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1.
Biol Pharm Bull ; 42(6): 1025-1029, 2019.
Article in English | MEDLINE | ID: mdl-31155577

ABSTRACT

Magnesium oxide (MgO) is a widely used laxative. Because many antipsychotic drugs are lipophilic-basic-compounds, their solubility decreases with increasing pH and changes markedly as the pH of the solution approaches their pKa. It is highly important to clarify the effect of co-administration of MgO on the serum drug concentration for effective, safe, and appropriate medication therapy. However, the relationship between MgO administration and the serum concentration of antipsychotic drugs in patients with schizophrenia has not been reported. Therefore, in the present study, we investigated the effect of MgO administration on the concentration of antipsychotic drugs in the blood of patients with schizophrenia. The serum concentrations of biperiden, zotepine, and risperidone were assayed using an LC/MS system. The correlation between the daily dose of MgO and the relative-drug-concentration (rCp) in each patient was examined. As the MgO dose was increased, the risperidone concentration decreased. The correlation coefficient decreased for risperidone, zotepine, and biperiden, in the same order. To clarify the difference in the suppression potency of MgO on the three drugs, the relationship between the physical properties and the correlation coefficients of each drug was carefully examined. A strong correlation was observed between the pKa and the correlation coefficient. Patients with schizophrenia are often prescribed antipsychotic drugs, which have anticholinergic action and tend to suppress gastric acid secretion. We concluded that basic drug absorption might be suppressed due to an increase in the stomach pH following MgO administration. Therefore, MgO co-administration is better to avoid while taking antipsychotic drugs and anticholinergic drugs.


Subject(s)
Antipsychotic Agents/pharmacokinetics , Laxatives/pharmacology , Magnesium Oxide/pharmacology , Schizophrenia/blood , Adult , Aged , Aged, 80 and over , Antipsychotic Agents/blood , Biperiden/blood , Biperiden/pharmacokinetics , Dibenzothiepins/blood , Dibenzothiepins/pharmacokinetics , Drug Interactions , Drug Therapy, Combination , Female , Humans , Hydrophobic and Hydrophilic Interactions , Male , Middle Aged , Risperidone/blood , Risperidone/pharmacokinetics , Schizophrenia/drug therapy
2.
EBioMedicine ; 41: 584-596, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30772305

ABSTRACT

BACKGROUND: Graft-versus host disease (GVHD) is a complication of stem cell transplantation associated with significant morbidity and mortality. Non-specific immune-suppression, the mainstay of treatment, may result in immune-surveillance dysfunction and disease recurrence. METHODS: We created humanised mice model for chronic GVHD (cGVHD) by injecting cord blood (CB)-derived human CD34+CD38-CD45RA- haematopoietic stem/progenitor cells (HSPCs) into hIL-6 transgenic NOD/SCID/Il2rgKO (NSG) newborns, and compared GVHD progression with NSG newborns receiving CB CD34- cells mimicking acute GVHD. We characterised human immune cell subsets, target organ infiltration, T-cell repertoire (TCR) and transcriptome in the humanised mice. FINDINGS: In cGVHD humanised mice, we found activation of T cells in the spleen, lung, liver, and skin, activation of macrophages in lung and liver, and loss of appendages in skin, obstruction of bronchioles in lung and portal fibrosis in liver recapitulating cGVHD. Acute GVHD humanised mice showed activation of T cells with skewed TCR repertoire without significant macrophage activation. INTERPRETATION: Using humanised mouse models, we demonstrated distinct immune mechanisms contributing acute and chronic GVHD. In cGVHD model, co-activation of human HSPC-derived macrophages and T cells educated in the recipient thymus contributed to delayed onset, multi-organ disease. In acute GVHD model, mature human T cells contained in the graft resulted in rapid disease progression. These humanised mouse models may facilitate future development of new molecular medicine targeting GVHD.


Subject(s)
Graft vs Host Disease/immunology , Interleukin-6/genetics , Macrophages/immunology , T-Lymphocytes/immunology , Acute Disease , Animals , Animals, Newborn , Chronic Disease , Disease Models, Animal , Graft vs Host Disease/etiology , Graft vs Host Disease/mortality , Hematopoietic Stem Cell Transplantation , Hematopoietic Stem Cells/cytology , Hematopoietic Stem Cells/metabolism , Humans , Interleukin Receptor Common gamma Subunit/deficiency , Interleukin Receptor Common gamma Subunit/genetics , Keratinocytes/cytology , Keratinocytes/metabolism , Macrophages/metabolism , Mice , Mice, Inbred NOD , Mice, SCID , Mice, Transgenic , Survival Rate , T-Lymphocytes/metabolism , Transcriptome
3.
Immunity ; 47(2): 268-283.e9, 2017 08 15.
Article in English | MEDLINE | ID: mdl-28778586

ABSTRACT

Foxp3 controls the development and function of regulatory T (Treg) cells, but it remains elusive how Foxp3 functions in vivo. Here, we established mouse models harboring three unique missense Foxp3 mutations that were identified in patients with the autoimmune disease IPEX. The I363V and R397W mutations were loss-of-function mutations, causing multi-organ inflammation by globally compromising Treg cell physiology. By contrast, the A384T mutation induced a distinctive tissue-restricted inflammation by specifically impairing the ability of Treg cells to compete with pathogenic T cells in certain non-lymphoid tissues. Mechanistically, repressed BATF expression contributed to these A384T effects. At the molecular level, the A384T mutation altered Foxp3 interactions with its specific target genes including Batf by broadening its DNA-binding specificity. Our findings identify BATF as a critical regulator of tissue Treg cells and suggest that sequence-specific perturbations of Foxp3-DNA interactions can influence specific facets of Treg cell physiology and the immunopathologies they regulate.


