248 results on '"Hachimura, S."'
Search Results
152. IgA production in the large intestine is modulated by a different mechanism than in the small intestine: Bacteroides acidifaciens promotes IgA production in the large intestine by inducing germinal center formation and increasing the number of IgA+ B cells.
- Author
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Yanagibashi T, Hosono A, Oyama A, Tsuda M, Suzuki A, Hachimura S, Takahashi Y, Momose Y, Itoh K, Hirayama K, Takahashi K, and Kaminogawa S
- Subjects
- Animals, B-Lymphocytes metabolism, B-Lymphocytes pathology, Bacteroides Infections microbiology, Germinal Center metabolism, Germinal Center pathology, Immunity, Mucosal, Immunoglobulin A biosynthesis, Intestinal Mucosa immunology, Intestinal Mucosa metabolism, Intestinal Mucosa microbiology, Intestinal Mucosa pathology, Intestine, Large metabolism, Intestine, Large microbiology, Intestine, Large pathology, Intestine, Small metabolism, Intestine, Small microbiology, Intestine, Small pathology, Lactobacillus immunology, Mice, Mice, Inbred BALB C, Organ Specificity immunology, Antibody Formation, B-Lymphocytes immunology, Bacteroides immunology, Bacteroides Infections immunology, Germinal Center immunology, Immunoglobulin A immunology, Intestine, Large immunology, Intestine, Small immunology
- Abstract
It has been demonstrated that intestinal commensal bacteria induce immunoglobulin (Ig) A production by promoting the development of gut-associated lymphoid tissues in the small intestine. However, the precise mechanism whereby these bacteria modulate IgA production in the large intestine, which harbors the majority of intestinal commensals, is poorly understood. In addition, it is not known which commensal bacteria induce IgA production in the small intestine and which induce production in the large intestine. To address these issues, we generated gnotobiotic mice mono-associated with different murine commensal bacteria by inoculating germ-free (GF) mice with Lactobacillus johnsonii or Bacteroides acidifaciens. In GF mice, IgA production was barely detectable in the small intestine and was not detected in the large intestine. Interestingly, total IgA secretion in the large intestinal mucosa of B. acidifaciens mono-associated (BA) mice was significantly greater than that of GF and L. johnsonii mono-associated (LJ) mice. However, there was no difference in total IgA production in the small intestine of GF, LJ and BA mice. In addition, in the large intestine of BA mice, the expression of IgA(+) cells and germinal center formation were more remarkable than in GF and LJ mice. Furthermore, B. acidifaciens-specific IgA was detected in the large intestine of BA mice. These results suggest that the production of IgA in the large intestine may be modulated by a different mechanism than that in the small intestine, and that B. acidifaciens is one of the predominant bacteria responsible for promoting IgA production in the large intestine., (Copyright © 2012 Elsevier GmbH. All rights reserved.)
- Published
- 2013
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153. Memory B cells in the lung participate in protective humoral immune responses to pulmonary influenza virus reinfection.
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Onodera T, Takahashi Y, Yokoi Y, Ato M, Kodama Y, Hachimura S, Kurosaki T, and Kobayashi K
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- Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Humans, Immunoglobulin A immunology, Immunoglobulin G immunology, Lung cytology, Antibody Formation, B-Lymphocytes immunology, Immunologic Memory, Influenza, Human immunology, Lung immunology, Lung Diseases immunology
- Abstract
After pulmonary virus infection, virus-binding B cells ectopically accumulate in the lung. However, their contribution to protective immunity against reinfecting viruses remains unknown. Here, we show the phenotypes and protective functions of virus-binding memory B cells that persist in the lung following pulmonary infection with influenza virus. A fraction of virus-binding B-cell population in the lung expressed surface markers for splenic mature memory B cells (CD73, CD80, and CD273) along with CD69 and CXCR3 that are up-regulated on lung effector/memory T cells. The lung B-cell population with memory phenotype persisted for more than 5 mo after infection, and on reinfection promptly differentiated into plasma cells that produced virus-neutralizing antibodies locally. This production of local IgG and IgA neutralizing antibody was correlated with reduced virus spread in adapted hosts. Our data demonstrates that infected lungs harbor a memory B-cell subset with distinctive phenotype and ability to provide protection against pulmonary virus reinfection.
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- 2012
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154. Two distinct epitopes on the ovalbumin 323-339 peptide differentiating CD4⁺T cells into the Th2 or Th1 phenotype.
- Author
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Nakajima-Adachi H, Koike E, Totsuka M, Hiraide E, Wakatsuki Y, Kiyono H, and Hachimura S
- Subjects
- Amino Acid Sequence, Animals, CD4-Positive T-Lymphocytes immunology, Epitopes chemistry, Food Hypersensitivity immunology, Mice, Molecular Sequence Data, Peptide Fragments chemistry, Phenotype, CD4-Positive T-Lymphocytes cytology, Cell Differentiation immunology, Epitopes immunology, Ovalbumin chemistry, Peptide Fragments immunology, Th1 Cells cytology, Th2 Cells cytology
- Abstract
The epitopes for OVA323-339-specific CD4⁺T cells from OVA23-3 food allergy model and DO11.10 tolerant induction model mice were analyzed. We found that OVA23-3 CD4⁺T cells recognized the N-terminal region, showing strong proliferation and the Th2-phenotype, and that DO11.10 CD4⁺T cells recognized the C-terminal region, showing milder proliferation and a Th1-skewed response. These differences may regulate the responses of those mice to OVA-feeding, inflammation and tolerance.
- Published
- 2012
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155. Naïve CD4+ T cells of Peyer's patches produce more IL-6 than those of spleen in response to antigenic stimulation.
- Author
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Hashiguchi M, Hachimura S, Ametani A, Sato T, Kojima H, Kumagai Y, Habu S, Kobata T, and Kaminogawa S
- Subjects
- Animals, Antigen-Presenting Cells immunology, Antigens immunology, Dendritic Cells immunology, GA-Binding Protein Transcription Factor metabolism, Immunity, Mucosal immunology, Interleukin-5 analysis, Interleukin-6 analysis, Mice, Mice, Transgenic, Receptors, Antigen, T-Cell immunology, Receptors, Antigen, T-Cell metabolism, Interleukin-5 biosynthesis, Interleukin-6 biosynthesis, Peyer's Patches cytology, Peyer's Patches immunology, Peyer's Patches metabolism, Spleen cytology, Spleen immunology, Spleen metabolism, T-Lymphocytes immunology, T-Lymphocytes metabolism
- Abstract
Peyer's patches (PPs) are potential sites where specific mucosal immune responses and oral tolerance are induced. The unique features of these immune responses are thought to occur in micromilieu and are largely affected by antigen-presenting cells (APCs) such as dendritic cells. In this study, we investigated the cytokine profiles induced by the activation of CD4(+) T cells of PPs. PP cells from TCR transgenic mice secreted greater amounts of IL-5 and IL-6 than spleen cells after antigenic stimulation. IL-5 was mainly produced by PP non-T cells, whereas IL-6 was secreted by PP CD4(+) cells. PPs contained two major populations including naïve and memory/activated CD4(+) cells; both populations secreted IL-6 upon activation. We also found that CD4(+)/CD62L(hi) naïve cells from PPs secreted a greater amount of IL-6 after stimulation than those from the spleen. Furthermore, subtraction and qPCR analyses revealed that PP CD4(+)/CD62L(hi) cells express a greater amount of transcripts of GA-binding protein β subunit 1 than those of the spleen. These results suggest that naïve T cells as well as non-T cells and activated/memory T cells from PPs are distinct from their splenic counterparts and thus cause unique immune responses the in intestine., (Copyright © 2011 Elsevier B.V. All rights reserved.)
- Published
- 2011
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156. Oral tolerance induction does not resolve gastrointestinal inflammation in a mouse model of food allergy.
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Burggraf M, Nakajima-Adachi H, Hachimura S, Ilchmann A, Pemberton AD, Kiyono H, Vieths S, and Toda M
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- Administration, Oral, Animals, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes metabolism, Cytokines metabolism, Disease Models, Animal, Egg White, Female, Gastroenteritis pathology, Immunoglobulin E, Interleukin-10 metabolism, Intestine, Small immunology, Intestine, Small pathology, Mice, Mice, Inbred BALB C, Ovalbumin adverse effects, Spleen immunology, Food adverse effects, Food Hypersensitivity immunology, Gastroenteritis immunology, Immune Tolerance immunology
- Abstract
Scope: Oral immunotherapy (OIT) involving continuous oral administration of allergenic foods has gained attention as a therapy for food allergies. To study the influence of oral administration of allergenic foods on gastrointestinal symptoms including inflammation, we established a mouse model of food-induced intestinal allergy., Methods and Results: BALB/c mice were fed an egg white (EW) diet containing ovalbumin (OVA, a major EW allergen) after intraperitoneal sensitisation with OVA and Alum. The mice on the EW diet for one wk presented gastrointestinal symptoms (i.e. weight loss and soft stools) and inflammation in the small intestines (i.e. duodenum, jejunum and ileum). Further continuous EW diet resolved the weight loss but not the soft stools. Splenic CD4(+) T-cells of EW diet-fed mice on the continuous diet showed less proliferation and cytokine production compared with those of control mice, suggesting tolerance induction by the diet. The continuous EW diet reduced levels of OVA-specific IgE antibodies, but significantly aggravated the inflammation in the jejunum., Conclusion: Our mouse model would be useful to investigate inflammatory and regulatory mechanisms in food-induced intestinal allergies. Our results suggest potential gastrointestinal inflammation in patients undergoing OIT as continuous administration of allergenic foods, even though the therapy may induce clinical tolerance., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2011
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157. Orally administered Bifidobacterium triggers immune responses following capture by CD11c(+) cells in Peyer's patches and cecal patches.
- Author
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Hiramatsu Y, Hosono A, Konno T, Nakanishi Y, Muto M, Suyama A, Hachimura S, Sato R, Takahashi K, and Kaminogawa S
- Abstract
We have investigated the immunomodulatory mechanisms of Bifidobacterium pseudocatenulatum JCM7041 (Bp) as model of probiotics following oral administration to mice. This study was conducted with the aim of clarifying the mechanism of immunomodulation induced by oral administration of probiotic bacteria through elucidation of the detailed mechanism of transfer of orally administered bacterial cells within the body and the interaction between bacterial cells and cells of the immune tissues. We observed the localization of Bp in mice following oral administration, showing that Bp was surrounded by CD11c(+) cells in Peyer's patches (PP) and cecal patches (CP). These results indicated that Bp might induce CD11c(+) cell-mediated immune responses directly. Furthermore, IL-10 and IL-12p40 production by Thy1.2(-) cells, including CD11c(+) cells, increased significantly. Production of IL-10 and IL-12p40 by bone marrow-derived dendritic cells (BMDC) was significantly increased by Bp stimulation. These results suggest that oral administration of Bp induces immune responses directly following capture by CD11c(+) dendritic cells (DCs). Subsequently, we observed oral administration of Bp for 1 week induced IgA and IgA-associated cytokine production by CP and PP cells, suggesting that Bp induced DC-mediated immune responses on CP as well as PP.
- Published
- 2011
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158. Intestinal commensal bacteria promote T cell hyporesponsiveness and down-regulate the serum antibody responses induced by dietary antigen.
- Author
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Tsuda M, Hosono A, Yanagibashi T, Kihara-Fujioka M, Hachimura S, Itoh K, Hirayama K, Takahashi K, and Kaminogawa S
- Subjects
- Animals, Antigens immunology, Food Hypersensitivity etiology, Food Hypersensitivity immunology, Germ-Free Life, Humans, Intestines immunology, Mice, Mice, Inbred BALB C, Mice, Transgenic, Ovalbumin immunology, Receptors, Antigen, T-Cell genetics, Antibodies blood, Dietary Proteins immunology, Down-Regulation, Intestines microbiology, T-Lymphocytes immunology
- Abstract
Colonization of the gut by commensal bacteria modulates the induction of oral tolerance and allergy. However, how these intestinal bacteria modulate antigen-specific T cell responses induced by oral antigens remains unclear. In order to investigate this, we used germ-free (GF) ovalbumin (OVA)-specific T cell receptor transgenic (OVA23-3) mice. Conventional (CV) or GF mice were administered an OVA-containing diet. Cytokine production by CD4(+) cells from spleen (SP), mesenteric lymph nodes (MLN) and Peyer's patches (PP) was evaluated by ELISA, as was the peripheral antibody titer. T cell phenotype was assessed by flow cytometry. CD4(+) cells from the SP and MLN of CV and GF mice fed an OVA diet for 3 weeks produced significantly less IL-2 than the corresponding cells from mice receiving a control diet, suggesting that oral tolerance could be induced at the T cell level in the systemic and intestinal immune systems of both bacterial condition of mice. However, we also observed that the T cell hyporesponsiveness induced by dietary antigen was delayed in the systemic immune tissues and was weaker in the intestinal immune tissues of the GF mice. Intestinal MLN and PP CD4(+) T cells from these animals also produced lower levels of IL-10, had less activated/memory type CD45RB(low) cells, and expressed lower levels of CTLA-4 but not Foxp3 compared to their CV counterparts. Furthermore, GF mice produced higher serum levels of OVA-specific antibodies than CV animals. CD40L expression by SP CD4(+) cells from GF mice fed OVA was higher than that of CV mice. These results suggest that intestinal commensal bacteria promote T cell hyporesponsiveness and down-regulate serum antibody responses induced by dietary antigens through modulation of the intestinal and systemic T cell phenotype., (Copyright 2010 Elsevier B.V. All rights reserved.)
