha-fodrin proteolysis in the development of SS.
Nobuaki Maeno, Syuji Takei, Hiroyuki Imanaka, Hiroshi Oda, Yoshio Hayashi and Koichiro Miyata : Anti-alpha-fodrin antibodies in Sjogren's syndrome in children, The Journal of Rheumatology, Vol.28, No.4, 860-864, 2001.
(要約)
To investigate the prevalence of anti-alpha-fodrin antibody specific for adult Sjögren's syndrome (SS) in patients with juvenile onset SS. Serum anti-alpha-fodrin antibody was examined in 15 patients with juvenile SS (11 cases of primary SS and 4 secondary SS) and in 16 children with systemic lupus erythematosus (SLE) by Western blot analysis using a recombinant 120 kDa alpha-fodrin fusion protein. All the 15 serum samples from patients with SS reacted with a recombinant alpha-fodrin fusion protein in Western blot analysis. In contrast, reactivity was found in only 2 of the 16 patients with SLE. The clinical features of the 15 patients with juvenile onset SS were very specific; only 4 patients complained of dryness, while 6 had abnormal excretion ability. Salivary gland enlargement was the most common clinical manifestation. Characteristic laboratory findings in juvenile onset SS included a higher prevalence of antinuclear antibodies, anti-SSA/Ro antibodies, and rheumatoid factor, as well as increased erythrocyte sedimentation rate and hypergammaglobulinemia. The pathogenesis of juvenile SS seems to be the same as that of adult SS, although subjective symptoms of dryness are less frequent in juvenile cases. This discrepancy may indicate that SS is a slowly progressive disease with a long time span. The anti-alpha-fodrin antibody is likely to be a reliable diagnostic marker for juvenile SS.
Ichiro Kobayashi, Nobuaki Kawamura, Motohiko Okano, Takaaki Shikano, Masahiko Mizutomo, Yoshio Hayashi and Kunihiko Kobayashi : Anti-α-Fordrin Autoantibody Is an Early Diagnostic Marker for Childhood Primary Shogren Syndrome, The Journal of Rheumatology, Vol.28, No.2, 363-365, 2001.
(要約)
alpha-fodrin is a recently identified autoantigen associated with adult primary Sjögren's syndrome (SS). We tested whether anti-alpha-fodrin antibody could also be used as a diagnostic marker for childhood SS. We performed immunoblot analysis of sera from 7 patients with childhood primary SS using glutathione-S-transferase alpha-fodrin fusion protein as an antigen. Anti-alpha-fodrin antibody was detected in sera from all 7 patients with childhood primary SS, 2 of 4 with secondary SS, and one of 7 with systemic lupus erythematosus, but in no other healthy controls. The anti-alpha-fodrin autoantibody was detected before anti-SSA or SSB antibody became positive; thus anti-alpha-fodrin antibody could be a useful marker for the early diagnosis of SS.
(キーワード)
α-Fordrin / Autoantibody / Childhoood Sjogren Symdrome / Early Diagnosis
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 11246678
Kazuo Tsubota, Hiromi Fujita, Kimie Tadano, Tsutomu Takeuchi, Tadahiro Murakami, Ichiro Saito and Yoshio Hayashi : Improvement of Lacrimal Function by Topical Application of CyA in Murine Models of Sjögren's Syndrome, Investigative Ophthalmology & Visual Science, Vol.42, No.1, 101-110, 2001.
(要約)
The object of this study was to evaluate improvement of lacrimal gland (LG) function after topical cyclosporin A (CyA). Topical CyA (0.01% and 0.1%) was applied to two mouse models of Sjögren's syndrome, the NFS/sld after thymectomy and the nonobese diabetic (NOD) mouse, and the functional integrity of the lacrimal gland was evaluated by measuring basal and stimulated tear secretion and its histologic integrity by examining it for apoptosis and lymphocyte invasion. After treatment with CyA at 0.1% in the NFS/sld mice, tear function increased, and there was a decrease in lymphocyte infiltration of the LG and a decrease in apoptotic figures among the acinar cells. In the NOD mice, tear function also improved, but there was no associated decrease in lymphocyte infiltration. However, the expression of Fas ligand (FasL) in NOD mice by infiltrating lymphocytes was suppressed with 0. 1% CyA eye drops. CyA appears to improve tear secretion in mouse models of Sjögren's syndrome by preventing lymphocyte-induced apoptosis of acinar cells. In one model this was achieved by preventing lymphocyte infiltration and in the other by reducing expression of FasL expression on infiltrating lymphocytes.
Kaoru Saegusa, Naozumi Ishimaru, Norio Haneji, Kumiko Yanagi, Tomoko Yoneda, Ichiro Saito and Yoshio Hayashi : Mechanisms of neonatal tolerance induced in an animal model for primary Sjogren's syndrome by intravenous administration of autoantigen, Scandinavian Journal of Immunology, Vol.52, No.3, 264-270, 2000.
(要約)
Neonatal exposure to autoantigen is believed to induce effective antigen-specific T-cell tolerance in experimental models of autoimmunity. We have identified 120 kDa alpha-fodrin autoantigen in an animal model for primary Sjögren's syndrome (SS), that has been determined as a candidate autoantigen in both an animal model and the patients with primary SS. We demonstrate here that neonatal injection of autoantigen induce relevant tolerance when treated with intravenous (i.v.) administration within 24 h after birth, but not with i.v. injection after the thymectomy or with intraperitoneal injection. Autoantigen-specific T-cell response was significantly reduced in mice induced neonatal tolerance, and the activation markers of splenic CD4+ T cells were down-regulated in mice treated with neonatal administration. Because we detected that neonatal i.v. injection of autoantigen prevented Th1 response, it is possible that the autoantigen administration within 24 h after birth induce regulatory T cells that had a protective effect against Th1-mediated autoimmune diseases. These results indicate that the prevention of the spontaneous anti-120 kDa alpha-fodrin response in vivo, by tolerization of the autoantigen-reactive T cells, blocked the development of autoimmune lesions in an animal model for primary SS.
Hideaki Nakamura, Atsushi Kawakami, Satoshi Yamasaki, Tomoki Nakashima, Makoto Kamachi, Kiyoshi Migita, Yojiro Kawabe, Tatsufumi Nakamura, Takehiko Koji, Yoshio Hayashi and Katsumi Eguchi : Expression and Function of X Chromosome-Linked Inhibitor of Apoptosis Protein in Sjogren's Syndrome, Laboratory Investigation; a Journal of Technical Methods and Pathology, Vol.80, No.9, 1421-1427, 2000.
(要約)
Apoptotic cell death in acinar and ductal epithelial cells is thought to play an important role in the development of salivary gland dysfunction in patients with Sjogren's syndrome (SS). We examined the expression of anti-apoptotic molecules in salivary glands from patients with SS. The labial salivary glands from six human T-cell leukemia virus (HTLV)-I-seronegative and eleven HTLV-I-seropositive SS patients were analyzed by immunohistochemistry. In vitro experiments were performed with a human salivary gland cell line (HSG cells). Immunohistologic analyses revealed that Bcl-2 and Bcl-x were preferentially expressed in salivary infiltrating mononuclear cells more than acinar and ductal epithelial cells. In contrast, strong X chromosome-linked inhibitor of apoptosis protein (XIAP) expression was evident in both acinar and ductal epithelial cells. The pattern of expression of these anti-apoptotic molecules was similar in both HTLV-I-seropositive and HTLV-I -seronegative SS patients. Western blot analysis confirmed expression of XIAP in cultured HSG cells. The expression of XIAP in HSG cells was increased by IL-1beta, TGF-beta1, or IL-10. However, XIAP expression was down-regulated by TNF-alpha, which induced apoptotic cell death of HSG cells with an increase in caspase-3 activity. These effects of TNF-alpha in HSG cells were antagonized by IL-1beta, TGF-beta1, or IL-10. Our results suggest that XIAP is important in regulating apoptotic cell death of acinar and ductal epithelial cells in patients with SS.
(キーワード)
Adult / Aged / Aged, 80 and over / Apoptosis / Female / Humans / Immunohistochemistry / Middle Aged / Proteins / Proto-Oncogene Proteins c-bcl-2 / Salivary Glands / Sjogren's Syndrome / Tumor Necrosis Factor-alpha / X-Linked Inhibitor of Apoptosis Protein / bcl-X Protein
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 11005210
Kaoru Saegusa, Naozumi Ishimaru, Kumiko Yanagi, Norio Haneji, Mizuho Nishino, Masayuki Azuma, Ichiro Saito and Yoshio Hayashi : Autoantigen-specific CD4+CD28low T cell subset prevents autoimmune exocrinopathy in murine Sjogren's syndrome, The Journal of Immunology, Vol.165, No.4, 2251-2257, 2000.
(要約)
Organ-specific autoimmune exocrinopathy resembling Sjögren's syndrome (SS) that spontaneously develops in NFS/sld mutant mice thymectomized 3 day after birth is dependent on Th1-type CD4+ T cells. We previously reported that a cleavage product of 120-kDa alpha-fodrin may be an important autoantigen in the pathogenesis of SS in both an animal model and the patients. We demonstrate that in an animal model of SS with overt exocrinopathy, a unique CD4+ T cell subset expressing CD28low is dramatically increased in spleen cells before the disease onset, but that the CD4+ T cells of diseased mice were virtually all CD28high. We found that the spleen cells in these mice before the disease onset showed a significant increase in autoantigen-specific T cell proliferation. Analysis of in vitro cytokine production by spleen cells indicated, before the disease onset, severely impaired production of IL-2 and IFN-gamma in the animal model, whereas high levels of IL-4 were observed. Expression of cytokine genes, including IL-4, IL-10, and TGF-beta, was detected in FACS-sorted CD4+CD28low T cells by RT-PCR analysis. Transfer of CD4+CD28low T cells into the animal model actually prevented the development of autoimmune lesions including autoantibody production. These results suggest that a CD4+CD28low T cell subset that is continuously activated by an organ-specific autoantigen may play a regulatory role in the development of organ-specific autoimmune disease in an animal model of SS.
Masayuki Azuma, Tetsuya Tamatani, Katsumi Motegi, Keiko Aota, Tsuyoshi Yamashita, Kouji Harada, Yoshio Hayashi and Mitsunobu Sato : Suppression of tumor necrosis factor-a-induced matrix metalloproteinase-9-production by introduction of a super-repressor form of IkB-cDNA into immotalized human salivary gland acinar cells, Arthritis and Rheumatism, Vol.43, No.8, 1756-1767, 2000.
(要約)
We have previously shown that specific enhancement in acinar cells of proteolytic activity induced by tumor necrosis factor alpha (TNFalpha) may be responsible for the destruction of the acinar structure in the salivary glands of patients with Sjögren's syndrome. Because matrix metalloproteinase 9 (MMP-9) is regulated by nuclear factor kappaB (NF-kappaB), we investigated the effect of a super-repressor form of inhibitor of nuclear factor kappaBalpha (srIkappaBalpha) on the suppression of TNFalpha-induced MMP-9 production in acinar cells. Two srIkappaBalpha complementary DNA (cDNA)-transfected acinar cell clones (ACMT-6 and ACMT-7) and 1 empty vector-transfected cell clone (ACpRc-1) were established. After treatment of cell clones with TNFalpha, the expression of MMP-9 was examined. In addition, the effect of TNFalpha on cell growth and the morphogenetic behavior of cell clones cultured on type IV collagen-coated dishes were examined. TNFalpha induced the production of MMP-9 in the ACpRc-1 cell clone, but greatly suppressed MMP-9 production in ACMT-6 and ACMT-7 clones. No apparent cytotoxic effect of TNFalpha treatment was observed in these cell clones. When ACpRc-1 cells were seeded on type IV collagen-coated dishes in the presence of both TNFalpha and plasmin, type IV collagen interaction with the cells was lost and the cells entered apoptosis. However, even when ACMT-6 and ACMT-7 cells were cultured under the same culture conditions as those for ACpRc-1, these cell clones attached to the substrate and grew consistently without showing apoptosis. Conclusion. These observations indicate that suppression of TNFalpha-induced MMP-9 production by the introduction of srIkappaBalpha cDNA corrected the aberrant in vitro morphogenesis of acinar cells grown on type IV collagen.