Subject(s)
Basic-Leucine Zipper Transcription Factors/metabolism , Diabetes Mellitus, Type 1/congenital , Diarrhea/genetics , Forkhead Transcription Factors/metabolism , Genetic Diseases, X-Linked/genetics , Immune System Diseases/congenital , Inflammation/genetics , T-Lymphocytes, Regulatory/physiology , Alleles , Animals , Basic-Leucine Zipper Transcription Factors/genetics , Cell Differentiation , Cell Movement , Cells, Cultured , DNA Mutational Analysis , Diabetes Mellitus, Type 1/genetics , Diabetes Mellitus, Type 1/immunology , Diarrhea/immunology , Forkhead Transcription Factors/genetics , Genetic Diseases, X-Linked/immunology , Humans , Immune System Diseases/genetics , Immune System Diseases/immunology , Inflammation/immunology , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , Mutation, Missense/genetics , Organ Specificity/genetics
4.
Biopsychosoc Med ; 11: 19, 2017.
Article in English | MEDLINE | ID: mdl-28690671

ABSTRACT

BACKGROUND: Administration of valproic acid (VPA) is complicated with approximately 0.9% of patients developing hyperammonemia, but the pathogenesis of this adverse effect remains to be clarified. The aim of the present study was to search for mechanisms associated with VPA-induced hyperammonemia in the light of changes in serum amino acids concentrations associated with the urea cycle of schizophrenic patients. METHOD: Blood samples (10 mL) were obtained from 37 schizophrenic patients receiving VPA for the prevention of violent behaviors in the morning after overnight fast. Blood concentrations of ammonia, VPA, free carnitine, acyl-carnitine, and 40 amino acids including glutamate and citrulline were measured for each patient. Univariate and multivariate regression analyses were performed to identify amino acids or concomitantly administered drugs that were associated with variability in the blood concentrations of ammonia. RESULT: The blood ammonia level was positively correlated with the serum glutamate concentration (r = 0.44, p < 0.01) but negatively correlated with glutamine (r = -0.41, p = 0.01), citrulline (r = -0.42, p = 0.01), and glycine concentrations (r = -0.54, p < 0.01). It was also revealed that the concomitant administration of the mood stabilizers (p = 0.04) risperidone (p = 0.03) and blonanserin (p < 0.01) was positively associated with the elevation of the blood ammonia level. CONCLUSION: We hypothisized that VPA would elevate the blood ammonia level of schizophrenic patients. The observed changes in serum amino acids are compatible with urea cycle dysfunction, possibly due to reduced carbamoyl-phosphate synthase 1 (CPS1) activity. We conclude that VPA should be prudently prescribed to schizophrenic patients, particularly those receiving mood stabilizers or certain antipsychotics.

5.
PLoS One ; 11(12): e0167952, 2016.
Article in English | MEDLINE | ID: mdl-27973543

ABSTRACT

Skin is protected by a tough but flexible multilayered barrier and is a front line for immune responses against invading particles. For many years now, skin has been a tissue where certain vaccines are injected for the prevention of infectious disease, however, the detailed mechanisms of the skin immune response are not yet well understood. Using thin and small injection needles, we carefully injected OVA into a restricted region of mouse skin, i.e., intradermal (ID), and examined the antibody response in comparison with subcutaneous (SC) injection or epicutaneous patch administration of OVA. Epicutaneous patches induced a high IgE response against OVA, but IgG production was low. High IgG production was induced by both ID and SC injection, moreover, ID injection induced higher IgG production without any adjutants. Furthermore, OVA-specific IgE production was diminished by ID injection. We found that ID injection could efficiently stimulate skin resident DCs, drive Th1-biased conditions and diminish IgE production. The ID injection response was regulated by Langerin+ dermal DCs, because OVA was taken up mainly by these cells and, after transiently deleting them, the IgE response was no longer diminished and IgG1 production was enhanced. We also tested whether ID injection might be an effective allergy treatment by attempting to inhibit ongoing IgE production in mice with experimentally induced high serum IgE levels. Multiple ID injections of OVA were shown to prevent elevation of serum OVA-specific IgE after repeated allergen challenge. In contrast, SC OVA injection could only transiently inhibit the OVA-specific IgE production. These findings indicated that ID injection results in higher induction of antigen-specific IgG, and thus may be useful for vaccine delivery with little or no adjuvant components. Moreover, the observed diminishment of IgE and induction of Th1-biased immune responses suggest that ID may be a useful injection route for allergy immunotherapy.