- Published
- 2010
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159. IL-10 and IL-27 producing dendritic cells capable of enhancing IL-10 production of T cells are induced in oral tolerance.
- Author
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Shiokawa A, Tanabe K, Tsuji NM, Sato R, and Hachimura S
- Subjects
- Administration, Oral, Animals, DNA-Binding Proteins genetics, DNA-Binding Proteins immunology, Dendritic Cells metabolism, Forkhead Transcription Factors immunology, Forkhead Transcription Factors metabolism, Immune Tolerance immunology, Interleukin-10 immunology, Interleukins immunology, Lymph Nodes immunology, Lymph Nodes metabolism, Mice, Mice, Inbred BALB C, Mice, Knockout, Mice, Transgenic, Ovalbumin immunology, Peyer's Patches immunology, Peyer's Patches metabolism, Receptors, Antigen, T-Cell immunology, Receptors, Antigen, T-Cell metabolism, Spleen immunology, Spleen metabolism, T-Lymphocytes metabolism, DNA-Binding Proteins metabolism, Dendritic Cells immunology, Interleukin-10 metabolism, Interleukins metabolism, Mouth immunology, T-Lymphocytes immunology
- Abstract
Oral tolerance is a key feature of intestinal immunity, generating systemic tolerance to ingested antigens (Ag). Dendritic cells (DC) have been revealed as important immune regulators, however, the precise role of DC in oral tolerance induction remains unclear. We investigated the characteristics of DC in spleen, mesenteric lymph node (MLN), and Peyer's patch (PP) after oral Ag administration in a TCR-transgenic mouse model. DC from PP and MLN of tolerized mice induced IL-10 production but not Foxp3 expression in cocultured T cells. IL-10 production was markedly increased after 5-7-day Ag administration especially in PP DC. On the other hand, IL-27 production was increased after 2-5-day Ag administration. CD11b(+) DC, which increased after ingestion of Ag, prominently expressed IL-10 and IL-27 compared with CD11b(-) DC. These results suggest that IL-10 and IL-27 producing DC are increased by interaction with antigen specific T cells in PP, and these DC act as an inducer of IL-10 producing T cells in oral tolerance.
- Published
- 2009
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160. Suppressive effects of Bifidobacterium longum on the production of Th2-attracting chemokines induced with T cell-antigen-presenting cell interactions.
- Author
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Iwabuchi N, Takahashi N, Xiao JZ, Yonezawa S, Yaeshima T, Iwatsuki K, and Hachimura S
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- Animals, Cells, Cultured, Lymph Nodes immunology, Mice, Mice, Inbred BALB C, Peyer's Patches immunology, Probiotics pharmacology, Spleen immunology, Antigen-Presenting Cells immunology, Antigen-Presenting Cells microbiology, Bifidobacterium immunology, Chemokine CCL17 antagonists & inhibitors, Chemokine CCL17 biosynthesis, Chemokine CCL22 antagonists & inhibitors, Chemokine CCL22 biosynthesis
- Abstract
In human trials, Bifidobacterium longum BB536 alleviates subjective symptoms of Japanese cedar pollinosis, an IgE-mediated type I allergy caused by exposure to Japanese cedar, and significantly suppresses the increase of plasma thymus- and activation-regulated chemokine (TARC) associated with pollen dispersion. In the present study, we investigated the suppressive effects of BB536 on the production of T helper type 2 (Th2)-attracting chemokines, such as TARC and macrophage-derived chemokine (MDC), together with the mechanisms of their production. Murine splenocytes were cultured with heat-killed BB536, and the levels of Th2-attracting chemokines in the supernatants were measured. TARC and MDC were produced in cultures without stimulation, and the production was significantly suppressed by BB536. These chemokines were produced by antigen-presenting cells (APCs) of splenocytes stimulated with an anti-CD40 antibody. Furthermore, TARC production was induced with granulocyte macrophage colony-stimulating factor that was produced by T cells and dendritic cells. BB536 suppressed MDC production induced with the anti-CD40 antibody by APCs from the spleen, mesenteric lymph nodes (MLNs) and Peyer's patches, and it suppressed TARC production by APCs from the spleen and MLNs. These results indicate that BB536 suppresses the production of Th2-attracting chemokines induced by the T cell-APC interaction, suggesting a novel mechanism for alleviating symptoms of allergic disorders by probiotics.
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- 2009
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161. Bacteroides induce higher IgA production than Lactobacillus by increasing activation-induced cytidine deaminase expression in B cells in murine Peyer's patches.
- Author
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Yanagibashi T, Hosono A, Oyama A, Tsuda M, Hachimura S, Takahashi Y, Itoh K, Hirayama K, Takahashi K, and Kaminogawa S
- Subjects
- Animals, B-Lymphocytes cytology, B-Lymphocytes immunology, B-Lymphocytes microbiology, Cell Differentiation, Coculture Techniques, Interleukin-5 metabolism, Intestinal Mucosa metabolism, Intestines cytology, Intestines immunology, Intestines microbiology, Mice, Mice, Inbred BALB C, Peyer's Patches cytology, Up-Regulation, B-Lymphocytes metabolism, Bacteroides physiology, Cytidine Deaminase genetics, Gene Expression Regulation, Enzymologic, Immunoglobulin A biosynthesis, Lactobacillus physiology, Peyer's Patches immunology
- Abstract
The gut mucosal immune system is crucial in host defense against infection by pathogenic microbacteria and viruses via the production of IgA. Previous studies have shown that intestinal commensal bacteria enhance mucosal IgA production. However, it is poorly understood how these bacteria induce IgA production and which genera of intestinal commensal bacteria induce IgA production effectively. In this study, we compared the immunomodulatory effects of Bacteroides and Lactobacillus on IgA production by Peyer's patches lymphocytes. IgA production by Peyer's patches lymphocytes co-cultured with Bacteroides was higher than with Lactobacillus. In addition, the expression of activation-induced cytidine deaminase increased in co-culture with Bacteroides but not with Lactobacillus. We found that intestinal commensal bacteria elicited IgA production. In particular, Bacteroides induced the differentiation of Peyer's patches B cell into IgA(+) B cells by increasing activation-induced cytidine deaminase expression.
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- 2009
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162. Intestinal Bifidobacterium association in germ-free T cell receptor transgenic mice down-regulates dietary antigen-specific immune responses of the small intestine but enhances those of the large intestine.
- Author
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Tsuda M, Hosono A, Yanagibashi T, Hachimura S, Hirayama K, Umesaki Y, Itoh K, Takahashi K, and Kaminogawa S
- Subjects
- Administration, Oral, Animals, Bifidobacterium metabolism, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes microbiology, CD4-Positive T-Lymphocytes pathology, Clostridium immunology, Clostridium isolation & purification, Clostridium pathogenicity, Clostridium Infections immunology, Colony Count, Microbial, Diet, Down-Regulation, Immunity, Mucosal, Interferon-gamma metabolism, Interleukin-6 metabolism, Intestine, Large microbiology, Intestine, Large pathology, Intestine, Small microbiology, Intestine, Small pathology, L-Selectin genetics, L-Selectin metabolism, Lymphocyte Activation, Mice, Mice, Transgenic, Organ Specificity, Ovalbumin administration & dosage, Ovalbumin immunology, Receptors, Antigen, T-Cell genetics, Bifidobacterium immunology, CD4-Positive T-Lymphocytes metabolism, Germ-Free Life, Intestine, Large immunology, Intestine, Small immunology
- Abstract
Bifidobacterium is a dominant bacterial species among commensals in the human intestine and is thought to have probiotic immunomodulatory effects. In this study, we investigated the effect of the association with Bifidobacterium pseudocatenulatum JCM 7041 (Bp) on dietary ovalbumin (OVA)-specific immune responses using germ-free OVA-specific T cell receptor transgenic mice (OVA23-3 mice). We established germ-free OVA23-3 mice, and then associated with Bp (BIF group) or without (CONT group) and additionally associated with segmented filamentous bacteria (SFB) and clostridia in both groups. BIF and CONT mice were fed an egg-white diet containing OVA for 1 week. Cytokine production in response to OVA by cells of Peyer's patches (PPs) and lamina propria (LP) from the small and large intestine was measured. Interferon (IFN)-gamma and interleukin (IL)-6 production by PP cells from BIF group mice was lower than that of the CONT group. The proportion of PP cells expressing CD4+CD62L(low), an activated/memory T cell phenotype, was higher in BIF group mice than the CONT group. Furthermore, LP cells from the small intestine in Bp-associated mice showed a tendency to produce slightly lower IFN-gamma and IL-6, while the cells from large intestine produced markedly higher IFN-gamma, IL-5 and IL-6 than those in the CONT group. The pattern of cytokine production by PP in BIF animals was similar to those isolated from conventional mice. These results suggest that intestinal association with Bp might down-regulate excessive immune responses to dietary antigens of the small intestine but enhance those of the large intestine.
- Published
- 2009
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163. PPARalpha gene expression is up-regulated by LXR and PXR activators in the small intestine.
- Author
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Inoue J, Satoh S, Kita M, Nakahara M, Hachimura S, Miyata M, Nishimaki-Mogami T, and Sato R
- Subjects
- Animals, Benzoates pharmacology, Benzylamines pharmacology, DNA-Binding Proteins agonists, Gene Expression drug effects, Hydrocarbons, Fluorinated, Intestine, Small metabolism, Ligands, Liver X Receptors, Male, Mice, Mice, Inbred C57BL, Orphan Nuclear Receptors, Pregnane X Receptor, Pregnenolone Carbonitrile pharmacology, RNA, Messenger metabolism, Receptors, Cytoplasmic and Nuclear agonists, Receptors, Steroid agonists, Sulfonamides pharmacology, Up-Regulation, DNA-Binding Proteins metabolism, Gene Expression Regulation, Intestine, Small drug effects, PPAR alpha genetics, Receptors, Cytoplasmic and Nuclear metabolism, Receptors, Steroid metabolism
- Abstract
LXR, PXR, and PPARalpha are members of a nuclear receptor family which regulate the expression of genes involved in lipid metabolism. Here, we show the administration of T0901317 stimulates PPARalpha gene expression in the small intestine but not in the liver of both normal and FXR-null mice. The administration of LXR specific ligand GW3965, or PXR specific ligand PCN has the same effect, indicating that ligand-dependent activation of LXR and PXR, but not FXR, is responsible for the increased gene expression of PPARalpha in the mouse small intestine.
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- 2008
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164. Growth factor-induced phosphorylation of sterol regulatory element-binding proteins inhibits sumoylation, thereby stimulating the expression of their target genes, low density lipoprotein uptake, and lipid synthesis.