Keiko Aota, Masayuki Azuma, Tsuyoshi Yamashita, Testuya Tamatani, Katsumi Motegi, Naozumi Ishimaru, Yoshio Hayashi and Mitsunobu Sato : 5-Fluorouracil Induces Apoptosis through the Suppression of NF-κB Activity in Human Salivary Gland Cancer Cells, Biochemical and Biophysical Research Communications, Vol.273, No.3, 1168-1174, 2000.
(要約)
Activation of the transcription factor NF-kappaB results in protection against apoptosis, and the chemotherapeutic agent 5-Fluorouracil (5-FU) exerts its cytotoxic effect through the induction of apoptosis. Thus, we examined whether 5-FU could induce apoptosis through the suppression of NF-kappaB activity. We found that upon treatment of a human salivary gland cancer cell line (cl-1) with 5-FU, the NF-kappaB activity was suppressed in a time-dependent manner. This inhibition was mediated by a prevention of the degradation of the inhibitory IkappaB-alpha protein. In addition, the expression of TRAF-2 and cIAP-1, which are transcriptionally regulated by NF-kappaB and function as anti-apoptotic molecules through the interruption of caspase pathway, was also inhibited by 5-FU. Finally, the activity of caspase-8 and caspase-3 showed a significant increase in response to 5-FU. By flow cytometric analysis, 5-FU did not affect the expression level of Fas on the cell surface. Thus, our results suggest that one of the molecular mechanisms involved in 5-FU-induced apoptosis in cl-1 cells may be due to the suppression of NF-kappaB activity, resulting in the activation of the pro-apoptotic pathway.
Maki Kondo, Takashi Yamaoka, Soichi Honda, Yoshihiro Miwa, Maki Moritani, Kastuhiko Yoshimoto, Yoshio Hayashi and Mitsuo Itakura : The Rate of Cell Growth Is Regulated by Purine Biosynthesis via ATP Production and G to Sphase Transition, The Journal of Biochemistry, Vol.128, No.1, 57-64, 2000.
(要約)
We recently showed that an increased supply of purine nucleotides increased the growth rate of cultured fibroblasts. To understand the mechanism of the growth rate regulation, CHO K1 (a wild type of Chinese hamster ovary fibroblast cell line) and CHO ade (-)A (a cell line deficient in amidophosphoribosyltransferase, a rate-limiting enzyme of the de novo pathway) were cultured under various conditions. Moreover, a defective de novo pathway in CHO ade (-)A cells was exogenously restored by 5-amino-4-imidazole-carboxamide riboside, a precursor of the de novo pathway. The following parameters were determined: the growth rate of CHO fibroblasts, the metabolic rate of the de novo pathway, the enzyme activities of amidophosphoribosyltransferase and hypoxanthine phosphoribosyltransferase, the content of intracellular nucleotides, and the duration of each cell-cycle phase. We concluded the following: (i) Purine de novo synthesis, rather than purine salvage synthesis or pyrimidine synthesis, limits the growth rate. (ii) Purine nucleotides are synthesized preferentially by the salvage pathway as long as hypoxanthine is available for energy conservation. (iii) The GTP content depends on the intracellular ATP content. (iv) Biosynthesis of purine nucleotides increases the growth rate mainly through ATP production and promotion of the G(1)/S transition.
Naozumi Ishimaru, Tomoko Yoneda, Kaoru Saegusa, Kumiko Yanagi, Norio Haneji, Keiji Moriyama, Ichiro Saito and Yoshio Hayashi : Severe destructive autoimmune lesions with aging in murine Sjogren's syndrome through Fas-mediated apoptosis, The American Journal of Pathology, Vol.156, No.5, 1557-1564, 2000.
(要約)
When we evaluated the age-associated changes in autoimmune exocrinopathy in a NFS/sld murine model for primary Sjögren's syndrome (SS), severe destructive autoimmune lesions developed in the salivary and lacrimal glands in the aged mice, compared with those observed in the younger model. We detected a decreased secretion of saliva and tear flow in the aged group. A significant increase of TUNEL(+)-apoptotic epithelial duct cells in the salivary glands was detected in the aged SS animal model. A higher proportion of mouse salivary gland cells bearing Fas was found in the aged group, whereas no significant changes were seen on tissue-infiltrating CD4(+) T cells bearing FasL in the salivary glands from young and aged mice. We detected an increased cleavage product of organ-specific autoantigen, 120-kd alpha-fodrin, in the aged salivary gland tissues on immunoblotting, and an increase in serum autoantibody production against 120-kd alpha-fodrin by enzyme-linked immunosorbent assay. An increase in the proliferative response of splenic T cells against organ-specific autoantigen was observed, whereas nonspecific concanavalin A responsiveness was decreased in the aged mice. In addition, a decrease in Fas expression was found on splenic CD4(+) T cells in the aged mice, and anti-Fas mAb-stimulated apoptosis was down-regulated on CD4(+) T cells. These results indicate that age-associated dysregulation of CD4(+) T cells may play a crucial role on acceleration of organ-specific autoimmune lesions in a murine model for primary SS through Fas-mediated apoptosis.
Kaoru Saegusa, Naozumi Ishimaru, Kumiko Yanagai, Norio Haneji, Mizuho Nishino, Masayuki Azuma, Ichiro Saito and Yoshio Hayashi : Treatment with anti-CD 86 costimulatory molecule prevents the autoimmune lesions in murine Sjogren's syndrome(SS) through up-regulated Th2 response, Clinical and Experimental Immunology, Vol.119, 354-360, 2000.
82.
Maki Kondo, Takashi Yamaoka, Soichi Honda, Yoshihiro Miwa, Rumi Katashima, Maki Moritani, Katsuhiko Yoshimoto, Yoshio Hayashi and Mitsuo Itakura : The Rate of Cell Growth is Regulated by Purine Biosynthesis via ATP Production and G1 to S Phase Transition., The Journal of Biochemistry, Vol.128, No.1, 57-64, 2000.
Naozumi Ishimaru, Kaoru Saegusa, Kumiko Yanagi, Norio Haneji, Ichiro Saito and Yoshio Hayashi : Estrogen Deficiency Accelerates Autoimmune Exocrinopathy in Murine Sjögren's Syndrome through Fas-Mediated Apoptosis, The American Journal of Pathology, Vol.155, No.1, 173-181, 1999.
87.
Naozumi Ishimaru, Kaoru Saegusa, Kumiko Yanagi, Norio Haneji, Ichiro Saito and Yoshio Hayashi : Estrogen dediciency accelerates autoimmune exocrinopathy in murine Sjogren's syndrome through Fas-dediated apoptosis, The American Journal of Pathology, Vol.155, No.1, 173-181, 1999.
(要約)
Estrogenic action has been suggested to be responsible for the strong female preponderance of autoimmune diseases, but the role of estrogens in the female has not been well characterized. We evaluated the effects of estrogen deficiency in a murine model for autoimmune exocrinopathy of Sjögren's syndrome (SS). Severe destructive autoimmune lesions developed in the salivary and lacrimal glands in estrogen-deficient mice, and these lesions were recovered by estrogen administration. We detected an intense estrogen receptor in splenic CD8(+) T cells compared with that in CD4(+) T cells, and concanavalin-A-stimulated blastogenesis of splenic CD8(+) T cells with estrogens was much higher than that of CD4(+) T cells. We found a significant increase in serum autoantibody production against the organ-specific autoantigen alpha-fodrin. Moreover, an increased proportion of TUNEL+ apoptotic epithelial duct cells was observed in estrogen-deficient mice. It was demonstrated that Fas-mediated apoptosis in cultured salivary gland cells was clearly inhibited by estrogens in vitro. These results indicate that dysfunction of regulatory T cells by estrogen deficiency may play a crucial role on acceleration of organ-specific autoimmune lesions, and estrogenic action further influences target epithelial cells through Fas-mediated apoptosis in a murine model for SS.
Mariko Takahashi, Naozumi Ishimaru, Kumiko Yanagi, Kaoru Saegusa, Norio Haneji, Hiroshi Shiota and Yoshio Hayashi : Requirement for splenic CD4+ T cells in the immune privilege of the anterior chamber of the eye, Clinical and Experimental Immunology, Vol.116, No.2, 231-237, 1999.
Takahiro Watanabe, Tetsuya Tsuchida, Naoko Kanda, Katsunori Mori and Yoshio Hayashi : Anti-α-fodrin antibodies in Sjogren's syndrome and lupus erythematosus, Arch Dermatol, Vol.135, No.5, 535-539, 1999.
90.
Ichiro Saito, Kumiko Haruta, Misa Shimuta, Hiroko Inoue, Hiroshi Sakurai, Koichi Yamada, Naozumi Ishimaru, Hiroyuki Higashiyama, Takayuki Sumida, Hiroshi Ishida, Takashi Suda, Tetsuo Noda, Yoshio Hayashi and Kazuo Tsubota : Fas ligand-mediated exocrinopathy resembling Sjogren's syndrome in mice transgenic for IL-10, The Journal of Immunology, Vol.162, No.5, 2488-2492, 1999.
(要約)
Although IL-10 has been implicated in the pathogenesis of several autoimmune diseases, the mechanisms by which this cytokine mediates inflammatory lesions remain to be elucidated. Exocrine gland destruction is an important early step in the development of Sjögren's syndrome. To better understand the role of IL-10 in Sjögren's syndrome, we made transgenic mice in which the mouse IL-10 gene was regulated by the human salivary amylase promoter. Transgenic expression of IL-10 induced apoptosis of glandular tissue destruction and lymphocyte infiltration consisting primarily of Fas-ligand (FasL)+ CD4+ T cells, as well as in vitro up-regulation of FasL expression on T cells. These data suggest that overexpression of IL-10 in the glands and their subsequent Fas/FasL-mediated bystander tissue destruction is a causal factor in the development of this disease.
Sachiko Miyagawa, Kumiko Yanagi, Akira Yoshioka, Kin-ichi Kidoguchi, Toshihiko Shirai and Yoshio Hayashi : Neonatal lupus erythematosus: Maternal IgG antibodies bind to a recombinant NH2-terminal fusion protein encoded by human α-fodrin cDNA, The Journal of Investigative Dermatology, Vol.111, No.6, 1189-1192, 1998.
(要約)
IgG antibodies to a cleavage product of alpha-fodrin (120 kDa alpha-fodrin) have recently been identified as organ-specific autoantibodies in primary Sjögren's syndrome. In this study, we examined seroreactivity of mothers and infants with neonatal lupus erythematosus (NLE) to a recombinant NH2-terminal protein (120 kDa alpha-fodrin) of human alpha-fodrin. Serum samples were collected during the perinatal period in seven pregnancies of five mothers delivering offspring with NLE. Anti-120 kDa alpha-fodrin antibodies were identified by immunoblotting in six of seven perinatal maternal sera of offspring with NLE: one of two congenital heart block offspring and all five offspring with cutaneous NLE. These antibodies were placentally transmitted to infants. One of the five mothers had primary Sjögren's syndrome, and four were asymptomatic. One asymptomatic mother did not demonstrate anti-120 kDa alpha-fodrin activity at the time of the first delivery of a congenital heart block infant, but was found to be positive at the time of subsequent delivery of a second child with cutaneous NLE. We propose that maternal antibodies to 120 kDa alpha-fodrin may be an additional serologic marker for the risk of NLE in anti-Ro/SS-A positive women.
Kazuo Tsubota, Ichiro Saito, Naozumi Ishimaru and Yoshio Hayashi : Use of topical cyclosporin A in a primary Sjogren's syndrome mouse model, Investigative Ophthalmology & Visual Science, Vol.39, No.9, 1551-1559, 1998.