Subject(s)
Allergens/administration & dosage , Hypersensitivity/immunology , Immunoglobulin E/immunology , Immunoglobulin G/immunology , Immunotherapy/methods , Animals , Dendritic Cells/cytology , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Immunohistochemistry , Injections, Intradermal , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Ovalbumin/administration & dosage , Skin/immunology , Th1 Cells/cytology , Ultrasonography , Vaccines/administration & dosage
6.
Anticancer Res ; 36(11): 5957-5963, 2016 11.
Article in English | MEDLINE | ID: mdl-27793921

ABSTRACT

BACKGROUND/AIM: To investigate bioequivalence among generic and brand-name irinotecan products. MATERIALS AND METHODS: Products of Yakult and Daiichi-Sankyo (brand-name products), Sandoz, Nippon Kayaku, Taiho, and Sawai were compared with respect to their composition and antitumor activity. RESULTS: High-performance liquid chromatography demonstrated that related substances were within the acceptable range. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay revealed significant differences in cytotoxicity for four cancer cell lines among the products. The concentration of the active compound SN-38 was highest in Yakult's product (23.82 ng/ml) and lowest in Daiichi-Sankyo's product (8.96 ng/ml). MTT assay data were correlated with the SN-38 concentration, suggesting that it influenced differences in cytocidal activity among products. However, the SN-38 concentration was far lower than that of irinotecan (20 mg/ml), suggesting a negligible clinical effect. Metabolism of irinotecan to SN-38 or open-ring forms did not differ significantly among the products. CONCLUSION: The generic products showed equivalent efficacy and safety to the brand-name products.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacokinetics , Camptothecin/analogs & derivatives , Drugs, Generic/pharmacokinetics , Camptothecin/pharmacokinetics , Cell Line, Tumor , Chromatography, High Pressure Liquid , Humans , Irinotecan , Therapeutic Equivalency
7.
J Clin Invest ; 126(6): 2064-76, 2016 06 01.
Article in English | MEDLINE | ID: mdl-27111231

ABSTRACT

Skin homeostasis is maintained by the continuous proliferation and differentiation of epidermal cells. The skin forms a strong but flexible barrier against microorganisms as well as physical and chemical insults; however, the physiological mechanisms that maintain this barrier are not fully understood. Here, we have described a mutant mouse that spontaneously develops pruritic dermatitis as the result of an initial defect in skin homeostasis that is followed by induction of a Th2-biased immune response. These mice harbor a mutation that results in a single aa substitution in the JAK1 tyrosine kinase that results in hyperactivation, thereby leading to skin serine protease overexpression and disruption of skin barrier function. Accordingly, treatment with an ointment to maintain normal skin barrier function protected mutant mice from dermatitis onset. Pharmacological inhibition of JAK1 also delayed disease onset. Together, these findings indicate that JAK1-mediated signaling cascades in skin regulate the expression of proteases associated with the maintenance of skin barrier function and demonstrate that perturbation of these pathways can lead to the development of spontaneous pruritic dermatitis.


Subject(s)
Dermatitis/etiology , Janus Kinase 1/genetics , Janus Kinase 1/metabolism , Mutant Proteins/genetics , Mutant Proteins/metabolism , Pruritus/etiology , Amino Acid Substitution , Animals , Dermatitis/enzymology , Dermatitis/genetics , Dermatitis, Atopic/enzymology , Dermatitis, Atopic/pathology , Disease Models, Animal , Enzyme Activation/genetics , Humans , Janus Kinase 1/antagonists & inhibitors , Mice , Mice, Inbred C3H , Mice, Inbred C57BL , Mice, Knockout , Mice, Mutant Strains , Mutant Proteins/antagonists & inhibitors , Mutation, Missense , Pruritus/enzymology , Pruritus/genetics , Signal Transduction , Skin/enzymology , Skin/immunology , Skin/pathology
8.
Biol Pharm Bull ; 39(3): 423-7, 2016.
Article in English | MEDLINE | ID: mdl-26934932

ABSTRACT

To provide safe dental care, it is important to minimize the pain associated with the initial injection of the local anesthetic. For this purpose, a topical anesthetic is preliminarily applied to the area where a needle will be inserted in a clinical setting. In this study, we prepared new topical anesthetic formulations with favorable intra-oral retentivity and an excellent anesthetic effect, and clinically evaluated their efficacy. We used 4% lidocaine solution as an anesthetic drug and gelatin, agar, and a food thickener as a base to prepare new topical anesthetic formulations. The subjects rested in a supine position on a chair for dental practice prior to the following experiments. Firstly, about 0.2 g of the sample was applied at a test site. One minute later, the sample was removed, and a 30 G dental injection needle was inserted into the test site. The agar/gelatin-based formulation containing gelatin of 2% and agar of 1% had a moderate solidity at 25°C and a moderate fluidity at 37°C. This formulation showed a significantly greater depth than any of the commercially available topical anesthetics. The results of the present study demonstrated that the agar/gelatin-based formulation showed an excellent analgesic effect against pain associated with needle insertion.