- Author
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Arito M, Horiba T, Hachimura S, Inoue J, and Sato R
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- Amino Acid Substitution, Animals, Butadienes pharmacology, COS Cells, Cell Line, Tumor, Cell Nucleus genetics, Cell Nucleus metabolism, Chlorocebus aethiops, Enzyme Inhibitors pharmacology, Extracellular Signal-Regulated MAP Kinases genetics, Extracellular Signal-Regulated MAP Kinases metabolism, Farnesyl-Diphosphate Farnesyltransferase biosynthesis, Farnesyl-Diphosphate Farnesyltransferase genetics, Gene Expression Regulation drug effects, Histone Deacetylases genetics, Histone Deacetylases metabolism, Humans, Hydroxymethylglutaryl-CoA Synthase, Insulin-Like Growth Factor I pharmacology, Lipoproteins, LDL genetics, MAP Kinase Signaling System drug effects, MAP Kinase Signaling System physiology, Nitriles pharmacology, Phosphorylation drug effects, RNA, Small Interfering genetics, Receptors, LDL biosynthesis, Receptors, LDL genetics, SUMO-1 Protein genetics, Sterol Regulatory Element Binding Protein 1 genetics, Sterol Regulatory Element Binding Protein 2 genetics, Transcription, Genetic drug effects, Transcription, Genetic physiology, Ubiquitin genetics, Ubiquitin metabolism, Ubiquitination drug effects, Ubiquitination physiology, Gene Expression Regulation physiology, Insulin-Like Growth Factor I metabolism, Lipoproteins, LDL metabolism, SUMO-1 Protein metabolism, Sterol Regulatory Element Binding Protein 1 immunology, Sterol Regulatory Element Binding Protein 2 metabolism
- Abstract
The destiny and activity of sterol regulatory element-binding proteins (SREBPs) in the nucleus are regulated by modification with ubiquitin, small ubiquitin-like modifier (SUMO), or phosphorus. ERK-dependent phosphorylation causes an increase in their transcriptional activity, whereas SUMO modification halts it. We hypothesized a causal linkage between phosphorylation and sumoylation because their sites are very closely located in SREBP-1 and -2 molecules. When Ser(455), a phosphorylation site in SREBP-2, was substituted with Ala, this SREBP-2 mutant was more efficiently modified by SUMO-1. On the other hand, substitution of Asp inhibited SUMO conjugation, mimicking phosphoserine. When cells were cultured with insulin-like growth factor-1, sumoylation of SREBP-2 was decreased with an increase in its phosphorylation, but SREBP-2(S455A) was continuously sumoylated. An ERK cascade inhibitor, U0126, inversely augmented SUMO modification of SREBP-2. Insulin-like growth factor-1 treatment stimulated the expression of SREBP target genes such as the low density lipoprotein (LDL) receptor, squalene synthase, and hydroxymethylglutaryl-CoA synthase genes. These results indicate that growth factor-induced phosphorylation of SREBP-2 inhibits sumoylation, thereby facilitating SREBP transcriptional activity. Glutathione S-transferase pulldown assays revealed that wild-type SREBP-2, but not a mutant lacking Lys(464), interacts with HDAC3 preferentially among the histone deacetylase family members. HDAC3 small interfering RNA induced gene expression of the LDL receptor and thereby augmented fluorescently labeled LDL uptake in HepG2 cells. In summary, growth factors inhibit sumoylation of SREBPs through their phosphorylation, thus avoiding the recruitment of an HDAC3 corepressor complex and stimulating the lipid uptake and synthesis required for cell growth.
- Published
- 2008
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165. Lactobacillus acidophilus strain L-92 induces apoptosis of antigen-stimulated T cells by modulating dendritic cell function.
- Author
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Kanzato H, Fujiwara S, Ise W, Kaminogawa S, Sato R, and Hachimura S
- Subjects
- Animals, Antigen Presentation, Apoptosis immunology, Dendritic Cells microbiology, Female, Gram-Positive Bacterial Infections immunology, Immune System Diseases microbiology, Immune System Diseases therapy, Immunologic Memory, Lactobacillus acidophilus isolation & purification, Mice, Mice, Inbred BALB C, Mice, Transgenic, Receptors, Antigen, T-Cell immunology, Th1 Cells microbiology, Th2 Cells microbiology, Dendritic Cells immunology, Lactobacillus acidophilus immunology, Th1 Cells immunology, Th2 Cells immunology
- Abstract
Beneficial effects of lactobacilli have been reported for patients with allergic diseases and intestinal disorders such as inflammatory bowel disease. However, it is not fully understood how such bacteria influence the immunologic response. For this purpose, we investigated the effect of Lactobacillus acidophilus strain L-92 (L-92) on antigen-stimulated T cell responses in vitro and in vivo. In vitro, L-92 decreased the proliferation of CD4(+) T cells stimulated with antigen, and also induced apoptosis of antigen-stimulated T cells. On the other hand, interferon (IFN)-gamma secretion from naïve T cells was increased while interleukin (IL)-4 secretion was decreased by L-92. Co-culture with L-92 induced apoptosis of differentiated Th1 and Th2 cells. The degree of apoptosis induction was higher in Th2 cells. Moreover, L-92 up-regulated the expression of B7-H1 and down-regulated that of B7-H2 on dendritic cells (DCs), and DCs exposed to L-92 also induced apoptosis of antigen-stimulated T cells. Finally, orally administered L-92 induced apoptosis of OVA-specific TCR Tg T cells. These results indicate that L-92 attenuates the CD4(+) T cell response by inducing DC-mediated apoptosis and that it might exert beneficial effects in patients with diseases resulting from a hyper-response of CD4(+) T cells.
- Published
- 2008
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166. Prior stimulation of antigen-presenting cells with Lactobacillus regulates excessive antigen-specific cytokine responses in vitro when compared with Bacteroides.
- Author
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Tsuda M, Hosono A, Yanagibashi T, Hachimura S, Hirayama K, Itoh K, Takahashi K, and Kaminogawa S
- Abstract
The development of allergy is related to differences in the intestinal microbiota. Therefore, it is suggested that the immune responses induced by different genera of bacteria might be regulated through adaptive as well as innate immunity. In this study, we examined whether antigen-specific immune responses were affected by stimulation with the different genera of intestinal bacteria in vitro. Mesenteric lymph node (MLN) cells isolated from germ-free ovalbumin (OVA)-specific T cell receptor transgenic (OVA-Tg) mice were stimulated with OVA and intestinal bacteria. Cecal contents from conventional mice but not germ-free mice could induce OVA-specific cytokine production. Among the murine intestinal bacteria, Bacteroides acidofaciens (BA) enhanced OVA-specific IFN-gamma and IL-10 production while Lactobacillus johnsonii (LA) increased OVA-specific IL-10 production only. The expression of cell surface molecules and cytokine production by antigen-presenting cells (APCs) from germ-free Balb/c mice were analyzed. BA increased the expression of MHC II and co-stimulatory molecules on APCs compared with LA. BA increased IL-6 and IL-10 production but induced less IL-12p40 than LA. To examine the effects of prior stimulation of APCs by intestinal bacteria on the induction of antigen-specific immune responses, cytokine production was determined following co-culture with OVA, CD4(+) T cells from OVA-Tg mice, and APCs which were pre-stimulated with the bacteria or not. APCs pre-stimulated with LA did not enhance OVA-specific cytokine production while BA stimulated OVA-specific IL-10 production. These results suggest that the prior stimulation of intestinal immunocytes by Lactobacillus might regulate excessive antigen-specific cytokine responses via APCs when compared with prior stimulation by Bacteroides.
- Published
- 2007
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167. Dietary melibiose regulates th cell response and enhances the induction of oral tolerance.
- Author
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Tomita K, Nagura T, Okuhara Y, Nakajima-Adachi H, Shigematsu N, Aritsuka T, Kaminogawa S, and Hachimura S
- Subjects
- Animals, Lymphocyte Activation, Mice, Mice, Transgenic, Ovalbumin immunology, Receptors, Antigen, T-Cell, alpha-beta genetics, Th2 Cells immunology, Dietary Carbohydrates administration & dosage, Immune Tolerance, Melibiose administration & dosage, Mouth immunology, Th2 Cells drug effects
- Abstract
We examined how dietary melibiose affected the T-helper (Th) cell responses induced by an orally fed antigen in ovalbumin (OVA)-specific T cell receptor transgenic mice (OVA 23-3). Dietary melibiose markedly decreased the Th2 type responses as shown by a significant decrease in the interleukin (IL)-4 production and T cell proliferative response induced by sensitization from the 7-d oral administration of OVA. It was additionally observed that the Th1 type responses tended to decrease. We therefore examined the effect of melibiose feeding on the induction of immunological tolerance induced by the oral administration of an antigen (oral tolerance). The Th cell responses induced in BALB/c mice by subcutaneous immunization with OVA were suppressed by the prior oral administration of OVA. Such responses in the OVA-fed and immunized mice were further diminished by dietary melibiose. These results suggest that dietary melibiose strongly affected the Th cell responses to an ingested antigen, and further demonstrate the potential of melibiose to enhance the induction of oral tolerance.
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- 2007
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168. Interaction between sterol regulatory element-binding proteins and liver receptor homolog-1 reciprocally suppresses their transcriptional activities.
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Kanayama T, Arito M, So K, Hachimura S, Inoue J, and Sato R
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- Bile metabolism, Cell Line, Tumor, Cholesterol metabolism, DNA-Binding Proteins genetics, Heat-Shock Proteins metabolism, Humans, Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha, Protein Binding physiology, Protein Structure, Tertiary physiology, Receptors, Cytoplasmic and Nuclear genetics, Sterol Regulatory Element Binding Proteins genetics, Transcription Factors genetics, DNA-Binding Proteins metabolism, Gene Expression Regulation physiology, Receptors, Cytoplasmic and Nuclear metabolism, Regulatory Elements, Transcriptional physiology, Sterol Regulatory Element Binding Proteins metabolism, Transcription Factors metabolism, Transcription, Genetic physiology
- Abstract
In previous studies it was demonstrated that sterol regulatory element-binding proteins (SREBPs) are able to interact with one of the nuclear receptors, hepatocyte nuclear receptor (HNF)-4, and that this interaction regulates transcriptional activities of these proteins (Misawa, K., Horiba, T., Arimura, N., Hirano, Y., Inoue, J., Emoto, N., Shimano, H., Shimizu, M., and Sato, R. (2003) J. Biol. Chem. 278, 36176-36182; Yamamoto, T., Shimano, H., Nakagawa, Y., Ide, T., Yahagi, N., Matsuzaka, T., Nakakuki, M., Takahashi, A., Suzuki, H., Sone, H., Toyoshima, H., Sato, R., and Yamada, N. (2004) J. Biol. Chem. 279, 12027-12035). In an attempt to identify other nuclear receptor family members affecting the SREBP transcriptional activities, we found that the liver receptor homolog (LRH)-1 suppresses them. Several types of luciferase assays revealed that coexpression of these two proteins (LRH-1 and SREBP-1a, -1c, or -2) results in reciprocal inhibition of the transcriptional activity of each protein. It was confirmed that suppression in endogenous LRH-1 by small interference RNA stimulates the mRNA levels of certain SREBP target genes and that elevation in active SREBPs in the nucleus in response to cholesterol depletion suppresses the LRH-1 activity. In vitro/in vivo glutathione S-transferase pulldown experiments demonstrated that the basic helix-loop-helix-leucine zipper domain in SREBP-2 binds to the ligand-binding domain in LRH-1. Furthermore, we found that SREBP-2 interferes with the recruitment of a coactivator of LRH-1, the peroxisome proliferator-activated receptor gamma coactivator-1alpha, thereby leading to the inhibition of the LRH-1 transcriptional activity. These results clearly indicate that the interaction between SREBPs and LRH-1 exerts a suppressive influence on their target gene expression responsible for cholesterol and bile acid metabolism.
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- 2007
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169. Food antigen causes TH2-dependent enteropathy followed by tissue repair in T-cell receptor transgenic mice.
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Nakajima-Adachi H, Ebihara A, Kikuchi A, Ishida T, Sasaki K, Hirano K, Watanabe H, Asai K, Takahashi Y, Kanamori Y, Shimojo N, Matsuda H, Kohno Y, Hachimura S, and Kaminogawa S
- Subjects
- Animals, Cell Movement genetics, Cell Movement immunology, Egg Hypersensitivity genetics, Egg Hypersensitivity pathology, Egg White adverse effects, Immunoglobulin E biosynthesis, Inflammation immunology, Inflammation metabolism, Inflammation pathology, Intestinal Mucosa immunology, Intestinal Mucosa pathology, Jejunum immunology, Jejunum pathology, Male, Mast Cells immunology, Mast Cells pathology, Mice, Mice, Inbred BALB C, Mice, Transgenic, Wasting Syndrome genetics, Wasting Syndrome immunology, Wasting Syndrome pathology, Wound Healing genetics, Antigens adverse effects, Egg Hypersensitivity immunology, Intestinal Diseases immunology, Intestinal Diseases pathology, Ovalbumin adverse effects, Ovalbumin immunology, Receptors, Antigen, T-Cell genetics, Th2 Cells immunology, Wound Healing immunology
- Abstract
Background: Clarification of the mechanisms underlying the development of food-sensitive intestinal inflammation will provide an important clue to combating food allergies., Objective: To establish a model of intestinal inflammation caused by oral administration of antigen without additional treatments, we focused on the ovalbumin (OVA) 23-3 T-cell receptor transgenic mouse, which had been reported to have high serum antigen-specific IgE responses to the feeding of an egg white diet., Methods: Changes in body weight of mice fed an egg white diet were monitored throughout the 28-day experimental period. After the 28-day feeding, intestinal tissues were harvested for histologic examination. Endogenous production of cytokines and histamine in the jejunum, and production of cytokines secreted by OVA-specific CD4+ T cells purified from mesenteric lymph nodes, were analyzed., Results: Egg white diet-fed OVA23-3 mice developed weight loss and inflammation with villous atrophy and goblet cell hyperplasia, especially in the jejunum. A further characteristic feature was evidence of weight recovery and tissue repair. Jejunal inflammation was also observed in egg white diet-fed recombination activating gene (RAG)-2-deficient OVA23-3 mice. In addition, tissue sections revealed significant infiltration of specific IgE-positive cells and IgE-positive degranulating mast cells. Higher levels of IL-4 and significant levels of histamine were detected in the tissues. In the supernatant of OVA-stimulated T cells, IL-10 levels were also markedly elevated., Conclusion: We report that high-dose and continuous intake of primitive OVA alone induces enteropathy containing regions under repair in OVA23-3 mice. Antigen-specific T cells and inflammatory cells primed by T(H)2 responses play important roles in regulation of development and improvement of the disease., Clinical Implications: Long-term antigen intake causes T(H)2-dependent and food-sensitive enteropathy followed by tissue repair.