(要約)
A new animal model, the NFS/sld mutant mouse, was used for primary Sjögren's syndrome to investigate the efficacy of topical and systemic cyclosporin A (CyA) in preventing inflammation of the exocrine glands. Cyclosporin A was applied topically (0.01% and 0.1%, three times a day) or administered orally (10 mg/kg and 100 mg/kg, once a day) to mice from 6 to 16 weeks of age, after which the mice were killed. Topical CyA reduced lacrimal gland and submandibular gland inflammation without causing pathologic changes in other organs. Flow cytometry showed that CD44 expression of CD4 T cells from the submandibular lymph nodes was downregulated, whereas that of Mel14+ was upregulated. Using a reverse transcription-polymerase chain reaction assay, we determined that topical CyA significantly decreased the expression of mRNA of IL-2 and T-cell receptor-constant beta-chain. Topical CyA may be clinically useful in reducing the lymphocyte infiltration of lacrimal glands associated with Sjögren's syndrome.
Kaoru Saegusa, Naozumi Ishimaru, Kumiko Yanagi, Norio Haneji, Mizuho Nishino and Yoshio Hayashi : Mechanisms of transfer into SCID mice reconstituted with tissue-infiltrating cells from murine model for Sjogren's syndrome, Oral Med Pathol, Vol.3, 67-74, 1998.
Masayuki Azuma, Katsumi Motegi, Keiko Aota, Yoshio Hayashi and Mitsunobu Sato : Role of cytokines in the destruction of acinar structure in Sjogren's syndrome salivary glands, Laboratory Investigation; a Journal of Technical Methods and Pathology, Vol.77, No.3, 269-280, 1997.
96.
Norio Haneji, Takanori Nakamura, Koji Takio, Kumiko Yanagi, Hiroyuki Higashiyama, Ichiro Saito, Sumihare Noji, Hiromu Sugino and Yoshio Hayashi : Identification of α-fodrin as a candidate autoantigen in primary sjögren's syndrome, Science, Vol.276, No.5312, 604-607, 1997.
Mariko Takahashi, Naozumi Ishimaru, Kumiko Yanagi, Norio Haneji, Ichiro Saito and Yoshio Hayashi : High incidence of autoimmune dacryoadenitis in male non-obese diabetic (NOD) mice depending on sex steroid, Clinical and Experimental Immunology, Vol.109, 555-561, 1997.
(キーワード)
Sjogren's syndrome / Lacrimal gland / 炎症性サイトカイン受容体の情報伝達 (Signal transduction of proinflammatory cytokine receptors) / sex steroid / NOD mice
99.
Masayuki Azuma, Katsumi Motegi, Keiko Aota, Yoshio Hayashi and Mitsunobu Sato : Role of cytokines of acinar structure in Sjogren's syndrome salivary glands, Laboratory Investigation; a Journal of Technical Methods and Pathology, Vol.77, No.3, 269-280, 1997.
100.
Kumiko Yanagi, Norio Haneji, Naozumi Ishimaru, Ichiro Saito and Yoshio Hayashi : Analysis of T cell receptor Vb usage in the autoimmune sialadenitis of non-obese diabetic (NOD) mice, Clinical and Experimental Immunology, Vol.110, 440-446, 1997.
Yoshio Hayashi, Norio Haneji, Hironori Hamano, Kumiko Yanagi, Mariko Takahashi and Naozumi Ishimaru : Effector Mechanism of Experimental Autoimmune Sialadenitis in the Mouse Model for Primary Sjogren's Syndrome, Cellular Immunology, Vol.171, No.2, 217-225, 1996.
(要約)
We have recently established a new animal model for primary Sjögren's syndrome in NFS/sld mutant mice thymectomized 3 days after birth (3dTX) bearing an autosomal recessive gene with sublingual gland differentiation arrest. In this study, we analyze developing mechanisms of experimental autoimmune sialadenitis (EAS) in the mouse model, focusing on local expressions of cytokine and cell adhesion molecule genes by reverse transcriptase-polymeric chain reaction (RT-PCR) and immunohistochemistry, kinetic analysis of splenic lymphocytes expressing activation markers, and I-Aq class-II molecules by flow cytometry (FACS). We found up-regulation of local cytokine genes (IL-1 beta, TNF-alpha, IL-2, IFN-gamma, IL-6, IL-10, IL-12p40) and cell adhesion molecule genes (ICAM-1, LFA-1, CD44, Mel-14) in the salivary glands from mice with EAS by RT-PCR, which were supported by immunohistochemistry. FACS analysis demonstrated that a significant proportion of splenic CD4+ T cells express activation markers (CD44, LFA-1, Mel-14low, CD45RB(low)) at a high level and an increase in expression of B220+ B cells bearing I-Aq class-II molecules. These data suggest that spontaneous EAS in 3dTX NFS/sld mutant mice may be triggered by an in situ activation of autoreactive CD4+ T cells comprising unique cytokine profile (high levels of IL-2, IFN-gamma, IL-10, and IL-12p40 mRNA) in the salivary glands.
Youji Miyamoto, Kenji Fujisawa, Masaru Nagayama, Norio Haneji and Yoshio Hayashi : Carcinoma in Pleomorphic Adenoma of the Tongue ; Report of a case, Oral Medicine & Pathology, Vol.1, No.2, 111-114, 1996.
(要約)
A carcinoma in pleomorphic adenoma occurring in the tongue of a 47-year-old Japanese woman is reported. The present report is the first to describe a carcinoma in pleomorphic adenoma derived from the posterior lingual glands.
Mariko Takahashi, Yasuo Mimura, Hironori Hamano, Norio Haneji, Kumiko Yanagi and Yoshio Hayashi : Mechanism of the development of autoimmune dacryoadenitis in the mouse model for primary Sjogren's syndrome, Cellular Immunology, Vol.170, No.1, 54-62, 1996.
(要約)
To elucidate the mechanism of development in autoimmune lacrimal gland disease, we analyzed different aspects of autoimmune dacryoadenitis in a newly established mouse model for primary Sjögren's syndrome, focusing on the local expressions of cytokine genes, and the repertoire of T cell receptor (TCR) V beta genes transcribed within the inflammatory infiltration in the lacrimal glands. We found that the vast majority of inflammatory infiltration into the lacrimal glands were CD4+ V beta 8+ T cells. We detected the up-regulation of local cytokine genes (IL-1 beta, TNF-alpha, IL-2, IFN-gamma, IL-10, IL-12p40) in the lacrimal glands with very early inflammatory lesions by reverse transcriptase (RT)-PCR analysis. The predominant expression of the V beta 8 gene segment was detected from a very early stage, while extensive age-related diversity of TCR V beta gene usage was observed. Single-strand conformation polymorphism (SSCP) analysis demonstrated a distinct and a common binding pattern in the PCR product of the V beta 8 gene on the infiltrating cells during the course of the disease. These data suggest that in autoimmune dacryoadenitis of the mouse model for primary Sjögren's syndrome there may be a restricted usage of TCR V beta elements on a very early stage of the autoimmune lesion to recognize unknown self-antigen, and the autoreactive CD4+ T cells constitute a unique cytokine profile in the autoimmune lacrimal gland disease.
Youji Miyamoto, Masaru Nagayama and Yoshio Hayashi : Verruciform xanthoma occurring within oral lichen planus, Journal of Oral Pathology & Medicine, Vol.25, No.4, 188-191, 1996.
105.
Kumiko Yanagi, Norio Haneji, Hironori Hamano, Mariko Takahashi, Hiroyuki Higashiyama and Yoshio Hayashi : In vivo role of IL-10 and IL-12 during development of Sjogren's syndrome in MRL/lpr mice, Cellular Immunology, Vol.168, No.2, 243-250, 1996.
(要約)
Expression of local cytokine genes including interleukin 10 (IL-10) and IL-12 was analyzed in the salivary gland tissues of MRL/lpr mice with Sjögren's syndrome. We demonstrate a significant role of IL-10 and IL-12 in vivo during development of Sjögren's syndrome in MRL/lpr mice. IL-10 mRNA expression was detected before the onset of disease and was upregulated during the course of autoimmune sialadenitis by RT-PCR. A predominant level of expression of IL-12 mRNA was also detected earlier in the proinflammatory stage of autoimmune sialadenities. Moreover, MHC class II (I-Ak) mRNA was detected before the onset of inflammatory lesions until older ages in the salivary glands of MRL/lpr mice. These results suggest that endogenous IL-10 and IL-12 may play important roles on immune-mediated destruction of the salivary glands during development of organ-specific autoimmunity in MRL/lpr mice.
Sachiko Iseki, Akihiro Araga, Hideyo Ohuchi, Tsutomu Nohno, Hidefumi Yoshioka, Yoshio Hayashi and Sumihare Noji : Sonic hedgehog is expressed in epithelial cells during development of whiskar, hair, and tooth, Biochemical and Biophysical Research Communications, Vol.218, No.3, 688-693, 1996.
Tooth development / Polarizing activity / Sonic hedgehog / Enamel knot
108.
Yoshio Hayashi, Norio Haneji and Hironori Hamano : Cytokine gene expression and autoantibody production in Sjogren's syndrome in MRL/lpr mice, Autoimmunity, Vol.23, No.4, 269-277, 1996.
(要約)
In an attempt to elucidate the mechanism of development of organ-specific autoimmune lesions resembling human Sjögren's syndrome of MRL/lpr mice, we have analyzed local cytokine gene expressions and organ-specific autoantibody production in vivo. We have demonstrated that a major proportion of T cells bearing CD4 and V(beta)8 molecules are essentially responsible for triggering the autoimmunity in the salivary glands of MRL/lpr mice. The local cytokine gene expressions including interferon(IFN)-gamma, IL-12(p40) mRNAs were observed during the course of murine Sjogren's syndrome in MRL/lpr autoimmune strain. In particular, a high level of local expressions of IL-12 mRNA was detected earlier in the proinflammatory stage of autoimmune lesions. A significant level of local expression of MHC class-II(I-Ak) mRNA was detected before the onset of inflammatory lesions in the salivary glands, and I-Ak-positive epithelial duct cells were frequently observed in the salivary glands of MRL/lpr mice. In addition, we found the salivary gland-specific autoantibody in sera from MRL/lpr mice with early phase of autoimmune lesions by immunoblot analysis. These results suggest that cytokine gene stimulation and autoantibody production are essentially involved in the development of organ-specific autoimmune lesions in Sjögren's syndrome of MRL/lpr mice.
Kumiko Yanagi, Norio haneji, Naozumi Ishimaru, Hironori Hamano and Yoshio Hayashi : Immunopathological analysis of mucosal melanocyte distribution in the human lower lip of the elderly, Pathobiology, Vol.64, No.3, 156-160, 1996.
(キーワード)
Melanocyte / Lower lip / Elderly / Sex difference
110.
Mariko Takahashi, Yasuo Mimura and Yoshio Hayashi : Role of the ICAM-1/LFA-1 pathway during development of autoimmune dacryoadenitis in an animal model for Sjogren's syndrome, Pathobiology, Vol.64, No.5, 269-274, 1996.
(要約)
We have analyzed the role of cell adhesion molecules during the development of autoimmune dacryodenitis in an NFS/sld mouse model for primary Sjögren's syndrome. The expression of cell adhesion molecules was assessed by RT-PCR and immunohistochemistry. We detected an up-regulation of local cell adhesion molecule genes (ICAM-1, LFA-1, CD44 and Mel-14) in the course of autoimmune lacrimal gland diseases. Immunohistochemically, ICAM-1 was localized exclusively in the endothelial cells of variously sized blood vessels before the onset of disease, and LFA-1, CD44 and Mel-14, expressing infiltrating cells, were found within these lesions. When the therapeutic effects of blocking cell adhesion molecules in vivo were examined, antibodies to ICAM-1 in combination with anti-LFA-1 prevented the development of autoimmune lacrimal gland diseases in NFS/sld mice. These data suggest that in Sjögren's syndrome-like autoimmune dacryoadenitis in NFS/sld mutant mice, the ICAM-1/LFA-1 pathway may play a crucial role in the development and subsequent progression of T-cell-mediated autoimmunity in the lacrimal glands.
Hironori Hamano, Norio Haneji, Kumiko Yanagi, Naozumi Ishimaru and Yoshio Hayashi : Expression of HLA-DR and cytokine genes on interferon-γ-stimulated human salivary gland cell line, Pathobiology, Vol.64, No.5, 255-261, 1996.