Subject(s)
Agar , Gelatin , Lidocaine , Pain/prevention & control , Adult , Agar/chemistry , Agar/therapeutic use , Anesthetics, Local , Dental Care , Double-Blind Method , Female , Gelatin/chemistry , Gelatin/therapeutic use , Humans , Injections , Lidocaine/chemistry , Lidocaine/therapeutic use , Male , Needles , Pain Measurement
9.
PLoS Genet ; 12(1): e1005776, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26789017

ABSTRACT

Thymic medullary regions are formed in neonatal mice as islet-like structures, which increase in size over time and eventually fuse a few weeks after birth into a continuous structure. The development of medullary thymic epithelial cells (TEC) is dependent on NF-κB associated signaling though other signaling pathways may contribute. Here, we demonstrate that Stat3-mediated signals determine medullary TEC cellularity, architectural organization and hence the size of the medulla. Deleting Stat3 expression selectively in thymic epithelia precludes the postnatal enlargement of the medulla retaining a neonatal architecture of small separate medullary islets. In contrast, loss of Stat3 expression in cortical TEC neither affects the cellularity or organization of the epithelia. Activation of Stat3 is mainly positioned downstream of EGF-R as its ablation in TEC phenocopies the loss of Stat3 expression in these cells. These results indicate that Stat3 meditated signal via EGF-R is required for the postnatal development of thymic medullary regions.


Subject(s)
Cell Differentiation/genetics , Epithelial Cells , ErbB Receptors/genetics , STAT3 Transcription Factor/biosynthesis , Animals , Embryonic Development , ErbB Receptors/biosynthesis , Flow Cytometry , Gene Expression Regulation, Developmental , Mice , STAT3 Transcription Factor/genetics , Signal Transduction , T-Lymphocytes/metabolism , Thymocytes/metabolism , Thymus Gland/growth & development , Thymus Gland/metabolism
10.
Nat Commun ; 6: 7474, 2015 Jun 17.
Article in English | MEDLINE | ID: mdl-26081938

ABSTRACT

KDEL receptors are responsible for retrotransporting endoplasmic reticulum (ER) chaperones from the Golgi complex to the ER. Here we describe a role for KDEL receptor 1 (KDELR1) that involves the regulation of integrated stress responses (ISR) in T cells. Designing and using an N-ethyl-N-nitrosourea (ENU)-mutant mouse line, T-Red (naïve T-cell reduced), we show that a point mutation in KDELR1 is responsible for the reduction in the number of naïve T cells in this model owing to an increase in ISR. Mechanistic analysis shows that KDELR1 directly regulates protein phosphatase 1 (PP1), a key phosphatase for ISR in naïve T cells. T-Red KDELR1 does not associate with PP1, resulting in reduced phosphatase activity against eIF2α and subsequent expression of stress responsive genes including the proapoptotic factor Bim. These results demonstrate that KDELR1 regulates naïve T-cell homeostasis by controlling ISR.


Subject(s)
Protein Phosphatase 1/metabolism , Receptors, Peptide/metabolism , T-Lymphocytes/physiology , Amino Acid Sequence , Animals , Apoptosis Regulatory Proteins/metabolism , Bcl-2-Like Protein 11 , Eukaryotic Initiation Factor-2/metabolism , Female , Homeostasis , Immunologic Memory , Membrane Proteins/metabolism , Mice, Inbred C3H , Mice, Inbred C57BL , Mice, Knockout , Molecular Sequence Data , Phenotype , Point Mutation , Proto-Oncogene Proteins/metabolism , Receptors, Peptide/genetics , Stress, Physiological
11.
EMBO Rep ; 15(12): 1297-304, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25378482

ABSTRACT

Intrinsic Notch signaling in intestinal epithelial cells restricts secretory cell differentiation. In gut-associated lymphoid tissue (GALT), stromal cells located beneath the follicle-associated epithelium (FAE) abundantly express the Notch ligand delta-like 1 (Dll1). Here, we show that mice lacking Rbpj-a gene encoding a transcription factor implicated in Notch signaling-in intestinal epithelial cells have defective GALT maturation. This defect can be attributed to the expansion of goblet cells, which leads to the down-regulation of CCL20 in FAE. These data demonstrate that epithelial Notch signaling maintained by stromal cells contributes to the full maturation of GALT by restricting secretory cell differentiation in FAE.


Subject(s)
Epithelial Cells/cytology , Epithelial Cells/metabolism , Lymphoid Tissue/cytology , Lymphoid Tissue/metabolism , Receptors, Notch/metabolism , Stromal Cells/cytology , Animals , Cell Differentiation/physiology , In Vitro Techniques , Intestinal Mucosa/cytology , Intestinal Mucosa/metabolism , Mice , Signal Transduction/physiology , Stromal Cells/metabolism
12.
Proc Natl Acad Sci U S A ; 111(32): 11780-5, 2014 Aug 12.
Article in English | MEDLINE | ID: mdl-25074913

