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T Taniguchi

Publications and source records attributed to T Taniguchi.

At least 577 records · Page 32Linked to original sources

Monoclonal antibodies defining distinct epitopes of the human IL-2 receptor beta chain and their differential effects on IL-2 responses.

We have established and characterized five new monoclonal antibodies (mAbs) which specifically immunoprecipitate the human interleukin-2 receptor beta chain (IL-2R beta). One of them, TU30, recognizes the intracytoplasmic 'serine-rich region' of IL-2R beta that is critical for IL-2 signal transduction. The others, TU12, TU21, TU23 and TU25, completely inhibit IL-2 binding, as does the previously characterized TU27. However, reciprocal binding competition assays show that the epitopes recognized by the individual mAbs are different from each other. The mAbs inhibit the growth of IL-2-dependent cells. The magnitude of their inhibitory effects is dependent on not only the affinities of the mAbs for IL-2R beta but also upon the number of IL-2R alpha subunits expressed on IL-2-dependent cells. These mAbs should be useful in studying the structure and function of the IL-2R.

Animals↗

Activation of IFN-beta element by IRF-1 requires a posttranslational event in addition to IRF-1 synthesis.

Expression of the Type I IFN (i.e., IFN-alpha s and IFN-beta) genes is efficiently induced by viruses at the transcriptional level. This induction is mediated by at least two types of positive regulatory elements located in the human IFN-beta gene promoter: (1) the repeated elements which bind both the transcriptional activator IRF-1 and the repressor IRF-2 (IRF-elements; IRF-Es), and (2) the kappa B element (kappa B-E), which binds NF kappa B and is located between the IRF-Es and the TATA box. In this study we demonstrate that a promoter containing synthetic IRF-E, which displays high affinity for the IRFs can be efficiently activated by Newcastle disease virus (NDV). In contrast, such activation was either very weak or nil when cells were treated by IFN-beta or tumor necrosis factor-alpha (TNF-alpha), despite the fact they both efficiently induce de novo synthesis of the short-lived IRF-1 in L929 cells. In fact, efficient activation of the IRF-E apparently requires an event in addition to de novo IRF-1 induction, which can be elicited by NDV even in the presence of protein synthesis inhibitor, cycloheximide. Moreover, efficient activation of the IRF-E by NDV is specifically inhibited by the protein kinase inhibitor, Staurosporin. Hence our results suggest the importance of IRF-1 synthesis and post-translational modification event(s), possibly phosphorylation for the efficient activation of IRF-Es, which are otherwise under negative regulation by IRF-2.

Alkaloids↗

Molecular cloning of a porcine gene syk that encodes a 72-kDa protein-tyrosine kinase showing high susceptibility to proteolysis.

By using antibodies raised against a portion of N terminus of 40-kDa kinase (Kobayashi, T., Nakamura, S., Taniguchi, T., and Yamamura, H. (1990) Eur. J. Biochem. 188, 535-540), not only 40-kDa protein but also 72-kDa protein were detected on immunoblot analysis of porcine spleen homogenate. In splenocytes preparation, the antibodies could immunoprecipitate protein-tyrosine kinase activity of the 72-kDa protein but not detect the 40-kDa protein even on immunoblot. After incubation of crude spleen homogenate at 37 degrees C with or without various protease inhibitors, immunoblot analysis revealed proteolytic breakdown of the 72-kDa protein to 40-kDa fragment. Next, using oligonucleotides designed according to partially sequenced information of the 40-kDa kinase as a probe, we have isolated a clone containing entire coding sequence for the 40-kDa kinase from a porcine spleen cDNA library. This clone had a 1884-base-pair-long open reading frame encoding 628-amino-acid polypeptide with a calculated molecular weight of 71,618. The deduced amino acid sequence did not contain a ligand binding or membrane spanning region but did a well-conserved protein-tyrosine kinase domain and two src homology region 2 domains. The sequences of these domains showed 30-40% identity to those of other protein-tyrosine kinases, but those of remaining parts were quite unique. From these results, we concluded that the 40-kDa kinase was generated by proteolysis from the 72-kDa holoprotein which was a new member of nonreceptor protein-tyrosine kinase so far reported. We therefore designated this gene as syk after spleen tyrosine kinase.

Amino Acid Sequence↗

Interaction of the IL-2 receptor with the src-family kinase p56lck: identification of novel intermolecular association.