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- 2006
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170. Orally tolerized T cells can form conjugates with APCs but are defective in immunological synapse formation.
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Ise W, Nakamura K, Shimizu N, Goto H, Fujimoto K, Kaminogawa S, and Hachimura S
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- Administration, Oral, Amino Acid Sequence, Animals, Antigen-Presenting Cells enzymology, Antigen-Presenting Cells metabolism, CD4-Positive T-Lymphocytes enzymology, Cell Communication genetics, Cell Cycle Proteins metabolism, Female, Isoenzymes deficiency, Isoenzymes metabolism, Membrane Microdomains metabolism, Mice, Mice, Inbred BALB C, Mice, Transgenic, Molecular Sequence Data, Ovalbumin administration & dosage, Ovalbumin immunology, Protein Kinase C deficiency, Protein Kinase C metabolism, Protein Kinase C-theta, Protein Transport genetics, Protein Transport immunology, Proto-Oncogene Proteins deficiency, Proto-Oncogene Proteins metabolism, Proto-Oncogene Proteins c-vav, Receptors, Antigen, T-Cell deficiency, Receptors, Antigen, T-Cell genetics, Receptors, Antigen, T-Cell metabolism, cdc42 GTP-Binding Protein deficiency, cdc42 GTP-Binding Protein metabolism, rac1 GTP-Binding Protein deficiency, rac1 GTP-Binding Protein metabolism, Antigen Presentation genetics, Antigen-Presenting Cells immunology, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes transplantation, Cell Communication immunology, Immune Tolerance genetics
- Abstract
Oral tolerance is systemic immune hyporesponsiveness induced by the oral administration of soluble Ags. Hyporesponsiveness of Ag-specific CD4 T cells is responsible for this phenomenon. However, the molecular mechanisms underlying the hyporesponsive state of these T cells are not fully understood. In the present study, we investigated the ability of orally tolerized T cells to form conjugates with Ag-bearing APCs and to translocate TCR, protein kinase C-theta (PKC-theta), and lipid rafts into the interface between T cells and APCs. Orally tolerized T cells were prepared from the spleens of OVA-fed DO11.10 mice. Interestingly, the orally tolerized T cells did not show any impairment in the formation of conjugates with APCs. The conjugates were formed in a LFA-1-dependent manner. Upon antigenic stimulation, the tolerized T cells could indeed activate Rap1, which is critical for LFA-1 activation and thus cell adhesion. However, orally tolerized T cells showed defects in the translocation of TCR, PKC-theta, and lipid rafts into the interface between T cells and APCs. Translocation of TCR and PKC-theta to lipid raft fractions upon antigenic stimulation was also impaired in the tolerized T cells. Ag-induced activation of Vav, Rac1, and cdc42, which are essential for immunological synapse and raft aggregation, were down-regulated in orally tolerized T cells. These results demonstrate that orally tolerized T cells can respond to specific Ags in terms of conjugate formation but not with appropriate immunological synapse formation. This may account for the hyporesponsive state of orally tolerized T cells.
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- 2005
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171. Characteristic intestinal microflora of specific pathogen-free mice bred in two different colonies and their influence on postnatal murine immunocyte profiles.
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Nagura T, Hachimura S, Kaminogawa S, Aritsuka T, and Itoh K
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- Acetates metabolism, Animals, Cecum metabolism, Diet, Female, Hysterectomy, Lymph Nodes immunology, Mice, Oligosaccharides administration & dosage, Oligosaccharides metabolism, Pregnancy, Specific Pathogen-Free Organisms, Spleen immunology, Animals, Newborn immunology, Cecum microbiology, Lymphocyte Subsets immunology, Mice, Inbred Strains immunology, Mice, Inbred Strains microbiology
- Abstract
Cecal microflora of BALB/c mice originating from two different SPF-breeding colonies were compared. The analysis of cultivable bacteria in the ceca showed significantly higher numbers of total bacteria in BALB/cCrSlc (SLC mice) than in BALB/cA Jcl (JCL mice) (p<0.05), which were mainly based on higher numbers and occurrence of Peptococaceae. Bifidobacteria were detected only in SLC mice. Feeding an oligosaccharide, raffinose, to the mice also induced different shifts in the composition of cecal microflora and the concentration of cecal organic acids. In the second experiment, hysterectomy-derived (HD) SLC mice were fostered to SPF lactating SLC mothers, or SPF lactating JCL mice, together with the mother's own natural birth (NB) pups in each isolator. HD mice fostered to SLC-mothers showed significantly higher percentages of T-cell receptor alphabeta cells expressing a CD8alpha homodimer (p<0.05) and a CD8alphabeta heterodimer (p<0.001) in the intraepithelial lymphocytes (IEL) compared with HD mice fostered to JCL-mothers. IEL profiles of HD mice corresponded well to those of NB mice that were breast-fed by the same mothers. Differences in the ratio of B220(+)cells to Thy1.2(+)cells in the splenocytes were also observed as a trend between both HD mice fostered to SLC or JCL mothers (p=0.06). These results suggest that postnatal colonization of various characteristic intestinal microflora derived from SPF-breeding colonies results in differences in development of lymphocyte populations in the intestinal and systemic organs of mice.
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- 2005
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172. PCR method for detecting trace amounts of buckwheat (Fagopyrum spp.) in food.
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Hirao T, Imai S, Sawada H, Shiomi N, Hachimura S, and Kato H
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- DNA Primers genetics, DNA, Plant analysis, DNA, Plant genetics, Seeds genetics, Sensitivity and Specificity, Fagopyrum genetics, Food Analysis methods, Polymerase Chain Reaction methods
- Abstract
Buckwheat often causes severe allergic reactions, even when its ingestion level is extremely low. Therefore, buckwheat is listed in several countries as a common food allergen. In addition to common buckwheat and Tartarian buckwheat that are cultivated and consumed widely, wild buckwheat may be potentially allergenic. Food containing undeclared buckwheat poses a risk to patients with the buckwheat allergy. We describe in this report a PCR method to detect buckwheat DNA by using primers corresponding to the internal transcribed spacer region and the 5.8S rRNA gene. The method is buckwheat-specific and compatible with both cultivated and wild buckwheat of the Fagopyrum spp. Its sensitivity was sufficient to detect 1 ppm (w/w) of buckwheat DNA spiked in wheat DNA. This method should benefit food manufacturers, clinical doctors, and allergic patients by providing information on the presence of buckwheat contamination in food.
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- 2005
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173. T-cell receptor antagonist modifies cytokine secretion profile of naive CD4+ T cells and their differentiation into type-1 and type-2 helper T cells.
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Takato-Kaji R, Totsuka M, Ise W, Nishikawa M, Hachimura S, and Kaminogawa S
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- Amino Acid Substitution, Animals, Cell Differentiation physiology, Female, GABA Plasma Membrane Transport Proteins, Gene Expression physiology, Membrane Transport Proteins genetics, Membrane Transport Proteins metabolism, Mice, Mice, Inbred BALB C, Ovalbumin immunology, Peptide Fragments immunology, RNA, Messenger metabolism, T-Lymphocyte Subsets cytology, Th1 Cells cytology, Th2 Cells cytology, Cell Differentiation immunology, Cytokines metabolism, Receptors, Antigen, T-Cell antagonists & inhibitors, T-Lymphocyte Subsets immunology, Th1 Cells immunology, Th2 Cells immunology
- Abstract
A T-cell receptor (TCR) antagonist is an analog of a peptide ligand for TCR that inhibits T-cell responses to the original peptide. We investigated the effects of a TCR antagonist on cytokine secretion of naive CD4+ T cells and their differentiation into type-1 and type-2 helper T cells (Th1 and Th2) induced by stimulation with varying doses of an antigenic peptide. In the presence of a TCR antagonist peptide, proliferation of naive CD4+ T cells and antigen dose-dependent secretion of interferon-gamma, a typical Th1-type cytokine, by these cells was down-regulated. With respect to the secretion of interleukin-4 (IL-4), a typical Th2-type cytokine, the TCR antagonist raised the concentration of the antigenic peptide required to elicit maximal IL-4 production and, surprisingly, significantly increased the maximum level of IL-4 secretion. Similar effects induced by the TCR antagonist were observed on the Th1/Th2 differentiation of naive CD4+ T cells. These results clearly indicate that, for naive CD4+ T cells, a TCR antagonist has the potential to change the balance of Th1/Th2 cytokine secretion and even enhance Th2 responses.
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- 2005
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174. Oral antigen induces antigen-specific activation of intraepithelial CD4+ lymphocytes but suppresses their activation in spleen.
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Tamauchi H, Yoshida Y, Sato T, Hachimura S, Inoue M, Kaminogawa S, and Habu S
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- Administration, Oral, Animals, CD4-Positive T-Lymphocytes cytology, Cell Line, Tumor, Cell Proliferation, Mice, Mice, Transgenic, Spleen cytology, Xenograft Model Antitumor Assays, Antigens administration & dosage, Antigens immunology, CD4-Positive T-Lymphocytes immunology, Lymphocyte Activation, Mouth immunology, Spleen immunology
- Abstract
Intraepithelial lymphocytes (IELs) are considered to drive immune surveillance of the epithelial layer to the mucosa, which is initially exposed to exogenous antigens. However, how IELs are activated by orally administered antigens remains unclear. To clarify this mechanism, we fed ovalbumin (OVA) to T cell receptor transgenic (TCR-Tg) mice with OVA-specific MHC class II-restricted TCR and found that the cytotoxic activity of IELs was increased against both NK and LAK target cells, but notably reduced after depleting CD8 + IELs. Cytoplasmic staining showed that the production of IFN-gamma and IL-2 was increased in mice fed with OVA both in the supernatant of cultured IELs with immobilized anti-CD3 mAb and in fresh CD4+ IELs. In contrast, the cytotoxic activity against NK and LAK target cells and the production of IL-2 and IFN-gamma was decreased in splenic T cells from mice fed with OVA. However, when the splenic T cells from these mice were cultured with OVA and IL-2, IFN-gamma production recovered. The decreased response demonstrated the clonal anergy of T cells. Furthermore, tumor growth was enhanced in TCR-Tg mice carrying an OVA-transfected counterpart A20 B cell lymphoma (OVA-A20) and fed with OVA. These results indicate that the oral administration of soluble antigens can activate CD4+ IELs in an antigen-specific manner but induces hyporesponsiveness in the spleen. In addition, Th1-type cytokines produced by activated CD4+ IEL might provide a bystander effect on the cytotoxic activity of IELs.
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- 2005
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175. Hyporesponsiveness of CD4 T cells induced in oral tolerance is maintained by selective impairment in the TCR-induced calcium/NFAT signaling pathway resulting from caspase activation.
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Hachimura S, Kaji T, Asai K, Ise W, Nakayama T, and Kaminogawa S
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- Adaptor Proteins, Signal Transducing metabolism, Animals, Enzyme Activation, Lymphocyte Activation, Mice, Mice, Transgenic, NFATC Transcription Factors, Signal Transduction immunology, src Homology Domains, CD4-Positive T-Lymphocytes immunology, Calcium physiology, Caspases metabolism, DNA-Binding Proteins physiology, Immune Tolerance, Immunity, Mucosal, Nuclear Proteins physiology, Receptors, Antigen, T-Cell immunology, Signal Transduction physiology, Transcription Factors physiology
- Abstract
We examined intracellular signaling of hyporesponsive CD4 T cells induced by continuous feeding with high-dose antigen in a TCR transgenic mouse system. The results demonstrated a selective impairment in their TCR-induced calcium/NFAT-signaling pathway. Proteomic analysis revealed caspase activation in these cells, which resulted in cleavage of GADS. Further analysis of the TCR-signaling complex showed that GADS-LAT-SLP-76-associated PLC-gamma1 was decreased in both phosphorylation and association. Thus, as a consequence of caspase activation, orally tolerant CD4 T cells could not form normal TCR signaling complexes associated with GADS and showed downregulated PLC-gamma1 activation, which resulted in impairment of TCR-induced calcium signaling.