ABSTRACT

The immune system is influenced by the vital zinc (Zn) status, and Zn deficiency triggers lymphopenia; however, the mechanisms underlying Zn-mediated lymphocyte maintenance remain elusive. Here we investigated ZIP10, a Zn transporter expressed in the early B-cell developmental process. Genetic ablation of Zip10 in early B-cell stages resulted in significant reductions in B-cell populations, and the inducible deletion of Zip10 in pro-B cells increased the caspase activity in parallel with a decrease in intracellular Zn levels. Similarly, the depletion of intracellular Zn by a chemical chelator resulted in spontaneous caspase activation leading to cell death. Collectively, these findings indicated that ZIP10-mediated Zn homeostasis is essential for early B-cell survival. Moreover, we found that ZIP10 expression was regulated by JAK-STAT pathways, and its expression was correlated with STAT activation in human B-cell lymphoma, indicating that the JAK-STAT-ZIP10-Zn signaling axis influences the B-cell homeostasis. Our results establish a role of ZIP10 in cell survival during early B-cell development, and underscore the importance of Zn homeostasis in immune system maintenance.


Subject(s)
Apoptosis/immunology , B-Lymphocytes/immunology , B-Lymphocytes/metabolism , Cation Transport Proteins/immunology , Zinc/metabolism , Animals , B-Lymphocytes/cytology , Caspases/metabolism , Cation Transport Proteins/deficiency , Cation Transport Proteins/genetics , Cell Differentiation , Cell Survival/immunology , Cytokines/metabolism , Homeostasis , Humans , Janus Kinases/metabolism , Lymphoma, B-Cell/immunology , Lymphoma, B-Cell/metabolism , Lymphoma, B-Cell/pathology , Lymphopenia/etiology , Lymphopenia/immunology , Lymphopenia/metabolism , Mice , Mice, Knockout , Models, Immunological , STAT Transcription Factors/metabolism , Signal Transduction , Zinc/deficiency
13.
Proc Natl Acad Sci U S A ; 111(32): 11786-91, 2014 Aug 12.
Article in English | MEDLINE | ID: mdl-25074919

ABSTRACT

The humoral immune response, also called the antibody-mediated immune response, is one of the main adaptive immune systems. The essential micronutrient zinc (Zn) is known to modulate adaptive immune responses, and dysregulated Zn homeostasis leads to immunodeficiency. However, the molecular mechanisms underlying this Zn-mediated modulation are largely unknown. Here, we show that the Zn transporter SLC39A10/ZIP10 plays an important role in B-cell antigen receptor (BCR) signal transduction. Zip10-deficiency in mature B cells attenuated both T-cell-dependent and -independent immune responses in vivo. The Zip10-deficient mature B cells proliferated poorly in response to BCR cross-linking, as a result of dysregulated BCR signaling. The perturbed signaling was found to be triggered by a reduction in CD45R phosphatase activity and consequent hyperactivation of LYN, an essential protein kinase in BCR signaling. Our data suggest that ZIP10 functions as a positive regulator of CD45R to modulate the BCR signal strength, thereby setting a threshold for BCR signaling in humoral immune responses.


Subject(s)
Cation Transport Proteins/immunology , Immunity, Humoral , Receptors, Antigen, B-Cell/metabolism , Zinc/metabolism , Adaptive Immunity , Animals , B-Lymphocytes/cytology , B-Lymphocytes/immunology , B-Lymphocytes/metabolism , Cation Transport Proteins/deficiency , Cation Transport Proteins/genetics , Cell Differentiation/immunology , Cellular Senescence/immunology , Leukocyte Common Antigens/metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Signal Transduction/immunology , T-Lymphocytes/immunology
14.
Chemotherapy ; 59(4): 260-72, 2013.
Article in English | MEDLINE | ID: mdl-24457609

ABSTRACT

BACKGROUND: Despite the potent antitumor activity of CPT-11, late-onset diarrhea owing to enterohepatic circulation of SN-38 is a critical issue. METHODS: We aimed to evaluate the inhibitory potency of gefitinib against the ABCB1- or ABCG2-mediated excretion of CPT-11 and its active metabolite SN-38 in vitro and in vivo. RESULTS: Gefitinib dose-dependently enhanced the antiproliferation activity of SN-38 in vitro by inhibiting ABCG2. The inhibitory effect of gefitinib on ABCB1 was marginal. When both CPT-11 and gefitinib were administered orally to nude mice bearing human lung cancer PC-6 cells, tumor growth was markedly suppressed. By gefitinib coadministration, the lactone forms of both CPT-11 and SN-38 in the tumor tissue increased more than 2-fold. CONCLUSIONS: Gefitinib significantly enhances the antitumor efficacy of CPT-11 and its tumor distribution in vivo. Coadministration of gefitinib may provide a new means to reduce the dose of CPT-11 and to circumvent the gastrointestinal toxicity risk.