(要約)
Stimulation of a cultured human salivary gland (HSG) cell line by interferon (IFN)-gamma leads to HLA-DR gene expression concomitant with inflammatory cytokine genes such as IL-1 beta, tumor necrosis factor (TNF)-alpha, and IL-6 in vitro. IFN-gamma-induced HLA-DR mRNA expression was clearly detected at 2 h after the stimulation, and thereafter its level of gene expression increased until day 7 on HSG cells by reverse transcriptase (RT)-PCR. Immunofluorescence analysis revealed that cytoplasmic HLA-DR immunoreactivity was detected for the first time at 2 days after the stimulation, and its immunoreactivity increased gradually until day 7, while no immunoreactivity with HLA-DP and HLA-DQ was observed at any of the days. In addition, the expression of IL-1 beta, TNF-alpha, and IL-6 on the IFN-gamma-stimulated HSG cells was detected by immunohistochemistry and RT-PCR analysis. These results indicate that human salivary gland cells can be induced to express HLA-DR mRNA by IFN-gamma concomitant with inflammatory cytokine gene expressions such as IL-1 beta, TNF-alpha, and IL-6.
Hiroyuki Higashiyama, Ichiro Saito, Yoshio Hayashi and Nobuyuki Miyasaka : : In situ hybridization study of vascular cell adhesion molecule-1 messenger RNA expression in eheumatoid synovium, Journal of Autoimmunity, Vol.8, No.6, 947-957, 1995.
(要約)
Upregulation of vascular cell adhesion molecule-1 (VCAM-1) has been implicated in various pathological conditions. In this study, we used in situ hybridization to determine the cell types expressing VCAM-1 mRNA in rheumatoid synovium. Synovial tissues from seven rheumatoid arthritis (RA) and four osteoarthritis (OA) patients were examined for VCAM-1 mRNA expression. In situ hybridization as well as immunohistochemical analysis showed that VCAM-1 expression was mainly localized to the hyperplastic synovial lining cells and to a lesser extent endothelial cells and synovial fibroblasts in rheumatoid synovium. VCAM-1 expression in OA is less prominent. Hyperplastic synovial lining cells expressing VCAM-1 were mainly CD13+, CD14+, CD33+ and HLA-DR+. Together with morphological features, this suggests that they are type A macrophage-like synovial cells. Our findings indicate that overexpression of VCAM-1 may contribute to the formation of hyperplastic lining layers in the synovium seen in this disorder.
Yoshio Hayashi, N Haneji, K Yanagi, H Higashiyama, H Yagita and H Hamano : Prevention of adoptive transfer of murine Sjogren's syndrome into severe combined immunodeficient (SCID) mice by antibodies against intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1), Clinical and Experimental Immunology, Vol.102, No.2, 360-367, 1995.
(要約)
We have analysed the role of ICAM-1 and LFA-1 during development of autoimmune sialadenitis in MRL/lpr mice by direct analysis of RNA obtained from the salivary gland tissues, and the therapeutic effects with antibody administration on adoptive transfer system into SCID mice. The expression of cell adhesion molecules was assessed by using reverse transcriptase polymerase chain reaction (RT-PCR) and Southern blot analysis, and immunohistochemical analysis. Up-regulated expression of ICAM-1 mRNA was observed before the onset of inflammatory lesions in the salivary glands at 1 month and 2 months old, and thereafter LFA-1 mRNA was expressed within the typical inflammatory lesions, resembling human Sjögren's syndrome in MRL/lpr mice. Immunohistochemically, ICAM-1 was localized exclusively in the endothelial cells of varying sized blood vessels before the onset of disease, and LFA-1 expressing inflammatory cells were found within these lesions. When the therapeutic effects in vivo were examined, antibodies to ICAM-1 in combination with anti-LFA-1 prevented adoptive transfer of Sjögren's syndrome in MRL/lpr mice into SCID mice, while no significant effect was found when treated with either antibody. These findings indicate that in Sjögren's syndrome-like autoimmune lesions in MRL/lpr mice the ICAM-1/LFA-1 pathway may play a crucial role in the initiation and subsequent progression of T cell-mediated autoimmunity in the salivary and lacrimal glands of MRL/lpr mice.
Hiroyuki Inoue, Hitoshi Miki, Kazushi Oshimo, Katuhiro Tanaka, Yasumasa Monden, Akihiro Yamamoto, Susumu Kagawa, Nobuya Sano, Eiji Hayashi, Masaru Nagayama and Yoshio Hayashi : Familial hyperparathyroidism associated with jaw fibroma: case report and literature review, Clinical Endocrinology, Vol.43, No.2, 225-229, 1995.
(要約)
A 53-year-old female suffering from renal stones and hypercalcaemia was diagnosed as having primary hyperparathyroidism caused by hyperplasia of the parathyroid glands. She underwent total parathyroidectomy and implantation of parathyroid tissue. After one year, she underwent surgery for a jaw tumour. The pathological findings indicated it to be a cementifying fibroma. Jackson et al. (1990) reported the familial association of hyperparathyroidism with jaw tumours, and they suggested that this condition represents a new clinical syndrome. We believe that our case belongs to this syndrome.
Sachiko Iseki, Noriko Osumi-Yamashita, Kohei Miyazono, Peter Franzen, Hidenori Ichijo, Haruo Ohtani, Yoshio Hayashi and Kazuhiro Eto : Localization of transforming growth factor-β type I and type II receptors in mouse development, Experimental Cell Research, Vol.219, No.2, 339-347, 1995.
Yoshio Hayashi, Hironori Hamano, Norio Haneji, Naozumi Ishimaru and Kumiko Yanagi : Biased T cell receptor Vβ gene usage during specific stages of the development of autoimmune sialadenitis in the MRL/lpr mouse model of Sjogren's syndrome, Arthritis &Rheumatism, Vol.38, No.8, 1077-1084, 1995.
121.
Norio Haneji, Hironori Hamano, Kumiko Yanagi and Yoshio Hayashi : A new animal model for primary Sjogren's syndrome in NFS/sld mutant mice, The Journal of Immunology, Vol.153, No.6, 2796-2777, 1994.
122.
Chieri Kurashima, Masanori Utsuyama, Yoshio Hayashi and Katsuiku Hirokawa : Epithelial class II MHC antigen normally expressed by submandibular glands of elderly people, Pathobiology, Vol.62, No.5-6, 292-297, 1994.
(キーワード)
Submandibular gland / HLA-DR / HLA-DQ / HLA-DP / Class II MHC / 加齢変化 (aging patterrn) / Elderly people
123.
Maki Moritani, Katsuhiko Yoshimoto, Fumi Tashiro, Chikara Hashimoto, Jun-ichi Miyazaki, Setsuko Ii, Eiji Kudo, Hiroyuki Iwahana, Yoshio Hayashi, Toshiaki Sano and Mitsuo Itakura : Transgenic Expression of IL-10 in Pancreatic Islet A cells Accelerates Autoimmune Insulitis and Diabetes in Non-Obese Diabetic Mice., International Immunology, Vol.6, No.12, 1927-1936, 1994.
124.
Yoshio Hayashi, Norio Haneji, Hironori Hamano and Kumiko Yanagi : Transfer of Sjogren's syndrome-like autpoimmune lesions into SCID mice and prevention of lesions by anti-CD4 and anti-T cell receptor antibody treatment, Eur. J. Immunol, Vol.24, 2826-2831, 1994.
(キーワード)
Sjogren's syndrome / MRL/lpr mice / Transfer / SCID mice
Hironori Hamano, Ichiro Saito, Norio Haneji, Yuriko Mitsuhashi, Nobuyuki Miyasaka and Yoshio Hayashi : Expression of cytokine genes during development of autoimmune sialadenitis in MRL/lpr mice, European Journal of Immunology, Vol.23, No.10, 2387-2391, 1993.
Hiroyo Deguchi, Hironori Hamano and Yoshio Hayashi : c-myc, ras p21 and p53 expression in pleomorphic adenoma and its malignant form of the human salivary glands, Acta Pathologica Japonica, Vol.43, No.7-8, 413-422, 1993.
(キーワード)
c-myc / ras p21,p53 / pleomorphic adenoma
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 8396843
Hiroyo Deguchi, Hironori Hamano, Norio Haneji, Mariko Takahashi and Yoshio Hayashi : Immunopathology of phenotypic change on human parotid gland adenocarcinoma, Pathobiology, Vol.61, No.2, 83-88, 1993.
(要約)
Immunopathological analysis was made of phenotypic change in a recurrent parotid gland adenocarcinoma occurring in a patient with a long clinical course of 30 years or more. At the first and second operations, in 1959 and 1978, the resected parotid gland tumors were diagnosed histopathologically as acinic cell carcinoma. However, 11 years after the second operation, in 1989, the resected recurrent tumor showed a microscopically phenotypic change towards adenocarcinoma with typical tubular arrangement. At the last operation in 1991, histopathological examination of the tumor revealed adenocarcinoma with diffuse oncocytic change in association with cervical lymph node metastasis. These findings suggest that phenotypic change may occur in vivo among human neoplasms during a long period, which may be related to the cytodifferentiation in the salivary gland tumor.
(キーワード)
Parotid / Adenocarcinoma / Phenotype / Oncocyte
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 7692875
Youji Miyamoto, Masaru Nagayama and Yoshio Hayashi : A Cleft Child With Lobulated Tongue and Lingual Hamartoma: Report of a Case, Journal of Oral and Maxillofacial Surgery, Vol.49, No.6, 644-646, 1991.
Katsuhiro Yasuda, Kazuhito Satomura, Masaru Nagayama, Yoshio Hayashi and Atsuhito Nakanishi : The behavior of human ameloblastoma tissue in collagen matrix in vitro:an immunohistochemical study, Journal of Oral Pathology & Medicine, Vol.20, No.4, 187-190, 1991.
Yoshio Hayashi, Tamiko Takemura, Takumi Akashi, Yukiyoshi Esaki, Chieri Kurashima and Katsuiku Hirokawa : Immunopathological analysis of interstitial renal lesions in elderly people, Pathobiology, Vol.58, No.4, 230-235, 1990.
(キーワード)
Kidney / Elderly / Autoimmunity
139.
Yoshio Hayashi, Hiroyo Deguchi, Akiyo Nakahata, Chieri Kurashima and Katsuiku Hirokawa : Immunopathological study of neuropeptide expression in human salivary gland neoplasms, Pathobiology, Vol.58, No.4, 212-220, 1990.
(要約)
The immunoreactivity of anti-neuron-specific enolase (NSE) and anti-Leu-7 on formalin-fixed sections of human salivary gland neoplasms was determined by the avidin-biotin-peroxidase complex method. In addition, neuropeptides, such as vasoactive intestinal polypeptide, somatostatin, and substance P, in human salivary gland neoplasms were expressed, whereas other polypeptides, including glucagon, cholecystokinin, leu-enkephalin and calcitonin, were absent. When 182 paraffin-embedded examples of human salivary gland tumors, including 112 benign and 70 malignant neoplasms, were examined immunohistochemically, positive immunoreactivity was observed in: 51 cases with NSE (59%) and 46 cases with Leu-7 (54%) of 86 pleomorphic adenomas; 11 cases with Leu-7 (61%) of 18 Warthin's tumors; 7 cases with Leu-7 (58%) of 12 acinic cell carcinomas; 5 cases with NSE (31%) of 16 adenoid cystic carcinomas; 5 cases with NSE (42%) and 4 cases with Leu-7 (33%) of 12 adenocarcinomas; 4 cases with NSE (25%) and 6 cases with Leu-7 (38%) of 16 undifferentiated carcinomas. The other tumors, such as oxyphilic adenomas, basal cell adenomas, epidermoid carcinomas, and mucoepidermoid carcinomas, were nonreactive. Neuropeptides were observed in the neoplastic epithelial cells of certain tumors such as Warthin's tumors, acinic cell carcinomas, adenocarcinomas and undifferentiated carcinomas. These findings suggest the possibility that cells of neuroendocrine origin, present in certain neoplastic salivary gland epithelia may play a significant role in the histogenesis of human salivary gland neoplasms.
Yoshio Hayashi, M Utsuyama, C Kurashima and K Hirokawa : Spontaneous development of organ-specific autoimmune lesions in aged C57BL/6 mice, Clinical and Experimental Immunology, Vol.78, No.1, 120-126, 1989.