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- 2004
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176. CD4(-)c-kit(-)CD3epsilon(-)IL-2Ralpha(+) Peyer's patch cells are a novel cell subset which secrete IL-5 in response to IL-2: implications for their role in IgA production.
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Kuraoka M, Hashiguchi M, Hachimura S, and Kaminogawa S
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- Animals, CD3 Complex analysis, CD3 Complex genetics, CD4 Antigens analysis, CD4 Antigens genetics, Cells, Cultured, Dose-Response Relationship, Immunologic, Female, Flow Cytometry, Interleukin-2 Receptor alpha Subunit, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Peyer's Patches metabolism, Proto-Oncogene Proteins c-kit analysis, Proto-Oncogene Proteins c-kit genetics, Receptors, Interleukin metabolism, Immunoglobulin A biosynthesis, Interleukin-2 pharmacology, Interleukin-5 metabolism, Peyer's Patches cytology, Peyer's Patches drug effects
- Abstract
In this study, we examined which cell population contributes to IL-5 production by Peyer's patch (PP) cells. Thy1.2(-) fraction of PP cells, but not those of splenocytes, secreted IL-5 in response to IL-2. We found that CD3epsilon(-)IL-2Ralpha(+) cells purified from the Thy1.2(-)B220(-) fraction of PP cells secreted IL-5 when stimulated with IL-2. CD3epsilon(-)IL-2Ralpha(+) cells were subdivided into CD4(+) and CD4(-) populations or c-kit(+) and c-kit(-) populations, and only the CD4(-) and c-kit(-) CD3epsilon(-)IL-2Ralpha(+) cells secreted IL-5 in response to IL-2. CD3epsilon(-)IL-2Ralpha(+) cells did not express NK cell-markers and exhibited a lymphoid morphology. We have therefore identified CD3epsilon(-)IL-2Ralpha(+) cells as a unique lymphoid population that are not classified into conventional IL-5-producing cell populations, such as T cells, mast cells and NK cells. Depletion of CD3epsilon(-)IL-2Ralpha(+) cells from PP resulted in reduced IL-5 production. Furthermore, IgA secretion by B cells was increased when PP B cells were cocultured with CD3epsilon(-)IL-2Ralpha(+) cells. Taken together, these results suggest that the novel subset of CD4(-)c-kit(-)CD3epsilon(-)IL-2Ralpha(+) PP cells are capable of secreting a high level of IL-5 in response to IL-2, contribute markedly to IL-5 production and help IgA secretion by B cells.
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- 2004
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177. Vaccination with an immunodominant peptide of bovine type II collagen induces an anti-TCR response, and modulates the onset and severity of collagen-induced arthritis.
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Honda A, Ametani A, Matsumoto T, Iwaya A, Kano H, Hachimura S, Ohkawa K, Kaminogawa S, Suzuki K, Sercarz EE, and Kumar V
- Subjects
- Animals, Antibodies immunology, Arthritis, Experimental diagnosis, Binding, Competitive immunology, Cattle, Female, Histocompatibility Antigens Class II immunology, Immunosuppression Therapy, Interferon-gamma immunology, Lymphocyte Activation immunology, Mice, Mice, Inbred DBA, Peptides immunology, T-Lymphocytes immunology, Vaccination, Arthritis, Experimental immunology, Collagen Type II immunology, Immunodominant Epitopes immunology, Peptide Fragments immunology, Receptors, Antigen, T-Cell, alpha-beta immunology
- Abstract
T cell responses directed toward TCR-derived peptides have been shown to be an important regulatory mechanism of protection against autoimmunity. Here, we show that a naturally induced TCR-directed immune response can delay the onset of collagen-induced arthritis (CIA), an animal model of autoimmune rheumatoid arthritis in humans. DBA/1 mice were pretreated with an immunodominant peptide, p245-270, from bovine type II collagen (bCII) and were subsequently immunized with whole bCII for the induction of arthritis. The results showed that preactivation of p245-270-reactive cells delayed the onset and reduced the severity of CIA, compared with animals in the control group. Interestingly, the serum antibody response to bCII and the bCII-specific cytokine were not affected under these conditions. This result indicates that the observed protection was neither directly due to a lower antibody response nor due to the immune deviation of the anti-bCII T cell response. Furthermore, immunization with p245-270, but not bCII, induced a strong response to the B5 peptide, an immunodominant region of the TCR V(beta)8.2 (amino acids 76-101) that binds very strongly to I-A(q). These data suggest that at a critical phase in the loss of self-tolerance, an effective anti-TCR response, induced naturally, can regulate the pathogenic autoimmune response and thus may provide protection against autoimmunity.
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- 2004
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178. Senescence-associated decline of lymphocyte migration in gut-associated lymphoid tissues of rat small intestine.
- Author
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Ogino T, Miura S, Komoto S, Hara Y, Hokari R, Tsuzuki Y, Watanabe C, Koseki S, Nagata H, Hachimura S, Kaminogawa S, and Ishii H
- Subjects
- Adoptive Transfer, Animals, Endothelium metabolism, Immunoglobulins metabolism, Integrin alpha4 metabolism, Intestine, Small immunology, L-Selectin metabolism, Male, Mucoproteins metabolism, Peyer's Patches cytology, Rats, Rats, Inbred F344, T-Lymphocyte Subsets metabolism, Aging immunology, Cell Movement immunology, Peyer's Patches immunology, T-Lymphocyte Subsets cytology
- Abstract
Scenescence-induced changes in gut-associated lymphoid tissues may contribute largely to the impaired immune responses during aging. Age-related changes in lymphocyte recirculations were investigated in Peyer's patches of rat small intestine. Cell dynamics of labeled T lymphocytes were observed under an intravital fluorescence microscope and compared between young and aged rats. Lymphocyte transport through intestinal lymph was decreased in aged rats. The lymphocyte rolling and adherence in postcapillary venules (PCV) of Peyer's patches was also significantly impaired in aged rats, with decreased expression of L-selectin on lymphocyte surfaces. Immunohistochemical analysis revealed a significant decrease in the CD8-positive cell population in Peyer's patches of the aged group, although mucosal addressin cell adhesion molecule-1 (MAdCAM-1) expression in postcapillary venules was unaltered. Lymphocyte adoptive-transfer studies indicated that although both the donor and recipient factors influence the adherence of T cells, the former may play a predominant role in the age-related change. This study clearly demonstrated in situ that T cell migration into Peyer's patches is significantly decreased in the aged intestine, which may reflect the impaired immune responses in the aging process.
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- 2004
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179. Oral administration of food antigen induces T cell mediated intestinal inflammation: a model using TCR-transgenic mice.
- Author
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Kikuchi A, Nakajima-Adachi H, Ebihara A, Takahashi Y, Hosono A, Itoh K, Hachimura S, and Kaminogawa S
- Subjects
- Animals, Disease Models, Animal, Germ-Free Life, Intestinal Mucosa parasitology, Jejunum immunology, Jejunum pathology, Mice, Mice, Inbred BALB C, Mice, Transgenic, Ovalbumin adverse effects, Receptors, Antigen, T-Cell genetics, Weight Loss, Antigens immunology, Food adverse effects, Immunity, Mucosal, Inflammation immunology, Intestinal Mucosa immunology, Receptors, Antigen, T-Cell immunology, T-Lymphocytes immunology
- Abstract
To investigate the mechanisms inducing food-sensitive intestinal inflammation, we focused on the OVA23-3 mouse, a transgenic mouse strain expressing a T cell receptor that recognizes ovalbumin (OVA). Mice administered an egg-white (EW) diet containing OVA showed a trend of loose feces and significant weight loss. Histology of the jejunum showed severe inflammation with villous atrophy. Thus, we studied the role of T cells and intestinal microflora in the development of the inflammation. Severe villous disruption was observed in sections of the jejunum from OVA23-3 mice and RAG-2 gene-deficient OVA23-3 mice fed with EW-diet. Further, a larger number of T cells was found in the lamina propria of the jejunum of EW-diet fed OVA23-3 mice, RAG-2 gene-deficient mice and germfree OVA23-3 mice compared with those of control-diet fed mice. However, severe inflammation was not detected in the jejunum of germfree OVA23-3 mice. CD4+ T cells from the MLN of EW-diet fed OVA23-3 mice showed a Th2 cytokine secretion profile. These observations have thus clarified that antigen-specific Th2 cells play important roles in the development of intestinal inflammation. Although the presence of indigenous bacteria was not essential for the inflammation, T cells could mediate a more severe inflammatory response in their presence.
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- 2004
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180. Dendritic Cells from Spleen, Mesenteric Lymph Node and Peyer's Patch Can Induce the Production of Both IL-4 and IFN-gamma from Primary Cultures of Naive CD4(+) T Cells in a Dose-Dependent Manner.
- Author
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Hibi M, Hachimura S, Ise W, Sato A, Yoshida T, Takayama T, Sasaki K, Senga T, Hashizume S, Totsuka M, and Kaminogawa S
- Abstract
Dendritic cells (DCs) as antigen presenting cells can stimulate naive CD4(+) T cells and initiate the primary immune response which controls Th1/Th2 development. It has been suggested that DCs derived from different tissues have distinct properties. We investigated whether DCs from mesenteric lymph nodes (MLN), Peyer's patches (PP) and spleen (SPL) could induce different responses of naive CD4(+) T cells to varying doses of antigen by using a co-culture system of DCs and T cells. DCs from each tissue induced IL-4 secretion from naive CD4(+)T cells in the presence of low dose antigenic peptide, and induced IFN-gamma production at high doses of antigen. When purified CD11c(+)/B220(-) DCs were used, MLN-derived DCs induced a higher amount of IFN-gamma secretion from naive CD4(+) T cells, compared with SPL-derived DCs. We could not detect large differences in the expressions of costimulatory molecules on the surface of these two populations of DCs. On the other hand, we found that large amounts of IL-12 were secreted from MLN DCs in an antigen dose-dependent fashion. In conclusion, DCs from SPL, MLN and PP can induce the production of both IL-4 and IFN-gamma from naive CD4(+) T cells, depending on antigen dose. MLN-derived CD11c(+)/B220(-) DCs induce higher IFN-gamma production from naive CD4(+) T cells than SPL-derived DCs, through efficient IL-12 secretion.
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- 2003
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181. Interleukin 12 and CD86 Regulate Th1 and Th2 Development Induced by a Range of Antigen Doses Presented by Peyer's Patch and Spleen Cells.
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Yoshida T, Hachimura S, Ishimori M, Ise W, Totsuka M, Ametani A, and Kaminogawa S
- Abstract
In this study, we demonstrate the role of interleukin 12 (IL-12), CD80 and CD86 in T helper type 1 (Th1) and Th2 differentiation induced through antigen presentation by Peyer's patch (PP) and spleen (SPL) cells with various doses of antigen. IL-12 was found to be critical for the induction of Th1-type cytokine producing cells, while antigen-dose dependent patterns of differentiation into Th2-type cytokine producing cells were not altered by the blockade of IL-12. Further, the difference in the pattern of Th2-type cytokine producing cell differentiation induced by PP and SPL cells depending on the antigen dosage were preserved in the absence of IL-12. When the function of CD86 was blocked by specific antibody, the induction of Th1-type cytokine producing cells was kept at high levels through every antigen dose, and the difference between PP and SPL cells was abrogated. With regard to Th2 induction, CD86 enhanced the differentiation of Th2-type cytokine producing cells but it was not essential in the case of antigen presentation by SPL cells. These results suggest that antigen-dose dependent changes in Th2 cell induction are regulated by additional factors which cannot induce antigen-dose dependent changes in Th1 cell differentiation by themselves.
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- 2003
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182. Amaranth Grain Inhibits Antigen-Specific IgE Production Through Augmentation of the IFN-gamma Response in vivo and in vitro.
- Author
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Hibi M, Hachimura S, Hashizume S, Obata T, and Kaminogawa S
- Abstract
Amaranthus hypochondriacus L. (amaranth) is a nutritionally protein rich plant with a good yield, but there has been no research concerning its immunological effects in vivo or in vitro. In the present study, we examined the effects of amaranth grain on cytokine and IgE production using in vitro helper T cell development and IgE production assays and an animal model of an orally-induced, allergen-specific IgE response. First, we examined the effect of orally administered amaranth on serum IgE concentration which reflects the immune response during allergic disease. We observed significantly decreased (p < 0.05) allergen-specific IgE in the blood of mice in our animal model. We found that orally fed amaranth significantly augmented (p < 0.05) IFN-gamma production of spleen cells. In vitro studies demonstrated that the water-soluble fraction of amaranth grain promoted helper T cell type-1 (Th1) phenotype development. Moreover, we found that the amaranth grain extract suppressed antigen-specific IgE production in vitro. These data indicate that there is a component in amaranth grain which has a suppressive effect on IgE production and augments Th1 cytokine production. In conclusion, we found that amaranth grain and its extract inhibited antigen-specific IgE production through augmenting Th1 cytokine responses in vivo and in vitro.