Subject(s)
ATP-Binding Cassette Transporters/metabolism , Antineoplastic Agents/pharmacology , Camptothecin/analogs & derivatives , Neoplasm Proteins/metabolism , Quinazolines/pharmacology , ATP Binding Cassette Transporter, Subfamily B , ATP Binding Cassette Transporter, Subfamily B, Member 1/antagonists & inhibitors , ATP Binding Cassette Transporter, Subfamily B, Member 1/genetics , ATP Binding Cassette Transporter, Subfamily B, Member 1/metabolism , ATP Binding Cassette Transporter, Subfamily G, Member 2 , ATP-Binding Cassette Transporters/antagonists & inhibitors , ATP-Binding Cassette Transporters/genetics , Administration, Oral , Animals , Antineoplastic Agents/adverse effects , Antineoplastic Agents/therapeutic use , Camptothecin/adverse effects , Camptothecin/pharmacology , Camptothecin/therapeutic use , Cell Line, Tumor , Cell Survival/drug effects , Diarrhea/etiology , Drug Synergism , Drug Therapy, Combination , Female , Gefitinib , HEK293 Cells , Humans , Irinotecan , Lung Neoplasms/drug therapy , Lung Neoplasms/pathology , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Proteins/antagonists & inhibitors , Neoplasm Proteins/genetics , Quinazolines/adverse effects , Quinazolines/therapeutic use , RNA, Messenger/metabolism , Transplantation, Heterologous
15.
Int Immunol ; 24(6): 369-78, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22354913

ABSTRACT

Receptor activator of nuclear factor kappa-B ligand (RANKL) expression was examined during the development of mouse fetal peripheral lymphoid organs. A shift in the expression pattern was detected during the transition from lymphoid tissue inducer (LTi) cells to lymphoid tissue organizer (LTo) cells in the lymph node (LN) anlagen but not in the Peyer's patch anlagen. In order to understand the functional impact of these changes in the fetal expression of RANKL, the RANKL function was blocked by a blocking antibody. Excess anti-RANKL antibody was administered to pregnant mice between 13.5 and 16.5 dpc and was found to completely block LN anlagen development, suggesting that RANKL function during this period is critical for LN development. In addition, small amounts of anti-RANKL antibodies were injected directly into the amniotic space at 13.5 dpc, resulting in perturbed B-cell follicle formation and high endothelial venule differentiation after birth. These results suggest that RANKL expression on LTi cells during the early phase of LN development is critical for the development LN microarchitecture.


Subject(s)
Embryo, Mammalian/metabolism , Gene Expression Profiling , Lymph Nodes/metabolism , RANK Ligand/genetics , Animals , Antibodies/immunology , Antibodies/pharmacology , Cellular Microenvironment/drug effects , Cellular Microenvironment/genetics , Embryo, Mammalian/cytology , Embryo, Mammalian/embryology , Female , Immunohistochemistry , Lymph Nodes/cytology , Lymph Nodes/embryology , Lymphoid Tissue/cytology , Lymphoid Tissue/embryology , Lymphoid Tissue/microbiology , Male , Mice , Mice, Inbred C57BL , Organogenesis/drug effects , Organogenesis/genetics , Peyer's Patches/cytology , Peyer's Patches/embryology , Peyer's Patches/metabolism , Pregnancy , RANK Ligand/immunology , RANK Ligand/metabolism , Reverse Transcriptase Polymerase Chain Reaction
16.
PLoS Biol ; 10(2): e1001255, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22346732

ABSTRACT

There is heterogeneity in invariant natural killer T (iNKT) cells based on the expression of CD4 and the IL-17 receptor B (IL-17RB), a receptor for IL-25 which is a key factor in T(H)2 immunity. However, the development pathway and precise function of these iNKT cell subtypes remain unknown. IL-17RB⁺iNKT cells are present in the thymic CD44⁺/⁻ NK1.1⁻ population and develop normally even in the absence of IL-15, which is required for maturation and homeostasis of IL-17RB⁻iNKT cells producing IFN-γ. These results suggest that iNKT cells contain at least two subtypes, IL-17RB⁺ and IL-17RB⁻ subsets. The IL-17RB⁺iNKT subtypes can be further divided into two subtypes on the basis of CD4 expression both in the thymus and in the periphery. CD4⁺ IL-17RB⁺iNKT cells produce T(H)2 (IL-13), T(H)9 (IL-9 and IL-10), and T(H)17 (IL-17A and IL-22) cytokines in response to IL-25 in an E4BP4-dependent fashion, whereas CD4⁻ IL-17RB⁺iNKT cells are a retinoic acid receptor-related orphan receptor (ROR)γt⁺ subset producing T(H)17 cytokines upon stimulation with IL-23 in an E4BP4-independent fashion. These IL-17RB⁺iNKT cell subtypes are abundantly present in the lung in the steady state and mediate the pathogenesis in virus-induced airway hyperreactivity (AHR). In this study we demonstrated that the IL-17RB⁺iNKT cell subsets develop distinct from classical iNKT cell developmental stages in the thymus and play important roles in the pathogenesis of airway diseases.