Yoshio Hayashi, T Hiyoshi, T Takemura, C Kurashima and K Hirokawa : Focal lymphocytic infiltration in the adrenal cortex of the elderly: immunohistological analysis of infiltrating lymphocytes, Clinical and Experimental Immunology, Vol.77, No.1, 1019-105, 1989.
Yoshio Hayashi, Tamiko Takemura and Katsuiku Hirokawa : Expression of neuron-specific enolase, Leu-7, and neuropeptides in human fetal salivary gland epithelium, The Journal of Histochemistry and Cytochemistry, Vol.37, No.7, 1147-1152, 1989.
(キーワード)
Neuropeptide / Human Fetus / Salivary Gland Epithelium / Immunoreactivity
143.
Yoshio Hayashi and K Hirokawa : Immunopathology of experimental autoallergic sialadenitis in C3H/He mice, Clinical and Experimental Immunology, Vol.75, No.3, 471-476, 1989.
Yoshio Hayashi, Akinori Kojima, Mami Hata and Katsuiku Hirokawa : A new mutation involving the sublingual gland in NFS/N mice, --- Partially arrested mucous cell differentiation ---, The American Journal of Pathology, Vol.132, No.2, 187-191, 1988.
145.
Yoshio Hayashi, C Kurashima, M Utsuyama and Katsuiku Hirokawa : Spontaneous development of autoimmune sialadenitis in aging BDF1 mice, The American Journal of Pathology, Vol.132, No.1, 173-179, 1988.
(要約)
This study reports that spontaneous autoimmune sialadenitis developed in aging female, rather than male, BDF1 mice. The lesions first appeared in 6-month-old female BDF1 mice and were aggravated with advancing age, especially in 24-month-old and 30-month-old senescent mice. In contrast, significant inflammatory changes did not develop in aging male BDF1 mice. The presence of antisalivary duct antibody was found in sera from mice with sialadenitis. The infiltrating cells in the lesions of submandibular salivary glands were mainly composed of T cells, especially Lyt 1+ and L3T4+ cells. Moreover, mild inflammatory lesions were observed in parotid, sublingual salivary glands, pancreas, or kidneys in some mice that developed spontaneously occurring sialadenitis.
Yoshio Hayashi, Tetsuo Yanagawa, Hideo Yoshida, Masayuki Azuma, Toshinobu Nishida, Yoshiaki Yura and Mitsunobu Sato : Expression of vasoactive intestinal polypeptide and amylase in a human parotid gland adenocarcinoma cell line in culture, Journal of the National Cancer Institute, Vol.79, No.5, 1025-1037, 1987.
147.
Yoshio Hayashi, Shinichi Nagamine, Tetsuo Yanagawa, Hideo Yoshida, Yoshiaki Yura, Masayuki Azuma and Mitsunobu Sato : Small cell undifferentiated carcinoma of the minor salivary gland containing exocrine, neuroendocrine, and squamous cells, Cancer, Vol.60, No.7, 1583-1588, 1987.
148.
Yoshio Hayashi, Hideo Yoshida, Shinichi Nagamine, Tetsuo Yanagawa, Yoshiaki Yura, Masayuki Azuma and Mitsunobu Sato : Induction of cells with acinar cell phenotype including presence of intracellular amylase, --- Treatment with 12-O-tetradecanoyl-phorbol-13-acetate in a neoplastic human salivary intercalated duct cell line grown in athymic nude mice ---, Cancer, Vol.60, No.5, 1000-1008, 1987.
149.
Yoshio Hayashi, Toshinobu Nishida, Hideo Yoshida, Tetsuo Yanagawa, Yoshiaki Yura and Mitsunobu Sato : Immunoreactive vasoactive intestinal polypeptide in acinic cell carcinoma of the parotid gland, Cancer, Vol.60, No.5, 962-968, 1987.
150.
Masayuki Azuma, Furumoto Nanayo, Kawamata Hitoshi, Hideo Yoshida, Yanagawa Tetsushi, Yura Yoshiaki, Yoshio Hayashi, Yoshihiro Takegawa and Mitsunobu Sato : The relation of ras oncogene product p21 expression to clinocopathological status criteria and clinical outcome in squamous cell head and neck cancer, The Cancer Journal, Vol.1, No.9, 375-380, 1987.
Tetsuo Yanagawa, Yoshio Hayashi, Hideo Yoshida, Yoshiaki Yura, Shinichi Nagamine, Takashi Bando and Mitsunobu Sato : An adenoid squamous carcinoma-forming cell line established from an oral keratinizing squamous cell carcinoma expressing carcinoembryonic antigen, The American Journal of Pathology, Vol.124, No.3, 496-509, 1986.
Yoshio Hayashi, Haruhiko Saito, Shiro Saito, Tetsuo Yanagawa, Hideo Yoshida, Yoshiaki Yura and Mitsunobu Sato : Immunoreactive somatostatin in Warthin's tumor, The American Journal of Pathology, Vol.123, No.2, 250-255, 1986.
155.
Yoshio Hayashi, Yoshiaki Yura, Hideo Yoshida, Tetsuo Yanagawa and Mitsunobu Sato : Development of allergic sialadenitis in mice immunized with mumps virus-infected submandibular salivary gland, The American Journal of Pathology, Vol.123, No.2, 271-279, 1986.
156.
Yoshio Hayashi, Hideo Yoshida, Furumoto Nanako, Yanagawa Tetsuo, Yura Yoshiaki, Yoshihiro Takegawa and Mitsunobu Sato : Monoclonal antibody analysis of peripheral blood lymphocyte subpopulations in squamous cell head and neck cancer, The Cancer Journal, Vol.1, No.1, 25-29, 1986.
157.
Masayuki Azuma, Yoshio Hayashi, Hideo Yoshida, Tetsuo Yanagawa, Yoshiaki Yura, Akemichi Ueno and Mitsunobu Sato : Emergence of differentiated subclones from a human salivary adenocarcinoma cell clone in culture after treatment with sodium butyrate, Cancer Research, Vol.46, No.2, 770-777, 1986.
Tetsuo Yanagawa, Yoshio Hayashi, Toshinobu Nishida, Hideo Yoshida, Yoshiaki Yura, Masayuki Azuma and Mitsunobu Sato : Immunohistochemical demonstration of carcinoembryonic antigen (CEA) on tissue sections from squamous cell head and neck cancer and plasma CEA levels of the patients, International Journal of Oral and Maxillofacial Surgery, Vol.15, No.3, 296-306, 1986.
M Urata, Toshinobu Nishida, Yoshio Hayashi, Hideo Yoshida, N Furumoto, T Yanagawa, Y Yura and Mitsunobu Sato : Immunoglobulin A circulating immune complexes in oral squamous cell cancer, The Cancer Journal, Vol.1, No.2, 68-71, 1986.
161.
Mitsunobu Sato, Hideo Yoshida, Yoshio Hayashi, Kenji Miyakami, Takashi Bando, Tetsuo Yanagawa, Yura Yura, Masayuki Azuma and Akemichi Ueno : Expression of epidermal growth factor and transforming growth factor-beta in human salivary gland adenocarcinoma cell line, Cancer Research, Vol.45, No.12 P 1, 6160-6167, 1985.
162.
Mitsunobu Sato, Yoshio Hayashi, Tetsuo Yanagawa, Hideo Yoshida, Yoshiaki Yura, Masayuki Azuma and Akemichi Ueno : Intermediate-sized filaments and specific markers in a human salivary gland adenocarcinoma cell line and its nude mouse tumors, Cancer Research, Vol.45, No.8, 3837-3790, 1985.
163.
Yoshio Hayashi, Tetsuo Yanagawa, Hideo Yoshida, Yoshiaki Yura, Toshiharu Nitta and Mitsunobu Sato : Induction of other differentiation stages in neoplastic epithelial duct and myoepithelial cells from the human salivary gland grown in athymic nude mice, Cancer, Vol.55, No.11, 2575-2583, 1985.
164.
Yoshio Hayashi, Mitsunobu Sato and Katsuiku Hirokawa : Induction of experimental allergic sialadenitis in mice, The American Journal of Pathology, Vol.118, No.3, 476-483, 1985.
Mitsunobu Sato, Hideo Yoshida, Yanagawa Tetsuo, Yura Yoshiaki, Urata Mitsuru, Masayuki Azuma, Furumoto Nanayo, Yoshio Hayashi and Yoshihiro Takegawa : Interferon Activity and Its Characterization in the Sera of Patients with Head and Neck Cancer, Cancer, Vol.54, No.1, 1239-1251, 1984.
Yoshio Hayashi, Toshinobu Nishida, Hideo Yoshida, Tetsuo Yanagawa, Yoshiaki Yura, Nanayo Furumoto and Mitsunobu Sato : Peripheral Tγ lymphocyte population in head and neck cancer, Cancer Immunology, Immunotherapy, Vol.17, No.3, 160-164, 1984.
(要約)
Peripheral T gamma lymphocytes were measured in head and neck cancer patients and controls. The percentage was significantly higher in the 59 cancer patients than in the 46 normal controls (P less than 0.001). The 12 patients with recurrent disease had elevated percentages of T gamma lymphocytes compared with the untreated group (n = 31; P less than 0.05) and the treated, disease-free group (n = 16; P less than 0.05). Moreover, the percentage of T gamma lymphocytes was significantly higher in the 31 patients with regional lymph node metastasis than in the node-negative group (n = 28; P less than 0.05). In a total of 37 patients with squamous cell carcinoma histologically graded I, II, and III, the absolute counts and percentages of T gamma lymphocytes in the grade I group (n = 13) showed significant decreases compared with those in the grade III group (P less than 0.05; n = 6). Moreover, postoperative serial determinations of the percentage of T gamma lymphocytes in the 14 treated, disease-free patients revealed a gradual decrease of T gamma lymphocytes, whereas the five patients with recurrent disease had a tendency to increases in the percentage of T gamma lymphocytes.
Yoshio Hayashi, Toshinobu Nishida, Hideo Yoshida, Tetsuo Yanagawa, Yoshiaki Yura, Toshiharu Nitta and Mitsunobu Sato : Peripheral K-lymphocyte population in head and neck cancer, Cancer, Vol.53, No.11, 2507-2514, 1984.
(要約)
Peripheral K-lymphocytes in head and neck cancer were measured by assay of plaque forming-cells. In the cancer patients, the percentage (4.37 +/- 0.87%, mean +/- SD, N = 42, P less than 0.01) and absolute counts (68 +/- 28/mm3, P less than 0.01) of K-lymphocytes were significantly lower than those in normal controls (8.04 +/- 1.41%; 175 +/- 53/mm3, N = 29). The untreated group showed decreased K-lymphocyte counts (63 +/- 22/mm3, N = 13) as compared with the treated, disease-free group (83 +/- 27/mm3, N = 13, P less than 0.05). There were significant correlations between absolute counts of K-lymphocytes and T-cells or B-cells within the untreated group (r = 0.79 in T-cells, P less than 0.01; r = 0.64 in B cells, P less than 0.01). Moreover, absolute counts and percentage of K-lymphocytes in the patients having regional lymph node metastasis (58 +/- 25/mm3; 4.14 +/- 0.77%, N = 22) were significantly lower than those in the negative node group (80 +/- 26/mm3, P less than 0.01; 4.63 +/- 0.91%, P less than 0.05; N = 20). In a total of 25 patients with squamous cell carcinoma who were grouped into grade I, II, and III according to classification of the histologic differentiation of the World Health Organization, the absolute counts and percentage of K-lymphocytes in the grade I group (93 +/- 33/mm3; 5.14 +/- 1.08%, N = 7) showed significant increases in comparison to those in the grade II group (62 +/- 19/mm3, P less than 0.02; 4.06 +/- 0.62%, P less than 0.02, N = 14). Moreover, the change of the K-lymphocyte population in the treated, disease-free eight patients revealed a gradual increase of K-lymphocytes. These results led us to suggest that the measurement of peripheral K-lymphocytes is a useful method of characterizing host defense in head and neck cancer.