- Published
- 2003
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183. Splenic Dendritic Cells from Antigen-Fed Mice Induced Antigen-Specific T Cell Unresponsiveness in vivo.
- Author
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Hibi M, Hachimura S, Somaya T, Toda E, Hashiguchi M, Takayama T, Sasaki K, Senga T, Hashizume S, and Kaminogawa S
- Abstract
While it is well-known that orally fed antigens induce systemic T cell tolerance (oral tolerance), the mechanism by which this occurs, however, remains unclear. In the present study, we examined the role of splenic dendritic cells (DCs) in the process of oral tolerance induction and/or maintenance, by using an adoptive transfer system of antigen (Ag)-specific CD4(+) T cells from ovalbumin (OVA)-specific T cell receptor transgenic mice and DCs from OVA-fed BALB/c mice. Transfer of splenic DCs from OVA-fed mice reduced IL-2 productivity and the proliferative activity of pre-transferred Ag-specific CD4(+) T cells to ex vivo Ag stimulation. There were no changes in expression levels of costimulatory molecules on DCs from OVA-fed mice. Our results show that orally administered Ags induce systemic T cell unresponsiveness through splenic DCs without inducing cell division of T cells, thus providing evidence that splenic DCs are involved in oral tolerance induction.
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- 2003
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184. Identification of the genes specifically expressed in orally tolerized T cells.
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Gotoh T, Ise W, Nonaka A, Hamaguchi S, Hachimura S, and Kaminogawa S
- Abstract
Oral tolerance is the systemic immunological unresponsiveness that occurs after feeding protein antigens. Its physiological role is thought to be the prevention of hypersensitivity to food antigens, and its therapeutic use to treat inflammatory diseases has been suggested. Although it has been shown that CD4(+) T cells mediate oral tolerance, the precise molecular mechanisms remain unclear. In the present study, we employed suppression subtractive hybridization and identified 10 genes specifically expressed in orally tolerized T cells. These included genes that were interesting in terms of their putative functions in the negative regulation of T cell activation, e.g. Culin 1, LAX, and Zfhx1b, as well as four genes that encoded unknown proteins. We further investigated the expression of these genes in hyporesponsive T cells induced in vitro (in vitro anergized T cells). We found that six of the 10 genes were highly expressed in these cells, and kinetic studies suggested that one was associated with the induction of anergy, while the other five were associated with the maintenance of anergy. The remaining 4 genes that were not expressed in in vitro anergized T cells are also of interest as they may play a specific role in in vivo T cell tolerance. Functional analysis of these genes should help to understand the complex mechanisms underlying the induction and maintenance of oral tolerance, and moreover, in vivo immune tolerance in general.
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- 2003
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185. CD11b+ Peyer's patch dendritic cells secrete IL-6 and induce IgA secretion from naive B cells.
- Author
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Sato A, Hashiguchi M, Toda E, Iwasaki A, Hachimura S, and Kaminogawa S
- Subjects
- Animals, Antigen Presentation, Antigen-Presenting Cells immunology, Antigen-Presenting Cells metabolism, B-Lymphocyte Subsets immunology, Cells, Cultured, Female, Immunoglobulin A, Secretory biosynthesis, Immunoglobulins biosynthesis, Interleukin-6 pharmacology, Interphase immunology, Intestinal Mucosa cytology, Intestinal Mucosa immunology, Intestinal Mucosa metabolism, Mice, Mice, Inbred BALB C, Mice, Transgenic, Peyer's Patches cytology, Spleen cytology, Spleen immunology, Spleen metabolism, Up-Regulation immunology, B-Lymphocyte Subsets metabolism, CD11b Antigen biosynthesis, Dendritic Cells immunology, Dendritic Cells metabolism, Immunoglobulin A, Secretory metabolism, Interleukin-6 metabolism, Peyer's Patches immunology, Peyer's Patches metabolism
- Abstract
Peyer's patch (PP) dendritic cells (DCs) have been shown to exhibit a distinct capacity to induce cytokine secretion from CD4(+) T cells compared with DCs in other lymphoid organs such as the spleen (SP). In this study, we investigated whether PP DCs are functionally different from DCs in the SP in their ability to induce Ab production from B cells. Compared with SP DCs, freshly isolated PP DCs induced higher levels of IgA secretion from naive B cells in DC-T cell-B cell coculture system in vitro. The IgA production induced by PP DCs was attenuated by neutralization of IL-6. In addition, the induction of IgA secretion by SP DCs, but not PP DCs, was further enhanced by the addition of exogenous IL-6. Finally, we demonstrated that only PP CD11b(+) DC subset secreted higher levels of IL-6 compared with other DC subsets in the PP and all SP DC populations, and that PP CD11b(+) DC induced naive B cells to produce higher levels of IgA compared with SP CD11b(+) DC. These results suggest a unique role of PP CD11b(+) DCs in enhancing IgA production from B cells via secretion of IL-6.
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- 2003
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186. Proteome database of unsensitized CD4 positive T lymphocytes in T cell receptor transgenic mice.
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Kaji T, Hachimura S, and Kaminogawa S
- Subjects
- Animals, CD4-Positive T-Lymphocytes immunology, Electrophoresis, Gel, Two-Dimensional, Mice, Mice, Transgenic, Proteins analysis, Receptors, Antigen, T-Cell chemistry, Receptors, Antigen, T-Cell genetics, Receptors, Antigen, T-Cell immunology, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, CD4-Positive T-Lymphocytes chemistry, Databases, Protein, Proteome
- Abstract
We established a two-dimensional electrophoresis (2-DE) mapping database of splenic CD4 T cells prepared from I-A(d)-restricted ovalbumin (OVA)(323-339) specific T cell receptor (TCR) transgenic mice (OVA23-3). First we examined the purification of CD4 T cells and found that the high purity of cells produced more accurate protein maps. The first dimension utilized narrow-range immobilized pH gradients (IPGs), pH 4.0-5.0, pH 4.5-5.5, pH 5.0-6.0, and pH 5.5-6.7. Approximately 1300 spots were detected by silver staining. Detection was performed by in-gel tryptic digestion of the spots, matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) technology and database searches via the world wide web (WWW). We have so far identified 255 proteins on 2-DE gels of whole cell lysates. This is the first construction of a proteome database for murine unsensitized CD4 T lymphocytes. To examine this further, 2-DE mapping was utilized for splenic CD4 T cells from another TCR transgenic mouse strain (DO11.10 TCR transgenic mice). Mapping patterns were found to be almost identical to those from CD4 T cells from OVA23-3 mice. These results indicated that the 2-DE maps in this study could be used for mouse CD4 T cells to examine protein changes in cells given certain stimuli.
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- 2003
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187. Proteome analysis reveals caspase activation in hyporesponsive CD4 T lymphocytes induced in vivo by the oral administration of antigen.
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Kaji T, Hachimura S, Ise W, and Kaminogawa S
- Subjects
- Animals, Apoptosis, Blotting, Western, Calcium metabolism, Carrier Proteins metabolism, Caspase 3, Cell Division, Cell Separation, DNA Fragmentation, Down-Regulation, Electrophoresis, Gel, Two-Dimensional, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Male, Mass Spectrometry, Mice, Mice, Inbred BALB C, Mice, Transgenic, Precipitin Tests, Proteins metabolism, Receptors, Antigen, T-Cell genetics, Signal Transduction, Silver Staining, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Spleen cytology, Time Factors, Up-Regulation, X-Linked Inhibitor of Apoptosis Protein, Adaptor Proteins, Signal Transducing, Administration, Oral, Antigens pharmacology, CD4-Positive T-Lymphocytes metabolism, Caspases metabolism, Enzyme Activation, Proteome
- Abstract
The oral administration of antigen can lead to systemic antigen-specific hyporesponsiveness, also known as oral tolerance. This phenomenon is a representative form of immune tolerance to exogenous antigen under physiological conditions. We have previously reported that long term feeding of dietary antigen to ovalbumin-specific T cell receptor (TCR) transgenic mice induced oral tolerance of peripheral T cells with impairment in their TCR-induced calcium-signaling pathway. In this study, we utilized two-dimensional electrophoresis to compare intracellular protein expression patterns of orally tolerant and unsensitized CD4 T cells. We detected 26 increased and 16 decreased protein spots and identified 35 of these by mass spectrometry. The results indicated that the expression of caspases was up-regulated and that the protein levels of intact proteins susceptible to caspase cleavage, such as Grb2-related adaptor downstream of Shc (GADS), were decreased in orally tolerant CD4 T cells. Western blotting experiments confirmed that expression of the active form of caspase-3 and the antiapoptotic factor, X-linked inhibitor of apoptosis, were both up-regulated in orally tolerant CD4 T cells, which were found to be nonapoptotic. We further demonstrated that orally tolerant CD4 T cells could not form normal TCR signaling complexes associated with GADS and showed down-regulated phospholipase C-gamma1 activation, which is likely to contribute to the impairment of TCR-induced calcium signaling. Our findings indicate that orally tolerant CD4 T cells up-regulate caspase activation and show decreased levels of caspase-targeted proteins, including TCR signaling-associated molecules, while up-regulating antiapoptotic factors, all of which appear to contribute to their unique tolerant characteristics.
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- 2003
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188. Antigen feeding increases frequency and antigen-specific proliferation ability of intraepithelial CD4+ T cells in alphabeta T cell receptor transgenic mice.
- Author
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Goto M, Hachimura S, Ametani A, Sato T, Kumagai Y, Habu S, Totsuka M, Ishikawa H, and Kaminogawa S
- Subjects
- Administration, Oral, Animals, Antigens immunology, Antigens pharmacology, Cell Division immunology, Cells, Cultured, Immunophenotyping, Mice, Mice, Transgenic, Ovalbumin administration & dosage, Ovalbumin immunology, Receptors, Antigen, T-Cell, alpha-beta genetics, Antigens administration & dosage, CD4-Positive T-Lymphocytes immunology, Intestinal Mucosa, Lymphocyte Activation immunology, Receptors, Antigen, T-Cell, alpha-beta immunology
- Abstract
To study how intestinal intraepithelial lymphocytes (IEL) are affected by orally ingested antigen, the phenotypes and responses of the IEL in mice expressing a transgenic T cell receptor alphabeta (TCR alphabeta) specific for ovalbumin (OVA) were analyzed after feeding OVA. In the OVA-fed mice, the abundance of alphabeta-IEL as a proportion of the total IEL population increased and the frequency of CD4+ cells increased within the TCR alphabeta+ IEL population. CD4(+) IEL from OVA-fed transgenic mice proliferated in vitro more markedly in response to antigen stimulation than IEL from mice fed the control diet. These results indicate that antigen-specific proliferation of CD4+ IEL was amplified as a result of oral administration of antigen.
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- 2003
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189. Exposure to fatty acids modulates interferon production by intraepithelial lymphocytes.
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Hara Y, Miura S, Komoto S, Inamura T, Koseki S, Watanabe C, Hokari R, Tsuzuki Y, Ogino T, Nagata H, Hachimura S, Kaminogawa S, and Ishii H
- Subjects
- Animals, Antibodies, Monoclonal pharmacology, CD3 Complex immunology, Female, Interleukin-12 pharmacology, Interleukin-18 pharmacology, Intestinal Mucosa cytology, Lymphocyte Activation, Lymphocytes cytology, Lymphocytes drug effects, Mice, Mice, Inbred BALB C, Fatty Acids pharmacology, Interferon-gamma biosynthesis, Intestinal Mucosa immunology, Lymphocytes immunology
- Abstract
Intraepithelial lymphocytes (IELs) play important roles in intestinal mucosal immunity. Although fatty acids are known to modulate the functions of immune effector cells, there has been no information about how fat exposure affects immunological function of IELs. In this study, we examined how fatty acids of various chain lengths modulate the production of interferon (IFN)-gamma by IELs stimulated with T-cell receptor (TCR) or interleukin (IL)-12/IL-18. IELs isolated from the small intestine of BALB/c mice were stimulated with plate-coated anti-CD3 monoclonal antibody (mAb) or IL-12/IL-18. They were coincubated in microtiter plates for 3 days with various concentrations of fatty acid micelles. We used arachidonic acid, linoleic acid, and oleic acid as long-chain fatty acids, and used octanoic acid as a medium-chain fatty acid. IFN-gamma in the supernatants were measured by ELISA, and the expression of IFN-gamma mRNA in IELs was determined by RT-PCR. Significant production of IFN-gamma from IELs was observed after anti-CD3 mAb stimulation. The combination of IL-12 and IL-18 induced significant levels of IFN-gamma production without TCR stimulation. Increased IFN-gamma mRNA was also observed after anti-CD3 or IL-12/IL-18 stimulation. Long-chain fatty acids dose-dependently inhibited the stimulated-IFN-gamma production at concentrations greater than 10 micro M, but the medium-chain fatty acid did not cause any significant changes in IFN-gamma production. IFN-gamma production from gammadelta IELs was very low compared with alphabeta IELs, however, both populations showed similar attenuating patterns when treated with long-chain fatty acids. There is a possibility that the exposure of IELs to intraluminal fatty acids significantly modifies the immune function of intestinal mucosa.