Subject(s)
Cytokines/metabolism , Natural Killer T-Cells/physiology , T-Lymphocyte Subsets/physiology , Animals , Bronchial Hyperreactivity/immunology , Bronchial Hyperreactivity/metabolism , Bronchial Hyperreactivity/pathology , Bronchial Hyperreactivity/virology , Cells, Cultured , Flow Cytometry , Gene Expression Profiling , Immunophenotyping , Liver/pathology , Lung/immunology , Lung/pathology , Lung/virology , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , Natural Killer T-Cells/pathology , Organ Specificity , Receptors, Interleukin-17/genetics , Receptors, Interleukin-17/metabolism , Respiratory Syncytial Virus Infections/immunology , Respiratory Syncytial Virus Infections/pathology , Spleen/pathology , T-Lymphocyte Subsets/metabolism , Th17 Cells/pathology , Th17 Cells/physiology , Th2 Cells/pathology , Th2 Cells/physiology , Thymus Gland/pathology , Tissue Culture Techniques
17.
J Immunol ; 186(3): 1450-7, 2011 Feb 01.
Article in English | MEDLINE | ID: mdl-21178013

ABSTRACT

Hematopoietic lymphoid tissue inducer (LTi) cells are essential for the development of secondary lymphoid tissues including lymph nodes and Peyer's patches. Two transcription factors, the helix-loop-helix inhibitor Id2 and the retinoic acid-related orphan receptor γt (Rorγt), have been shown to be crucial for LTi cell development. However, it remains unclear how the specification of multipotent hematopoietic progenitor cells toward the LTi lineage is programmed. In this study, we report impaired lymphoid tissue organogenesis in mice in which the function of Runx1/Cbfß transcription factor complexes was attenuated by the loss of either the distal promoter-derived Runx1 or Cbfß2 variant protein. We found that LTi progenitors in fetal liver, defined previously as a lineage marker-negative α4ß7 integrin (α4ß7)(+) IL-7R α-chain (IL-7Rα)(+) population, can be subdivided into Rorγt-expressing IL-7Rα(high) cells and nonexpressing IL-7Rα(mid) cells. Whereas Id2 and Rorγt are required to direct α4ß7(+)IL-7Rα(mid) cells to become α4ß7(+)IL-7Rα(high) cells, Runx1/Cbfß2 complexes are necessary for the emergence of α4ß7(+)IL-7Rα(mid) cells. In addition, the loss of Cbfß2, but not P1-Runx1, resulted in an inefficient upregulation of Rorγt in residual α4ß7(+)IL-7Rα(+) LTi cells at anlagen. Our results thus revealed that Runx1/Cbfß2 complexes regulate the differentiation of LTi cells at two stages: an early specification of hematopoietic progenitors toward the LTi lineage and a subsequent activation of Rorγt expression at anlagen.


Subject(s)
Cell Differentiation/immunology , Core Binding Factor Alpha 2 Subunit/physiology , Core Binding Factor beta Subunit/physiology , Lymphoid Tissue/immunology , Animals , Cell Lineage/genetics , Cell Lineage/immunology , Core Binding Factor Alpha 2 Subunit/deficiency , Core Binding Factor Alpha 2 Subunit/genetics , Core Binding Factor beta Subunit/deficiency , Core Binding Factor beta Subunit/genetics , Genetic Variation/immunology , Inhibitor of Differentiation Protein 2/biosynthesis , Inhibitor of Differentiation Protein 2/genetics , Inhibitor of Differentiation Protein 2/physiology , Liver/embryology , Liver/immunology , Liver/pathology , Lymphoid Tissue/cytology , Lymphoid Tissue/embryology , Mice , Mice, Mutant Strains , Multipotent Stem Cells/cytology , Multipotent Stem Cells/immunology , Multipotent Stem Cells/metabolism , Nuclear Receptor Subfamily 1, Group F, Member 3/biosynthesis , Nuclear Receptor Subfamily 1, Group F, Member 3/genetics , Nuclear Receptor Subfamily 1, Group F, Member 3/physiology
18.
J Immunol ; 185(4): 2340-9, 2010 Aug 15.
Article in English | MEDLINE | ID: mdl-20660351

ABSTRACT

Spontaneous mutant mice that showed high levels of serum IgE and an atopic dermatitis (AD)-like skin disease were found in a colony of the KOR inbred strain that was derived from Japanese wild mice. No segregation was observed between hyper-IgE-emia and dermatitis in (BALB/c x KOR mutant) N(2) mice, suggesting that the mutation can be attributed to a single recessive locus, which we designated adjm (atopic dermatitis from Japanese mice). All four adjm congenic strains in different genetic backgrounds showed both hyper-IgE-emia and dermatitis, although the disease severity varied among strains. Linkage analysis using (BALB/c x KOR-adjm/adjm) N(2) mice restricted the potential adjm locus to the 940 kb between D10Stm216 and D10Stm238 on chromosome 10. Sequence analysis of genes located in this region revealed that the gene AI429613, which encodes the mouse homologue of the human TNFR-associated factor 3-interacting protein 2 (TRAF3IP2) protein (formerly known as NF-kappaB activator 1/connection to IkappaB kinase and stress-activated protein kinase/Jun kinase), carried a single point mutation leading to the substitution of a stop codon for glutamine at amino acid position 214. TRAF3IP2 has been shown to function as an adaptor protein in signaling pathways mediated by the TNFR superfamily members CD40 and B cell-activating factor in epithelial cells and B cells as well as in the IL-17-mediated signaling pathway. Our results suggest that malfunction of the TRAF3IP2 protein causes hyper-IgE-emia through the CD40- and B cell-activating factor-mediated pathway in B cells and causes skin inflammation through the IL-17-mediated pathway. This study demonstrates that the TRAF3IP2 protein plays an important role in AD and suggests the protein as a therapeutic target to treat AD.