Mitsunobu Sato, Yoshio Hayashi, Hideo Yoshida, Tetsuo Yanagawa and Yoshiaki Yura : A family with hereditary lack of T4+ inducer/helper T cell subsets in peripheral blood lymphocytes, The Journal of Immunology, Vol.132, No.3, 1071-1073, 1984.
Mitsunobu Sato, Hideo Yoshida, Yanagawa Tetsuo, Yura Yoshiaki, Urata Mitsuru, Atsumi Masahiro, Yoshio Hayashi and Yoshihiro Takegawa : Effects of intradermal administration of streptococcal preparation OK-432 on interferon and natural killer cell activities in patients with oral cancer, International Journal of Oral Surgery, Vol.13, No.1, 7-15, 1984.
Reza Shiari, Ichiro Kobayashi, Nariaki Toita, Norikazu Hatano, Nobuaki Kawamura, Motohiko Okano, Yoshio Hayashi, Kunihiko Kobayashi and Tadashi Ariga : Epitope mapping of anti-alpha-fodrin autoantibody in juvenile Sjögren's syndrome: difference in major epitopes between primary and secondary cases., The Journal of Rheumatology, Vol.33, No.7, 1395-1400, 2006.
(要約)
Juvenile Sjögren's syndrome (SS) is an early-onset type of SS. Autoantibody against the N-terminal 120 kDa form of a-fodrin is a specific and sensitive disease marker for both juvenile and adult SS. We investigated the initial and major determinants of a-fodrin in SS. Sera were obtained from patients with juvenile SS, 10 with primary SS and 10 with secondary SS. Epitope specificities of IgG antibodies were examined by dot-blot analyses using overlapping fusion proteins of the N-terminal part (561 amino acid residues) of a-fodrin as antigens. All sera from patients with primary SS reacted with amino acid residues 1 to 98 and 36 to 150, but not with 91 to 199. Epitope mapping using fusion proteins with subfragments, each consisting of about 50 amino acid residues, showed reactivity with amino acid residues 27-80 and 79-132, suggesting that at least 2 epitopes are contained in the first 150 amino acid residues. All 3 cases with neurological complications had additional epitope specificities. Sera from patients with secondary SS showed more diversified specificities and strongly reacted with amino acid residues 1-98 and 334-432, whereas the reactivities to 36-150, a major epitope in primary SS, were minimal. Major and initial B cell epitopes specifically reside in N-terminal amino acids 36-132 and could be used as a diagnostic tool for primary SS. The epitope subsequently expands to other regions of a-fodrin in association with the development of neurological complications or disease progression. Secondary SS has distinct epitope specificities.
Rieko Arakaki, Akiko Yamada, Yasusei Kudo, Yoshio Hayashi and Naozumi Ishimaru : Mechanism of Activation-Induced Cell Death of T Cells and Regulation of FasL Expression., Critical Reviews in Immunology, Vol.34, No.4, 301-314, May 2014.
(要約)
Activation-induced cell death (AICD) of T cells is a process for regulating the peripheral immune system. The fate of a T cell is controlled by numerous signals derived from various stimuli, such as antigens, cytokines, and chemokines. In healthy humans, overactivated or autoreactive T cells are harmful and are eliminated to maintain the immune system. AICD in T cells by Fas/FasL-mediated apoptosis is triggered by the switch from life to death through several signaling molecules. The control or distribution of Fas or FasL expression largely affects AICD of T cells. Although autoimmune diseases are considered to be induced by multiple factors, an impaired immune system with AICD by Fas/FasL-mediated apoptosis leads to the onset or development of autoimmunity. Based on published reports, this review describes the regulatory mechanisms involved in AICD of T cells by Fas/ FasL-mediated apoptosis and the associations between AICD and autoimmunity in humans and animal models.
Rieko Arakaki, Naozumi Ishimaru and Yoshio Hayashi : Immunotherapeutic targets in estrogen deficiency-dependent Sjögren's syndrome-related manifestations, Immunotherapy, Vol.2, No.3, 339-346, May 2010.
(要約)
Although a number of autoimmune diseases are known to develop in postmenopausal women, the mechanisms by which estrogen deficiency influences autoimmunity remain unclear. Previously, we found that tissue-specific apoptosis in the exocrine glands in estrogen-deficient mice may contribute to the development of autoimmune exocrinopathy. We found that RbAp48 overexpression induces p53-mediated apoptosis in the exocrine glands depending on estrogen deficiency. RbAp48-inducible transfectants result in rapid apoptosis with p53 phosphorylation (Ser9), and alpha-fodrin cleavage. Indeed, transgenic expression of the RbAp48 gene induced apoptosis in the exocrine glands, resulting in the development of autoimmune exocrinopathy resembling Sjögren's syndrome (SS). CD4(+) T-cell-mediated autoimmune lesions were aggravated with age, in association with production of autoantibodies against SS-A, SS-B and alpha-fodrin. These findings demonstrated that estrogen deficiency initiates tissue-specific apoptosis in the exocrine gland cells through RbAp48 overexpression and exerts a possible gender-based risk of autoimmune exocrinopathy in postmenopausal women. Thus, these data indicate RbAp48 to be a novel immunotherapeutic target for preventing epithelial cell apoptosis and the development of gender-based autoimmune exocrinopathy.
Masami Takei, Hidetaka Shiraiwa, Takashi Azuma, Yoshio Hayashi, Naoyuki Seki and Shigemasa Sawada : The possible etiopathogenic genes of Sjögren's syndrome, Autoimmunity Reviews, Vol.4, No.7, 479-484, Sep. 2005.
(要約)
Sjögren's syndrome is a chronic autoimmune disease characterized by focal lymphocytic infiltration of lacrimal and salivary glands, but the precise mechanism of this syndrome is unclear. To clarify the pathogenesis of Sjögren's syndrome, the related genes must be identified. In the present study, we investigate the increased expression of genes and molecules related to Sjögren's syndrome and present our findings of cDNA microarray analysis in the mouse model. Furthermore, we present the results of immunohistochemical analysis of salivary glands in the mouse model and patients with Sjögren's syndrome. This approach might open a new discussion of the existence of principal pathogenic molecules in Sjögren's syndrome.
(キーワード)
シェーグレン症候群 (Sjögren's syndrome) / MRL/lpr mice / NFS/sld mice / cDNA microarray / Human Homologue of SS Related Genes
Yoshio Hayashi, Rieko Arakaki and Naozumi Ishimaru : Crucial Role of Tissue-Specific Apoptosis on the Development of Primary Sjogren's Syndrome, Oral Science International, Vol.1, No.2, 55-64, Nov. 2004.
Yoshio Hayashi, Rieko Arakaki and Naozumi Ishimaru : Apoptosis and Estrogen Deficiency in Primary Sjögren Syndrome, Current Opinion in Rheumatology, Vol.16, No.5, 522-526, Sep. 2004.
(要約)
Primary Sjögren syndrome is an autoimmune disorder characterized by lymphocytic infiltrates and destruction of the salivary and lacrimal glands, and systemic production of autoantibodies to the ribonucleoprotein particles SS-A/Ro and SS-B/La. The purpose of this review is to discuss recent advances in the pathogenesis of primary Sjögren syndrome. Although several candidate autoantigens including alpha-fodrin have been reported in Sjögren syndrome, the pathogenic roles of the autoantigens in initiation and progression of SS are still unclear. It is possible that individual T cells activated by an appropriate self antigen can proliferate and form a restricted clone. Recent evidence suggests that the apoptotic pathway plays a central role in tolerizing T cells to tissue-specific self antigen, and may drive the autoimmune phenomenon. Cleavage of certain autoantigens during apoptosis may reveal immunocryptic epitopes that could potentially induce autoimmune response. The studies reviewed imply that Fas-mediated cytotoxicity and caspase-mediated alpha-fodrin proteolysis are involved in the progression of tissue destruction in Sjögren syndrome. Fas ligand (FasL), and its receptor Fas are essential in the homeostasis of the peripheral immune system. It can be considered that a defect in activation-induced cell death of effector T cells may result in the development of autoimmune exocrinopathy in Sjögren syndrome. Although the mechanisms by which estrogen deficiency influences autoimmune lesions remain unclear, it is possible that antiestrogenic actions might be a potent factor in the formation of pathogenic autoantigens.
Yoshio Hayashi, Rieko Arakaki and Naozumi Ishimaru : The role of caspase cascade on the development of primary Sjogren's syndrome, The Journal of Medical Investigation : JMI, Vol.50, No.1,2, 32-38, Feb. 2003.
(要約)
Primary Sjögren syndrome (SS) is an autoimmune disease characterized by diffuse lymphoid cell infiltrates in the salivary and lacrimal glands, resulting in symptoms of dry eye and dry mouth due to insufficient secretion. Previously, we have identified the 120 kDa alpha-fodrin as an important autoantigen on the development of SS in both animal model and SS patients, but the mechanism of a -fodrin cleavage leading to tissue destruction in SS remains unclear. In murine primary SS model, tissue-infiltrating CD4+ T cells purified from the salivary glands bear a large proportion of Fas ligand (FasL), and the salivary gland duct cells constitutively possess Fas. Infiltrating CD4+ T cells identified significant 51Cr release against mouse salivary gland (MSG) cells. In vitro studies demonstrated that apoptotic MSG cells result in a specific alpha-fodrin cleavage into 120 kDa, and preincubation with caspase-inhibitor peptides blocked alpha-fodrin cleavage. The treatment with caspase-inhibitors in vivo prevented the development of autoimmune lesions in the salivary and lacrimal glands. Thus, an increased activity in caspase cascade may be involved in the progression of alpha-fodrin proteolysis and tissue destruction on the development of SS.
Yoshio Hayashi : An essential role of apoptotic protease cascade in the pathogenesis of Sjogren's syndrome, Dentistry in Japan, Vol.37, 16-19, Mar. 2001.
Yoshio Hayashi, Kumiko Yanagi and Norio Haneji : Involvement of apoptotic protease cascade for tissue destruction in Sjogren's syndrome, Archivum Immunologiae et Therapiae Experimentalis, Vol.48, No.5, 399-403, 2000.
(要約)
Sjögren syndrome (SS) is an autoimmune disease characterized by diffuse lymphoid cell infiltrates in the salivary and lacrimal glands, resulting in symptoms of dry mouth and eyes due to insufficient secretion. Although it has been assumed that a combination of immunologic, genetic and environmental factors may play a key role in the development of autoimmune lesions in the salivary and lacrimal glands, little is known about the disease pathogenesis of SS in humans. We have identified the 120 kDa alpha-fodrin as an important autoantigen in the development of SS in both an animal model and SS patients, but the mechanism of alpha-fodrin cleavage leading to tissue destruction in SS remains unclear. Tissue-infiltrating CD4+ T cells purified from the salivary glands of a mouse model for SS bear a large proportion of Fas ligand and the salivary gland duct cells possess apoptotic receptor Fas. Anti-Fas antibody-induced apoptotic salivary gland cells result in specific alpha-fodrin cleavage to the 120 kDa fragment in vitro. Preincubation with a combination of calpain and caspase inhibitor peptides could be responsible for inhibition of the 120 kDa alpha-fodrin cleavage. Thus, an increase in apoptotic protease activities including calpain and caspases may be involved in the progression of alpha-fodrin proteolysis and tissue destruction in the development of SS.
Yoshio Hayashi : Cytokines, adhesion molecules, and immune deviation in autoimmune salivary, Oral Medicine & Pathology, Vol.1, No.1, 3-10, Jun. 1996.