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- 2003
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190. CD4+CD25- T cells that express latency-associated peptide on the surface suppress CD4+CD45RBhigh-induced colitis by a TGF-beta-dependent mechanism.
- Author
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Oida T, Zhang X, Goto M, Hachimura S, Totsuka M, Kaminogawa S, and Weiner HL
- Subjects
- Animals, Antibodies analysis, Antibodies metabolism, Biotinylation, CD4-Positive T-Lymphocytes chemistry, CD4-Positive T-Lymphocytes metabolism, CD4-Positive T-Lymphocytes transplantation, Cell Membrane immunology, Cell Membrane metabolism, Cells, Cultured, Coculture Techniques, Colitis physiopathology, Cytokines biosynthesis, Disease Models, Animal, Female, Flow Cytometry, Goats, Immunophenotyping, Leukocyte Common Antigens analysis, Male, Mice, Mice, Inbred BALB C, Mice, SCID, Peptide Fragments analysis, Peptide Fragments immunology, Protein Precursors analysis, Protein Precursors immunology, Receptors, Interleukin-2 analysis, Staining and Labeling, T-Lymphocyte Subsets chemistry, T-Lymphocyte Subsets metabolism, T-Lymphocyte Subsets transplantation, Transforming Growth Factor beta1, CD4-Positive T-Lymphocytes immunology, Colitis immunology, Colitis prevention & control, Leukocyte Common Antigens biosynthesis, Peptide Fragments biosynthesis, Protein Precursors biosynthesis, Receptors, Interleukin-2 biosynthesis, T-Lymphocyte Subsets immunology, Transforming Growth Factor beta physiology
- Abstract
Murine CD4(+)CD25(+) regulatory cells have been reported to express latency-associated peptide (LAP) and TGF-beta on the surface after activation, and exert regulatory function by the membrane-bound TGF-beta in vitro. We have now found that a small population of CD4(+) T cells, both CD25(+) and CD25(-), can be stained with a goat anti-LAP polyclonal Ab without being stimulated. Virtually all these LAP(+) cells are also positive for thrombospondin, which has the ability to convert latent TGF-beta to the active form. In the CD4(+)CD45RB(high)-induced colitis model of SCID mice, regulatory activity was exhibited not only by CD25(+)LAP(+) and CD25(+)LAP(-) cells, but also by CD25(-)LAP(+) cells. CD4(+)CD25(-)LAP(+) T cells were part of the CD45RB(low) cell fraction. CD4(+)CD25(-)LAP(-)CD45RB(low) cells had minimal, if any, regulatory activity in the colitis model. The regulatory function of CD25(-)LAP(+) cells was abrogated in vivo by anti-TGF-beta mAb. These results identify a new TGF-beta-dependent regulatory CD4(+) T cell phenotype that is CD25(-) and LAP(+).
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- 2003
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191. Nucleotides enhance the secretion of interleukin 7 from primary-cultured murine intestinal epithelial cells.
- Author
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Murakami R, Yamada K, Nagafuchi S, Hachimura S, Takahashi T, Kaminogawa S, and Totsuka M
- Abstract
Our previous studies showed that dietary nucleotides fed to mice enhanced the secretion of interleukin 7 (IL-7) and transforming growth factor beta (TGF-beta) from intestinal epithelial cells (IECs). To explore whether nucleotides influence IECs directly to enhance the secretion of the cytokines or not, the effects of nucleotides added in vitro on the cytokine secretion from primary-cultured murine IECs were examined. When the mixture of nucleotide 5'-monophosphates (CMP, GMP, IMP, and UMP) or individual nucleotide 5'-monophosphates were added to the primary culture of IECs derived from BALB/c mice, the secretion of IL-7, but not that of TGF-beta, was increased significantly. Addition of nucleotides to the culture did not alter the number of the IECs. Secretion of IL-6 and granulocyte-macrophage colony-stimulating factor, which are known to be secreted from IECs, was not enhanced by the addition of nucleotides. These results demonstrate that nucleotides can affect IECs directly to enhance the secretion of IL-7, and suggest that the increased secretion of TGF-beta from IECs by dietary nucleotides was due to indirect effects of the nucleotides, which may affect intestinal microflora or cells other than IECs that in turn influence the cytokine secretion of IECs.
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- 2002
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192. Dietary nucleotides increase the mucosal IgA response and the secretion of transforming growth factor beta from intestinal epithelial cells in mice.
- Author
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Nagafuchi S, Totsuka M, Hachimura S, Goto M, Takahashi T, Yajima T, Kuwata T, and Kaminogawa S
- Abstract
We have investigated the influence of dietary nucleotides on the intestinal immune system in ovalbumin (OVA)-specific T-cell receptor (TCR) transgenic mice (OVA-TCR Tg mice). When mice were supplied with water supplemented with 2% OVA ad libitum, the faecal OVA-specific immunoglobulin A (IgA) level significantly increased in those fed a nucleotide-supplemented diet (NT(+) diet) compared with those fed a nucleotide-free control diet (NT(-) diet). In the NT(+) diet-fed mice, secretion of transforming growth factor beta (TGF-beta), which is an isotype-specific switch factor for IgA, from intestinal epithelial cells (IECs) was significantly increased. Furthermore, an increased proportion of intestinal intraepithelial lymphocytes (IELs) bearing gammadelta TCR (TCRgammadelta(+) IELs) and increased secretion from IECs of interleukin 7 (IL-7), which is essential for the development of TCRgammadelta(+) IELs, were also observed in OVA-TCR-Tg mice fed the NT(+) diet, as we previously demonstrated using BALB/c mice (Nagafuchi et al., Biosci. Biotechnol. Biochem. 64: 1459-65 (2000)). Considering that TCRgammadelta(+) T cells and TGF-beta are important for an induction of the mucosal IgA response, our results suggest that dietary nucleotides augment the mucosal OVA-specific IgA response by increasing the secretion of TGF-beta from IECs and the proportion of TCRgammadelta(+) IELs.
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- 2002
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193. Murine Peyer's patch dendritic cells prime naïve CD4(+) T cells to produce interferon-gamma.
- Author
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Sato A, Hashiguchi M, Hachimura S, and Kaminogawa S
- Abstract
We investigated the role of Peyer's patch (PP) dendritic cells (DCs) in the production of interferon (IFN)-gamma from naïve CD4(+) T cells of T cell receptor transgenic mice. PP DCs were found to prime naïve CD4(+) T cells for the production of higher levels of IFN-gamma, when compared to spleen (SP) DCs. However, a similar level of interleukin-12 (IL-12) production was observed for PP and SP DCs stimulated via the CD40 molecule. In addition, PP DCs expressed slightly higher levels of B7.2 (CD86) compared to SP DCs. This data demonstrates that PP DCs have a distinct function in the induction of IFN-gammas and suggests that PP DCs may enhance IFN-gamma production via another cytokine or costimulatory molecule, in addition to IL-12.
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- 2002
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194. T cell hyporesponsiveness induced by oral administration of ovalbumin is associated with impaired NFAT nuclear translocation and p27kip1 degradation.
- Author
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Asai K, Hachimura S, Kimura M, Toraya T, Yamashita M, Nakayama T, and Kaminogawa S
- Subjects
- Active Transport, Cell Nucleus drug effects, Active Transport, Cell Nucleus genetics, Active Transport, Cell Nucleus immunology, Administration, Oral, Amino Acid Sequence, Animals, Antibody Formation genetics, CD4-Positive T-Lymphocytes drug effects, CD4-Positive T-Lymphocytes metabolism, Calcium Signaling drug effects, Calcium Signaling genetics, Calcium Signaling immunology, Carrier Proteins metabolism, Cell Cycle genetics, Cell Cycle immunology, Cyclin-Dependent Kinase Inhibitor p27, Cyclin-Dependent Kinases antagonists & inhibitors, DNA-Binding Proteins antagonists & inhibitors, Dose-Response Relationship, Immunologic, Immediate-Early Proteins biosynthesis, Interleukin-2 pharmacology, Ionomycin pharmacology, Isoenzymes antagonists & inhibitors, Isoenzymes metabolism, MAP Kinase Signaling System genetics, MAP Kinase Signaling System immunology, Membrane Proteins metabolism, Mice, Mice, Inbred BALB C, Mice, Knockout, Mice, Transgenic, Mitogen-Activated Protein Kinase Kinases physiology, Mitogen-Activated Protein Kinases physiology, Molecular Sequence Data, NFATC Transcription Factors, Phospholipase C gamma, Phosphoproteins metabolism, Phosphorylation, Protein-Tyrosine Kinases metabolism, Receptors, Antigen, T-Cell metabolism, Receptors, Antigen, T-Cell, alpha-beta genetics, Receptors, Interleukin-2 biosynthesis, STAT5 Transcription Factor, Spleen cytology, Spleen immunology, Spleen metabolism, Suppressor of Cytokine Signaling Proteins, Trans-Activators metabolism, Transcription Factors antagonists & inhibitors, Type C Phospholipases antagonists & inhibitors, Type C Phospholipases metabolism, Tyrosine metabolism, ZAP-70 Protein-Tyrosine Kinase, Adaptor Proteins, Signal Transducing, CD4-Positive T-Lymphocytes immunology, Cell Cycle Proteins metabolism, Clonal Anergy drug effects, Clonal Anergy genetics, DNA-Binding Proteins metabolism, MAP Kinase Kinase 4, Milk Proteins, Nuclear Proteins, Ovalbumin administration & dosage, Ovalbumin immunology, Transcription Factors metabolism, Tumor Suppressor Proteins metabolism
- Abstract
Oral tolerance is an important physiological component of the immune system whereby the organism avoids dangerous reactions such as hypersensitivity to ingested food proteins and other luminal Ags which may cause tissue damage and inflammation. In addition, it has been shown in animal models and in humans that oral tolerance can be applied to controlling undesired immune responses, including autoimmune diseases, allergies, and organ transplant rejections. However, the molecular mechanisms of oral tolerance have been poorly defined. In this study, we investigated the molecular basis underlying the hyporesponsiveness of orally tolerant CD4 T cells using a TCR transgenic mouse system in which oral tolerance was induced by long-term feeding with high dose Ag. We demonstrate that the hyporesponsive state of the CD4 T cells was maintained by a selective impairment in the TCR-induced calcium/NFAT signaling pathway and in the IL-2R-induced degradation of p27(kip1) and cell cycle progression. Thus, physiological mucosal tolerance is revealed to be associated with a unique type of T cell hyporesponsiveness which differs from previously described anergic T cells.
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- 2002
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195. Suppressive effect of dietary raffinose on T-helper 2 cell-mediated immunity.
- Author
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Nagura T, Hachimura S, Hashiguchi M, Ueda Y, Kanno T, Kikuchi H, Sayama K, and Kaminogawa S
- Subjects
- Animals, CD4-Positive T-Lymphocytes immunology, Cells, Cultured, Dose-Response Relationship, Immunologic, Immunoglobulin E immunology, Interleukin-12 metabolism, Interleukin-2 metabolism, Interleukin-4 metabolism, Lymph Nodes metabolism, Lymphocyte Activation, Mice, Mice, Inbred BALB C, Mice, Transgenic, Models, Animal, Ovalbumin, Peyer's Patches metabolism, Receptors, Antigen, T-Cell metabolism, Hypersensitivity immunology, Raffinose administration & dosage, Th2 Cells immunology
- Abstract
The effects of the dietary oligosaccharide raffinose on immune responses, with special reference to its anti-allergic functions, were examined in vivo. First, feeding a diet supplemented with 50 g raffinose/kg to BALB/c mice significantly (P<0.05) increased interleukin (IL) 12 secretion from isolated Peyer's patch (PP) cells in vitro compared with feeding control diet. When isolated PP cells were used as antigen-presenting cells (APC) for CD4+ T-splenocytes isolated from ovalbumin (OVA)-specific T-cell receptor transgenic (Tg) mice in the presence of OVA as antigen, significantly (P<0.05) higher levels of interferon-gamma were observed in the cultures using APC from raffinose-fed mice than those cultures using APC from control mice. Second, the diet containing 50 g raffinose/kg or control diet was fed to OVA Tg mice, and subsequently, OVA was added to each diet to prime T cells in vivo. CD4+ T-cells from the mesenteric lymph nodes of the raffinose-fed mice secreted significantly (P<0.05) higher levels of IL-2 and significantly (P<0.05) lower levels of IL-4 following in vitro antigenic stimulation compared with those of the control mice. These present results suggest that feeding raffinose may suppress differentiation of naïve T-helper (Th) cells into Th2 cells in the mesenteric lymphoid nodes. Last, feeding raffinose suppressed rises of serum immunoglobulin E levels in the Tg mice treated with long-term ingestion of OVA. In conclusion, it is suggested that dietary raffinose suppresses serum immunoglobulin E response through suppression of Th2-type immune response against oral antigen in the lymphoid organs located in or near the intestine.