Subject(s)
Adaptor Proteins, Signal Transducing/genetics , Immunoglobulin E/blood , Point Mutation , Skin Diseases/genetics , Adaptor Proteins, Signal Transducing/metabolism , Animals , Blotting, Western , Dermatitis, Atopic/blood , Dermatitis, Atopic/genetics , Dermatitis, Atopic/immunology , Female , Gene Expression , Genetic Predisposition to Disease , Interleukin-17/blood , Male , Mice , Mice, Congenic , Mice, Inbred AKR , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Mutant Strains , Pedigree , Reverse Transcriptase Polymerase Chain Reaction , Skin Diseases/blood , Skin Diseases/immunology , Survival Analysis
19.
Proc Natl Acad Sci U S A ; 107(14): 6436-41, 2010 Apr 06.
Article in English | MEDLINE | ID: mdl-20308543

ABSTRACT

During the course of inflammation and its resolution, macrophages are exposed to various cytotoxic materials, including reactive oxygen species. Thus, macrophages require a protective machinery against oxidative stress to survive at the inflammatory site. Here, we showed that xCT, a component of transport system x(c)(-), was significantly up-regulated in activated infiltrating cells, including macrophages and neutrophils at the inflammatory site. System x(c)(-) mediates the uptake of extracellular L-cystine and is consequently responsible for maintenance of intracellular glutathione levels. We established a loss-of-function mouse mutant line of xCT by N-ethyl-N-nitrosourea mutagenesis. Macrophages from xCT(mu/mu) mice showed cell death in association with the excessive release of high mobility group box chromosomal protein 1 upon stimulation with LPS, suggesting that xCT deficiency causes unremitting inflammation because of the impaired survival of activated macrophages at the inflammatory site. Subcutaneous injection of 3-methylcholanthrene (3-MCA) induced the generation of fibrosarcoma in association with inflammation. When 3-MCA was injected s.c. into mice, xCT mRNA was up-regulated in situ. In xCT(mu/mu) mice, inflammatory cytokines (such as IL-1beta and TNFalpha) were overexpressed, and the generation of 3-MCA-induced fibrosarcoma was accelerated. These results clearly indicate that the defect of the protective system against oxidative stress impaired survival of activated macrophages and subsequently enhanced tumorigenecity.


Subject(s)
Amino Acid Transport System y+/metabolism , Cell Transformation, Neoplastic/immunology , Cell Transformation, Neoplastic/metabolism , Fibrosarcoma/metabolism , Fibrosarcoma/pathology , Amino Acid Transport System y+/deficiency , Amino Acid Transport System y+/immunology , Animals , Cell Death , Cell Transformation, Neoplastic/chemically induced , Cell Transformation, Neoplastic/genetics , Fibrosarcoma/chemically induced , Fibrosarcoma/immunology , Gene Expression Regulation, Neoplastic , Interleukin-1beta/immunology , Methylcholanthrene , Mice , Mice, Knockout , Tumor Necrosis Factor-alpha/immunology , Up-Regulation
20.
Am J Pathol ; 175(6): 2257-63, 2009 Dec.
Article in English | MEDLINE | ID: mdl-19893029

ABSTRACT

Calcium-independent group VIA phospholipase A(2) (iPLA(2)beta), encoded by PLA2G6, has been shown to be involved in various physiological and pathological processes, including immunity, cell death, and cell membrane homeostasis. Mutations in the PLA2G6 gene have been recently identified in patients with infantile neuroaxonal dystrophy (INAD). Subsequently, it was reported that similar neurological impairment occurs in gene-targeted mice with a null mutation of iPLA(2)beta, whose disease onset became apparent approximately 1 to 2 years after birth. Here, we report the establishment of an improved mouse model for INAD that bears a point mutation in the ankyrin repeat domain of Pla2g6 generated by N-ethyl-N-nitrosourea mutagenesis. These mutant mice developed severe motor dysfunction, including abnormal gait and poor performance in the hanging grip test, as early as 7 to 8 weeks of age, in a manner following Mendelian law. Neuropathological examination revealed widespread formation of spheroids containing tubulovesicular membranes similar to human INAD. Molecular and biochemical analysis revealed that the mutant mice expressed Pla2g6 mRNA and protein, but the mutated Pla2g6 protein had no glycerophospholipid-catalyzing enzyme activity. Because of the significantly early onset of the disease, this mouse mutant (Pla2g6-inad) could be highly useful for further studies of pathogenesis and experimental interventions in INAD and neurodegeneration.


Subject(s)
Disease Models, Animal , Group VI Phospholipases A2/genetics , Neuroaxonal Dystrophies/genetics , Neuroaxonal Dystrophies/pathology , Age of Onset , Animals , Base Sequence , Blotting, Western , Mice , Mice, Mutant Strains , Microscopy, Electron, Transmission , Point Mutation , Polymerase Chain Reaction
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