(要約)
The immunopathology of most autoimmune diseases has been well defined, but the mechanisms responsible for the breakdown of self-tolerance and which lead to the development of autoimmune lesions are still unclear. Primary Sjogren's syndrome in humans is known to be an autoimmune disease characterized by diffuse lymphoid cell infiltration in the exocrine organs, involving in particular the salivary and lacrimal glands. The pathogenesis of this syndrome remains unclear, but the majority of infiltrating cells in the salivary glands were CD4^+ T cells both in humans and rodents. Since many cytokines and adhesion molecules are involved in the development of T cell-mediated autoimmunity, we have investigated local eytokine, and adhesion molecule gene expression using animal model for primary Sjogren's syndrome in MRL/lpr, and NFS/sld mice. Overexpression of interleukin-1(IL-1) β and tumor necrosis factor(TNF)-α, interferon(IFN)-γ, IL-6, IL-10, IL-12(p40) was detected in the salivary gland, and the up-regulation of ICAM-1, LFA-1, CD44, Mel-14 mRNA was found in accordance with autoimmune sialadenitis. In addition, we have analysed the expression of T cell antigen receptor(TCR)Vβ transcripts in the salivary gland tissues. Transcript for Vβ 8 was predominantly detected in the T cell infiltrating sialadenitis from the onset of the disease, suggesting that CD4^+ T cells bearing TCR Vβ 8 are play essential roles to recognize unknown autopeptide in the autoimmune salivary gland disease.
(キーワード)
シェーグレン症候群 (Sjögren's syndrome) / eytokine / adhesion molecule / T cell receptor / Animal model
Akiko Yamada, Rieko Arakaki, Mie Kurosawa, Tomoyuki Kondo, Koichi Yamada, Yoshio Hayashi and Naozumi Ishimaru : Deficient differentiation of Treg cell in a murine model of Sjogren's syndrome., 15th International Congress of Immunology, Milan, Aug. 2013.
2.
Yosuke Shikama, Naozumi Ishimaru, Yasusei Kudo, Yukiko Bandou, Nanako Aki, Yoshio Hayashi and Makoto Funaki : High Levels of Free Fatty Acids May Advance the Severity of Primary Sjögrens Syndrome, Diabetes, Vol.62, No.Supplement 1, A166, Jul. 2013.
3.
Masahiro Hiasa, Masahiro Abe, Rieko Arakaki, Akiko Yamada, Kenzo Asaoka, Toshio Matsumoto, Yoshio Hayashi and Naozumi Ishimaru : RelB attenuates the activation of the classical NF-κB pathway to facilitate osteoblastogenesis, ANZBMS Annual Scientific Meeting, Perth, Sep. 2012.
4.
Ritsuko 徳永 律子 Oura, Akiko Yamada, Rieko Arakaki, Naozumi Ishimaru and Yoshio Hayashi : Pathological Analysis of Autoimmunity in NF-B1KO/LPR mice., 8th International Congress on Autoimmunity, Granada, May 2012.
5.
Akiko Yamada, Naozumi Ishimaru, Rieko Arakaki and Yoshio Hayashi : Cathepsin L inhibition prevents murine autoimmune diabetes via suppression of CD8+T cell activity., 8th International Congress on Autoimmunity, Granada, May 2012.
6.
Kazuma Matsumoto, Naozumi Ishimaru, Takashi Izawa, Masahiro Hiasa, Yoshio Hayashi and Eiji Tanaka : Hyperfunctions of osteoclasts in a rheumatoid arthritis model, ASBMR 2011 Annual Meeting, San Diego, Sep. 2011.
7.
Kazuma Matsumoto, Naozumi Ishimaru, Yoshio Hayashi and Eiji Tanaka : Analysis of osteoclast activation in the pathogenesis of rheumatoid arthritis, International Joint Symposium: The University of Tokushima, Universitas Gadjah Mada, Niigata University, Denpasar, Bali, Dec. 2010.
8.
Ashrin Nur Meinar, Naozumi Ishimaru, Megumi Watanabe, Yoshio Hayashi and Tetsuo Ichikawa : Analysis of Molecular Mechanism for Pathogenesis of Metal Allergy, International Joint Symposium, The University of Tokushima-Niigata University-Gajah Mada University, Bali, Dec. 2010.
9.
Naozumi Ishimaru, Akiko Yamada, Rieko Arakaki, Yousuke Takahama and Yoshio Hayashi : Critical Signaling Pathway via CCR7 of Foxp3+CD25+CD4+ Regulatory T Cells for the Egress from Lymph Nodes, international immunology, Aug. 2010.
10.
Kazuma Matsumoto, Naozumi Ishimaru, Takashi Izawa, Rieko Arakaki, Ritsuko 徳永 律子 Oura, Eiji Tanaka and Yoshio Hayashi : Fas-independent apoptosis of T cells via DCs controls autoimmune arthritis, international immunology, Vol.22, No.1, 113, Aug. 2010.
11.
Ritsuko 徳永 律子 Oura, Naozumi Ishimaru, Rieko Arakaki, Takashi Izawa, Akiko Yamada, Kazuma Matsumoto, Eiji Tanaka and Yoshio Hayashi : Rapid T cell death via interaction with CD11b+ macrophages in Fas-deficient host, international immunology, Vol.22, No.1, 116, Aug. 2010.
12.
Naozumi Ishimaru, Akiko Yamada, Rieko Arakaki, Yousuke Takahama and Yoshio Hayashi : Critical signaling pathway via CCR7 of Foxp3+CD25+CD4+ regulatory T cells for the egress from lymph nodes., 第14回国際免疫学会, Kobe, Aug. 2010.
13.
Ritsuko 徳永 律子 Oura, Naozumi Ishimaru, Eiji Tanaka and Yoshio Hayashi : Rapid peripheral T cell death in Fas-deficient mice, 88th IADR, Barcelona, Jul. 2010.
Emi Kawakami, Nao Kinouchi, Adachi Taro, Yutaka Ohsawa, Naozumi Ishimaru, Hideyo Ohuchi, Yoshihide Sunada, Yoshio Hayashi, Eiji Tanaka and Sumihare Noji : Special Processed Collagen-mediated Application of Myostatin-siRNA for Muscular Atrophy Diseases, 88th IADR, Barcelona, Jul. 2010.
16.
Emi Kawakami, Nao Kinouchi, Yutaka Osawa, Naozumi Ishimaru, Hideyo Ohuchi, Yoshihide Sunada, Yoshio Hayashi, Eiji Tanaka and Sumihare Noji : Strategic study of atelocollagen-mediated application of mystatin-targeting siRNA for therapeutic use for muscular atrophy diseases, QOL International Congress, Niigata, Feb. 2010.
17.
Takashi Sakai, HoangNam Tran, Sun Mi Kim, Li Liu, Xichuan Teng, Yuji Shishido, Mukai-Sakai Rika, Mitsuru Matsumoto, Kazunori Ishimura, Yoshio Hayashi, Ryuji Kaji and Kiyoshi Fukui : Nucling, a novel stress-sensitive protein, regulates NF-kappa B activation, The 4th International Congress on Stress Responses in Biology and Medicine, The 4th Annual Meeting of the Biomedical Society for Stress Response, Sapporo, Oct. 2009.
18.
Takeo Iwata, Masamichi Kuwajima, Akiko Sukeno, Naozumi Ishimaru, Yoshio Hayashi, M Wabitsch, Noriko Mizusawa, Mitsuo Itakura and Katsuhiko Yoshimoto : YKL-40 secreted from macrophages infiltrating into adipose tissue inhibits degradation of type I collagen., International symposium on diabetes, Tokyo, Mar. 2009.
19.
Takashi Sakai, Li Liu, Xichuan Teng, Naozumi Ishimaru, Rika Sakai, HoangNam Tran, Nobuya Sano, Yoshio Hayashi, Ryuji Kaji and Kiyoshi Fukui : Inflammatory disease and cancer with a decrease in Kupffer cell numbers in Nucling-konckout mouse, 20th FAOBMB Taipei Conference, Taipei, Oct. 2008.
Yoshio Hayashi and Naozumi Ishimaru : Development of autoimmune expcrinopathy resembling Sjogren's syndrome in RbAp48 transgenic mice., Boston, Nov. 2007.
22.
Takashi Izawa, Naozumi Ishimaru, Keiji Moriyama and Yoshio Hayashi : Immunoregulatory Role of RANKL-stimulated Dendritic Cells on Autoimmune Arthritis in MRL/lpr Mice, ACR/ARHP 71th Annual Scientific Meeting, Vol.56, No.Suppl 9, S-105, Boston, Nov. 2007.
23.
Yoshio Hayashi and Naozumi Ishimaru : Developmant of autoimmune excrinopathy resembling Sjorgen`s syndrome in RbAp48 transgenic mice., The 71st Annual meeting of the American College of Rheumatology., Bostone,USA., Nov. 2007.
24.
Takashi Izawa, Naozumi Ishimaru, Keiji Moriyama and Yoshio Hayashi : Immunoregulatory role of RANKL-stimulated dendritic cells on autoimmune arthritis in MRL/lpr mice, 29th Annual Meeting of the American Society for Bone and Mineral Research, Vol.22, No.SUPPL 1, S270, Honolulu, Sep. 2007.
25.
Jun Ooshima, Takashi Izawa, Naozumi Ishimaru, Yoshio Hayashi and Keiji Moriyama : Immunoregulatory Effects of Dendritic Cells Stimulated with RANKL on the Development of Autoimmune Disease in Arthritis Model Mice, The 1st International Symposium and Workshop "The Future Direction of Oral Sciences in the 21st Century", 55, Awaji, Mar. 2007.
26.
Yoshio Hayashi, N Ishimaru and N Katunuma : Cathepsin L-inhibitor prevents proinsulin peptide processing and autimmune diabetes in nonobase diabetuc(NOD)mice., International Sympojium on Medical and biological Perspectives in proteases and Their Inhibitors., Awaji ,Japan, Jun. 2006.
27.
Yoshio Hayashi : NFS/sld mouse model for primary Sjogren's syndrome., 8th International Symposium on Sjogren's Syndrome, Washington, D.C., Apr. 2006.
28.
Yoshio Hayashi and N Ishimaru : Involvement of interstitial oneumoina in Sjogren`s syndrome mouse model with aging., --- T Cell Immunity & Aging. ---, Freudenstadt-Lauterbad,Germany, Dec. 2005.
29.
Yoshio Hayashi and Naozumi Ishimaru : Involvement of Interstitial Pneumonia in Sjogren's Syndrome Mouse Model with Aging., International Symposium on T Cells in Aging., Stuttgart, Dec. 2005.
30.
Yoshio Hayashi, R Arakaki and N Ishimaru : Possible mechanism of autoimmune exocrinopathy resembling Sjogren`s syndrome through RbAp48 overexpression., --- Gordon Research Conference on Salivary Glands & Exocrine Secretion. ---, Ventura,USA., Feb. 2005.
31.
Yoshio Hayashi, Rieko Arakaki and Naozumi Ishimaru : Possible Mechanism of Autoimmune Exocrinopathy Resembling Sjogren's Syndrome through RbAp48 Overexpression., Gordon Research Conference., Ventura, Feb. 2005.
32.
Yoshio Hayashi and N Ishimaru : Development of autoimmune arthritis in Sjogren`s syndrome mouse model with aging via bystander T cell activation, 12th International Congress of Immunology, Montreal, Jul. 2004.
33.
Takashi Izawa, Keiji Moriyama and Yoshio Hayashi : Role of Dendritic Cells Stimulated with RANKL for Modulation of Autoimmune Arthritis in MRL/lpr Mice, 25th Annual Meeting of the American Society for Bone and Mineral Research, Vol.18, No.SUPPL 2, S396, Minneapolis, Sep. 2003.
34.
Yoshio Hayashi, N Ishimaru, R Arakaki and N Katunuma : Critical role of cathepsin inhibitors on autoantigen processing and autoimmunity, 44th International Symposium on Regulation of Enzyme Activity and Synthesis, Indianapolis,USA, Sep. 2003.
35.
Yoshio Hayashi and N Ishimaru : Role of estrogen-deficiency on the development of salivary gland autoimmunity, Gordon Research Conference, Ventura,USA, Feb. 2003.
36.
Yoshio Hayashi and N Ishimaru : Induction of oral tolerance with epitope peptide of autoantigen in the aged mouse model for primary Sjogren`s syndrome, 3rd European Congress of Gerontology, Frenze,Italy, Nov. 2002.
37.
Takashi Izawa, Keiji Moriyama and Yoshio Hayashi : Acceleration of Autoimmune Arthritis in MRL/lpr Mice Transferred with RANKL-stimulated Dendritic cells, 24th Annual Meeting of the American Society for Bone and Mineral Research, Vol.17, No.SUPPL 1, S490, San Antonio, Sep. 2002.