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- 2002
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196. Antigen presentation by Peyer's patch cells can induce both Th1- and Th2-type responses depending on antigen dosage, but a different cytokine response pattern from that of spleen cells.
- Author
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Yoshida T, Hachimura S, Ishimori M, Kinugasa F, Ise W, Totsuka M, Ametani A, and Kaminogawa S
- Subjects
- Animals, Antigens administration & dosage, Cell Division immunology, Dose-Response Relationship, Immunologic, Female, Mice, Mice, Inbred BALB C, Mice, Transgenic, Spleen cytology, Antigens immunology, Cytokines metabolism, Peyer's Patches immunology, Spleen immunology, Th1 Cells immunology, Th2 Cells immunology
- Abstract
The Th1 and Th2 preference induced by cells from the Peyer's patch (PP) and spleen (SPL) with various doses of an antigen was examined. The same splenic T cell receptor-transgenic CD4+ T cells were first incubated with PP or SPL cells in the presence of various doses of an antigen, and the cytokine response was observed after secondary stimulation. A Th2-type pattern was only obtained for primary stimulation at 10 microM of the antigen with PP cells, whereas a Th1 pattern was induced at both higher and lower concentrations. SPL cells in the presence of 0.1 to 1 microM of the antigen induced the secretion of Th2-type cytokines. Ten and 100 microM of the antigen plus SPL cells did not induce the release of a large quantity of cytokines. PP cells induced a different cytokine pattern at the antigen concentration that induced a similar level of T cell proliferation with SPL cells. Our findings suggest that the antigen-dose dependent development of Th1/Th2 cells is differentially modulated by the antigen-presentation function of cells in PP and SPL.
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- 2002
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197. Naive CD4+ T cells exhibit distinct expression patterns of cytokines and cell surface molecules on their primary responses to varying doses of antigen.
- Author
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Ise W, Totsuka M, Sogawa Y, Ametani A, Hachimura S, Sato T, Kumagai Y, Habu S, and Kaminogawa S
- Subjects
- Amino Acid Sequence, Animals, Antigens, Differentiation, T-Lymphocyte biosynthesis, B-Lymphocytes cytology, B-Lymphocytes immunology, CD4-Positive T-Lymphocytes cytology, CD4-Positive T-Lymphocytes metabolism, CD40 Ligand biosynthesis, Cells, Cultured, Coculture Techniques, Cytokines metabolism, Dose-Response Relationship, Immunologic, Fas Ligand Protein, Female, Ligands, Membrane Glycoproteins biosynthesis, Membrane Proteins metabolism, Mice, Mice, Inbred BALB C, Mice, Knockout, Mice, Transgenic, Molecular Sequence Data, Ovalbumin pharmacology, Peptide Fragments pharmacology, Receptors, Interleukin-4 physiology, Receptors, OX40, Signal Transduction immunology, Th2 Cells immunology, Th2 Cells metabolism, Transcription Factors biosynthesis, Tumor Necrosis Factor Receptor Superfamily, Member 7 biosynthesis, fas Receptor metabolism, Antigens pharmacology, CD4-Positive T-Lymphocytes immunology, Cytokines biosynthesis, Interphase immunology, Lymphocyte Activation immunology, Membrane Proteins biosynthesis, Ovalbumin immunology, Peptide Fragments immunology, Receptors, Tumor Necrosis Factor
- Abstract
The amount of an Ag used for stimulation affects the type and magnitude of T cell responses. In this study we have investigated the primary response of naive CD4(+) T cells derived from OVA-specific TCR-transgenic mice (OVA23-3) upon stimulation with varying doses of the antigenic peptide, OVA(323-339). IL-4 expression was maximal with 50 nM Ag and decreased significantly with increasing doses. In contrast, IFN-gamma expression, which was also detected at 50 nM Ag, increased with increasing doses. The expression patterns of mRNA for the Th2-specific transcription factors GATA-3 and c-Maf were parallel to that of IL-4. These expression profiles were not altered by the addition of anti-IL-4 plus anti-IL-12 mAbs, suggesting that cytokine receptor signaling is not essential. Naive CD4(+) T cells stimulated with 5 nM Ag elicited IgM secretion from cocultured B cells, whereas those stimulated with 50 nM Ag or more elicited apoptosis of B cells. This may be because at lower doses of Ag (5 nM), naive CD4(+) T cells express CD40 ligand and OX40, whereas at higher doses (50 nM), they express Fas ligand. Clearly, the expression of each type of molecule depends on the Ag dose, and different molecules had different expression patterns. Thus, in the primary response, naive CD4(+) T cells can exhibit different functions depending on the dose of Ag.
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- 2002
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198. Intravital observation of adhesion of lamina propria lymphocytes to microvessels of small intestine in mice.
- Author
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Fujimori H, Miura S, Koseki S, Hokari R, Komoto S, Hara Y, Hachimura S, Kaminogawa S, and Ishii H
- Subjects
- Animals, Antibodies, Monoclonal, Basement Membrane chemistry, Basement Membrane cytology, Basement Membrane immunology, Cell Adhesion immunology, Cell Adhesion Molecules, Cell Movement immunology, Female, Flow Cytometry, Immunoglobulins analysis, Immunoglobulins immunology, Immunohistochemistry, Integrins analysis, Integrins immunology, Intestinal Mucosa chemistry, L-Selectin analysis, L-Selectin immunology, Lymphatic System immunology, Lymphocyte Function-Associated Antigen-1 analysis, Lymphocyte Function-Associated Antigen-1 immunology, Mice, Mice, Inbred BALB C, Microcirculation immunology, Mucoproteins analysis, Mucoproteins immunology, Peyer's Patches blood supply, Peyer's Patches chemistry, Peyer's Patches immunology, Intestinal Mucosa blood supply, Intestinal Mucosa immunology, T-Lymphocytes cytology
- Abstract
Background & Aims: Although the recirculation of lymphocytes through the intestinal mucosa is important for the specific immune defense, the homing of lamina propria lymphocytes (LPLs) has not been clearly understood. The aim of this study is to compare, under an intravital microscope, the dynamic process of lymphocyte-endothelium recognition and binding in the murine intestinal mucosa of T lymphocytes from the lamina propria of intestine to that of T lymphocytes from the spleen., Methods: LPLs isolated from nonlymphoid areas of the small intestine and spleen (SPL) were fluorescence-labeled and injected into a jugular vein of recipient mice. Microvessels of the villus mucosa and ileal Peyer's patches were observed under an intravital fluorescence microscope, and the effects of anti-adhesion-molecule antibodies on lymphocyte-endothelial interaction were investigated., Results: LPLs accumulated abundantly in the microvessels of villus tips but not in the submucosal venules or postcapillary venules of Peyer's patches, where SPLs migrated selectively. The accumulation of LPLs in the villus tips was almost completely inhibited by anti-beta7-integrin and was significantly inhibited by anti-mucosal addressin cell-adhesion molecule 1 (MAdCAM-1) and anti-alpha4-integrin. Significant MAdCAM-1 expression was observed in the microvessels of the villus mucosa. Some SPLs adhered to the nonlymphoid mucosa, but most soon detached., Conclusions: It was shown in vivo for the first time that T lymphocytes from the lamina propria but not from the spleen adhere selectively, mostly via alpha4beta7 and MAdCAM-1, to the microvessels of villus tips of the intestine, but not to the postcapillary venules of Peyer's patches.
- Published
- 2002
- Full Text
- View/download PDF
199. Identification of multiple isolated lymphoid follicles on the antimesenteric wall of the mouse small intestine.
- Author
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Hamada H, Hiroi T, Nishiyama Y, Takahashi H, Masunaga Y, Hachimura S, Kaminogawa S, Takahashi-Iwanaga H, Iwanaga T, Kiyono H, Yamamoto H, and Ishikawa H
- Subjects
- Animals, B-Lymphocytes classification, B-Lymphocytes immunology, Flow Cytometry, Genes, Immunoglobulin, Germinal Center cytology, Germinal Center immunology, Immunity, Mucosal, Immunohistochemistry, Immunophenotyping, Intestinal Mucosa anatomy & histology, Intestinal Mucosa ultrastructure, Intestine, Small anatomy & histology, Intestine, Small ultrastructure, Leukocyte Common Antigens analysis, Leukocyte Common Antigens immunology, Mesentery immunology, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Mice, Knockout, Mice, Nude, Peyer's Patches immunology, Proto-Oncogene Proteins c-kit analysis, Proto-Oncogene Proteins c-kit immunology, Species Specificity, Intestinal Mucosa immunology, Intestine, Small immunology
- Abstract
We have revealed that 100-200 clusters, filled with closely packed lymphocytes, can be found throughout the length of the antimesenteric wall of the mouse small intestine. They are composed of a large B cell area, including a germinal center, and epithelia overlying the clusters contain M cells. A large fraction of B cells displays B220+ CD19+ CD23+ IgM(low)IgD(high)CD5(-)Mac-1(-) phenotype, and the composition of IgA+ B cells is smaller but substantial. To our knowledge, these clusters are the first identification of isolated lymphoid follicles (ILF) in mouse small intestine. ILF can be first detected at 7 (BALB/c mice) and 25 (C57BL/6 mice) days after birth, and lymphoid clusters equivalent in terms of cellular mass to ILF are present in germfree, athymic nude, RAG-2(-/-), TCR-beta(-/-), and Ig mu-chain mutant (mu(-/-)) mice, although c-kit+ cells outnumber B220+ cells in germfree and athymic nude mice, and most lymphoid residents are c-kit+ B220(-) in RAG-2(-/-), TCR-beta(-/-), and mu(-/-) mice. ILF develop normally in the progeny of transplacentally manipulated Peyer's patch (PP)-deficient mice, and decreased numbers of conspicuously atrophied ILF are present in IL-7Ralpha(-/-) PP(null) mice. Neither ILF nor PP are detectable in lymphotoxin alpha(-/-) and aly/aly mice that retain well-developed cryptopatches (CP) and thymus-independent subsets of intraepithelial T cells, whereas ILF, PP, CP, and thymus-independent subsets of intraepithelial T cells disappear from common cytokine receptor gamma-chain mutant mice. These findings indicate that ILF, PP, and CP constitute three distinct organized gut-associated lymphoid tissues that reside in the lamina propria of the mouse small intestine.
- Published
- 2002
- Full Text
- View/download PDF
200. Orally tolerant CD4 T cells respond poorly to antigenic stimulation but strongly to direct stimulation of intracellular signaling pathways.
- Author
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Asai K, Hachimura S, Toraya T, and Kaminogawa S
- Abstract
The response of splenic CD4 T cells from ovalbumin (OVA)-specific T cell receptor (TCR) transgenic mice after long-term feeding of a diet containing this antigen was examined. These CD4 T cells exhibited a decreased response to OVA peptide stimulation, in terms of proliferation, interleukin-2 secretion, and CD40 ligand expression, compared to those from mice fed a control diet lacking OVA, demonstrating that oral tolerance of T cells had been induced through oral intake of the antigen. We investigated the intracellular signaling pathways, which were Ca/CN cascade and Ras/MAPK cascade, of these tolerant CD4 T cells using phorbol-12-myristate-13-acetate (PMA) and ionomycin, which are known to directly stimulate these pathways. In contrast to the decreased response to TCR stimulation by OVA peptide, it was shown that the response of splenic CD4 T cells to these reagents in the state of oral tolerance was stronger. These results suggest that splenic CD4 T cells in the state of oral tolerance have an impairment in signaling, in which signals are not transmitted from the TCR to downstream signaling pathways, and have impairments in the vicinity of TCR.
- Published
- 2001
- Full Text
- View/download PDF
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