38.
Yoshio Hayashi : Pathogenic T cell epitope on alpha-fodrin autoantigen in Sjogren`s syndrome, VIIIth International Symposium on Sjogren`s Syndrome, Kanazawa, May 2002.
39.
Yoshio Hayashi : Immunodominant T cell epitope on alpha-fodrin autoantigen in primary Sjogren`s syndrome, 1st International Congress on Salivary Gland Diseases, Genwva,Switzerland, Jan. 2002.
40.
Yoshio Hayashi : Acceleration of autoimmune exocrinopathy with aging in murine Sjogren`s syndrome through Fas-medeated apoptosis, The 17th Congress of the International Association of Gerontology, Vancouver, Jul. 2001.
41.
K Yanagi, N Haneji, I Saito and Yoshio Hayashi : Autoreactive T cell clone against alpha-fodrin in Sjogren`s syndrome, XXIII International Congress of the International Academy of Pathology, Nagoya, Oct. 2000.
42.
Yoshio Hayashi : Breakdown product of alpha-fodrin as an important autoantigen in Sjogren's syndrome, VIIth International Symposium on Sjogren's Syndrome, Venice,Italy, Dec. 1999.
43.
Yoshio Hayashi : The role for alpha-fodrin as an autoantigen in Sjogren's syndrome, Gordon Research Conference, Ventura,USA, Feb. 1999.
44.
Yoshio Hayashi : Alpha-fodrin as a potential candidate autoantigen in Sjogren's sndrome, VIth International Symposium on Sjogren's Syndrome, Avon,USA, Oct. 1997.
45.
Yoshio Hayashi : A new mouse model of primary Sjogren`s syndrome, International Conference on Immunopathology of Mucous Membrane and Exocrine Organs, Bergen,Norway, May 1997.
46.
Yoshio Hayashi, N Haneji and N Ishimaru : Acceleration of age-related autoimmunity in MRL/+mice through oophorectomy, International Conference on Immunology and Aging, Bethesda,Maryland,USA, Jun. 1996.
47.
Maki Moritani, Katsuhiko Yoshimoto, J. Miyazaki, F. Tashiro, Setsuko Ii, Eiji Kudo, Hiroyuki Iwahana, Yoshio Hayashi, Toshiaki Sano and Mitsuo Itakura : (Workshop) Local Production of IL-10 in Pancreatic Islet A-Cells in Transgenic NOD Mice Accelerates Autoimmune Insulitis and Diabetes., Combined Meeting of the 8th International Lymphokine Workshop and The 4th International Workshop on Cytokines, Osaka, Oct. 1993.
新垣 理恵子, 江口 洋, 山田 安希子, 工藤 保誠, 三田村 佳典, 林 良夫, 石丸 直澄 : Eye drop administration of rebamipide is effective to a dry eye symptom in a model mice of Sjogren's syndorome., 日本免疫学会総会·学術集会記録, 2013年12月.
Rieko Arakaki, Hiroshi Eguchi, Akiko Yamada, Yasusei Kudo, Yoshinori Mitamura, Yoshio Hayashi and Naozumi Ishimaru : An Effective Therapy of Ocular Lesions in a Murine Model for Sjogren's Syndorome by Eye Drop Administration of Rebamipide., Proceedings of the Japanese Society for Immunology, Dec. 2012.
11.
Akiko Yamada, Rieko Arakaki, Mie Kurosawa, Tomoyuki Kondo, Yasusei Kudo, Yoshio Hayashi and Naozumi Ishimaru : Functional defect of Foxp3+Treg cells in Sjogren's syndrome., Proceedings of the Japanese Society for Immunology, Dec. 2012.
松本 一真, 新垣 理恵子, 山田 安希子, 日浅 雅博, 大浦 徳永 律子 律子, 岩浅 亮彦, 田中 栄二, 林 良夫, 石丸 直澄 : Hyperfunctions of osteoclasts in pathogenesis of rheumatoid arthritis in MRL/lpr mice., 第40回日本免疫学会学術集会, 2011年11月.
18.
日浅 雅博, 新垣 理恵子, 山田 安希子, 大浦 徳永 律子 律子, 安倍 正博, 松本 俊夫, 林 良夫, 石丸 直澄, 淺岡 憲三 : A novel role of NF-B relB in bone remodeling., 第40回日本免疫学会学術集会, 2011年11月.
19.
Ritsuko 徳永 律子 Oura, Rieko Arakaki, Masahiro Hiasa, Akiko Yamada, Eiji Tanaka, Yoshio Hayashi and Naozumi Ishimaru : In vivo T cell apoptosis via interaction with CD11b+ macrophages in Fas-deficient host., Proceedings of the Japanese Society for Immunology, Vol.40, 111, Nov. 2011.
20.
Akihiko Iwasa, Rieko Arakaki, Akiko Yamada, Eiji Tanaka, Yoshio Hayashi and Naozumi Ishimaru : A critical role of aromatase in the pathogenesis of Sjogren's syndrome., Proceedings of the Japanese Society for Immunology, Vol.40, 127, Nov. 2011.
21.
Rieko Arakaki, Akiko Yamada, Yoshio Hayashi and Naozumi Ishimaru : The role of Th17 cell on the development of Sjogren's syndrome in estrogen deficient NOD mice., Proceedings of the Japanese Society for Immunology, Vol.40, 128, Nov. 2011.
22.
Nur Meinar Ashrin, Megumi Watanabe, Rieko Arakaki, Akiko Yamada, Tetsuo Ichikawa, Yoshio Hayashi and Naozumi Ishimaru : Analysis of Immune Responess to Metal using Murine Nickel Allergy Model., Proceedings of the Japanese Society for Immunology, Vol.40, 102, Nov. 2011.
Akiko Yamada, Naozumi Ishimaru, Rieko Arakaki and Yoshio Hayashi : Therapeutic Analysis by Selective Inhibition of Cathepsin L for Autoimmune Diabetes in NOD Mice, 日本免疫学会総会・学術集会, Dec. 2009.
43.
Kazuma Matsumoto, Naozumi Ishimaru, Takashi Izawa, Rieko Arakaki, Ritsuko 徳永 律子 Oura, Eiji Tanaka and Yoshio Hayashi : Immunoregulatory effect via RANKL and Fas signaling of dendritic cells in immurine autoimmune arthritis models, 日本免疫学会総会・学術集会, Dec. 2009.
44.
Ritsuko 徳永 律子 Oura, Naozumi Ishimaru, Akiko Yamada, Rieko Arakaki, Takashi Izawa, Kazuma Matsumoto, Eiji Tanaka and Yoshio Hayashi : Immune Regulation by Macrophages through cross-talk between Fas and NF-B Signaling, 日本免疫学会総会・学術集会, Dec. 2009.
45.
Naozumi Ishimaru, Rieko Arakaki, Akiko Yamada, Yousuke Takahama and Yoshio Hayashi : In situ patrolling of regulatory T cells is essential for protecting organ-specific autoimmunity, 日本免疫学会総会・学術集会, Dec. 2009.
46.
Masayuki Kohashi, Naozumi Ishimaru, Akiko Yamada, Rieko Arakaki and Yoshio Hayashi : Neonatal exposure to low-dose 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD)causes autoimmunity due to the disruption of Tcell tolerance, 2009日本免疫学会総会・学術集会記録, Vol.39, Dec. 2009.
井澤 俊, 石丸 直澄, 新垣 理恵子, 大浦 徳永 律子 律子, 田中 栄二, 林 良夫 : Analysis of RANKL and Fas signaling through NF-kB in dendritic cell activation in murine autoimmune arthritis models, 日本免疫学会総会・学術集会記録, Vol.38, 176, 2008年12月.
54.
Rieko Arakaki, Naozumi Ishimaru, Akiko Yamada and Yoshio Hayashi : Identification and functional analysis of CD11c+B220+ dendritic cells induced by estrogen deficiency in salivary glands ., 第38回日本免疫学会総会・学術集会, Vol.38, 97, Dec. 2008.
55.
Naozumi Ishimaru, Kohashi Masayuki, Akiko Yamada, Rieko Arakaki and Yoshio Hayashi : Neonatal exposure to low-dose 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD)causes autoimmunity due to the disruption of T cell tolerance, 第38回日本免疫学会総会・学術集会, Vol.38, 97, Dec. 2008.
56.
Akiko Yamada, Naozumi Ishimaru, Rieko Arakaki and Yoshio Hayashi : Therapeutic analysis for autoimmune diabetes in NOD mice using specific inhibitor for cathepsin L, 第38回日本免疫学会総会・学術集会, Vol.38, 93, Dec. 2008.
Naozumi Ishimaru, Rieko Arakaki and Yoshio Hayashi : IFN-γ-producing epithelial cells links autoimmune exocrinopathy resembling SS in RbAp48 TG mice, 第37回日本免疫学会総会・学術集会, Vol.37, 263, Nov. 2007.
67.
Rieko Arakaki, Naozumi Ishimaru, Akiko Yamada and Yoshio Hayashi : Role of estrogen deficiency for aberrant MHC class 2 expression in the exocring gland cells, 第37回日本免疫学会総会・学術集会, Vol.37, 265, Nov. 2007.
68.
Akiko Yamada, Naozumi Ishimaru, Rieko Arakaki and Yoshio Hayashi : T cell functions with aging in NF-kB1-deficient mice, 第37回日本免疫学会総会・学術集会, Vol.37, 174, Nov. 2007.
69.
Takashi Izawa, Naozumi Ishimaru, Masayuki Kohashi, Rieko Arakaki and Yoshio Hayashi : Crosstalk between RANKL and Fas signaling in dendritic cells controls immune tolerance, 第37回日本免疫学会総会・学術集会 ワークショップ, Vol.37, Nov. 2007.
70.
Miho Okuno, Naozumi Ishimaru, Takashi Izawa, Rieko Arakaki, Akiko Yamada and Yoshio Hayashi : Analysis of NF-kB in T cell activation in murine autoimmune model, 第37回日本免疫学会総会・学術集会, Vol.37, 119, Nov. 2007.
71.
Masayuki Kohashi, Naozumi Ishimaru, Akiko Yamada, Rieko Arakaki, Jun Kanno and Yoshio Hayashi : The effect of TCDD administration on murine model for Sjogren's syndrome, 第37回日本免疫学会総会・学術集会, Vol.37, 116, Nov. 2007.
Yoshio Hayashi, Naozumi Ishimaru and Nobuhiko Katunuma : Cathepsin L-inhibitor Prevents Proinsulin Peptide Processing and Autoimmune Diabetes in Nonobetic(NOD)Mice, Satellite Meeting of the 20th IUBMB International Congress and 11th FAOBMB Congress, Jun. 2006.
91.
Naozumi Ishimaru and Yoshio Hayashi : In Vivo Role of NF-κB-inducing Kinase for T Cell Activation, Satellite Meeting of the 20th IUBMB International Congress and 11th FAOBMB Congress, Jun. 2006.
Naozumi Ishimaru, Hidehiro Kishimoto, Jonathan Sprent and Yoshio Hayashi : Immunoregulation by NF-kB for T cell activation in autoimmune disease, 日本免役学会総会·学術集会記録, Vol.35, 275, Dec. 2005.
99.
Naozumi Ishimaru, Hidehiro Kishimoto, Jonathan Sprent and Yoshio Hayashi : Immunoregulation by NF-kB for T cell activation in autoimmune disease, 第35回日本免疫学会総会・学術集会 ワークショップ, Vol.35, 275, Dec. 2005.
Yoshio Hayashi, Toshinobu Nishida, Tetsuo Yanagawa, Hideo Yoshida and Mitsunobu Sato : Expression of carcinoemryonic antigen in the neoplastic cells originated from human salivary gland, Tissue Culture in Dentistry, Vol.19, 27-28, 1982.
153.
Yoshio Hayashi, Toshinobu Nishida, Toshiharu Nitta, Tetsuo Yanagawa, Hideo Yoshida and Mitsunobu Sato : Peripheral k lymphocytes as an immunologic parameter in the patients with oral cancer, Tissure Culture in Dentistry, Vol.19, 25-26, 1982.