In the interleukin-2 (IL-2) system, intracellular signal transduction is triggered by the beta chain of the IL-2 receptor (IL-2R beta); however, the responsible signaling mechanism remains unidentified. Evidence for the formation of a stable complex of IL-2R beta and the lymphocyte-specific protein tyrosine kinase p56lck is presented. Specific association sites were identified in the tyrosine kinase catalytic domain of p56lck and in the cytoplasmic domain of IL-2R beta. As a result of interaction, IL-2R beta became phosphorylated in vitro by p56lck. Treatment of T lymphocytes with IL-2 promotes p56lck kinase activity. These data suggest the participation of p56lck as a critical signaling molecule downstream of IL-2R via a novel interaction.

Adult↗

Time course of in vitro expression of NADPH-diaphorase in cultured rat brain neurons: comparison with in vivo expression.

The time course of NADPH-diaphorase expression was examined in primary cultures of rat central nervous system and in embryonic or neonatal rat brains using a histochemical method. In cerebral and brainstem cultures from 17-day-old embryonic rats, neuronal cells moderately expressing NADPH-diaphorase were first detected on about the 5th to 7th day of culturing. Both the density of positive cells and the staining intensity increased with age of cultures. The density of positive cells, calculated as a percent of the total number of cells, increased up to day 21 in cultures from both the cerebrum and the brainstem, indicating that NADPH-diaphorase is preferentially expressed in neurons with longer viability. On the other hand, virtually no intensely positive cells were detectable in cerebellar cultures at any period examined up to 21 days. In the in vivo study, moderately stained NADPH-diaphorase-positive neurons were first detected, mainly in the laterodorsal-pedunculopontine tegmental nuclei complex and partly in the striatum, in 16-day-old embryonic rat brain. At 2 days postnatal, intensely stained neurons were detectable in the cerebral cortex as well as in the tegmental nuclei complex and the striatum, indicating some delay in the in vitro, as compared to the in vivo, expression of neuronal NADPH-diaphorase.

Animals↗

Fluctuation of gene expression for poly(ADP-ribose) synthetase during hemin-induced erythroid differentiation of human leukemia K562 cells and its reversion process.

We have studied the regulation of gene expression for poly(ADP-ribose) synthetase during erythroid differentiation and its reversion process. When human leukemia K562 cells were incubated in the presence of 80 microM hemin, benzidine-positive cells appeared at day 2 and 90% of the cells became positive at day 6. However, RNA blot analysis reveals that mRNA for gamma-globin was already abundant in untreated K562 cells and the level of the message was slightly increased by hemin-treatment. Spectroscopic analysis and polyacrylamide gel electrophoresis of the induced cell extracts indicate that hemoglobin molecules were not detected in untreated cells, and increased successively up to day 6. The hemin-induced cells were thoroughly washed, and then recultured in the absence of hemin. The benzidine-positive cells mostly disappeared 3 days after the elimination of the inducer. During the hemin-induced erythroid differentiation, the activity and mRNA for poly(ADP-ribose) synthetase decreased to 50% and 20% of the initial level at day 3 and a low level of the gene expression was maintained afterwards, whereas the activity and mRNA returned to the initial value 1 day after hemin elimination. The results indicate that the hemin-induced erythroid differentiation of K562 cells is a reversible process and depression of the synthetase may be involved in the progress of differentiation.

Blotting, Northern↗

Inhibitory effect of interferon-gamma-dependent induction of major histocompatibility complex class II antigen by expressing exogenous poly(ADP-ribose) synthetase gene.

Ia antigen, one of class II major histocompatibility complexes, was induced by treatment of mouse macrophage P388D1 cells with interferon-gamma. During the process, expression of poly(ADP-ribose) synthetase gene was depressed. We examined whether or not down-regulation of the synthetase was required for the interferon-gamma-dependent induction of Ia antigen. We constructed expression plasmid harboring metallothionein promoter-regulated synthetase gene. The enzyme activity and mRNA level of the synthetase were increased in exogenous synthetase gene-transfected clones. Induction of Ia antigen gene was strongly inhibited by expressing the exogenous synthetase gene upon treatment of the transfected clones with interferon-gamma. The result suggests that interferon-gamma-dependent down-regulation of the synthetase may be involved in a step of the signal transduction to induce Ia antigen.

Animals↗

The lectin wheat germ agglutinin induces rapid protein-tyrosine phosphorylation in human platelets.

In response to wheat germ agglutinin (WGA), platelet aggregation and stimulation of protein-tyrosine phosphorylation were observed in a dose dependent manner. These reactions were completely inhibited by coexistence of N-acetyl-D-glucosamine with WGA. Upon stimulation by this agonist, protein-tyrosine phosphorylation of seven bands with molecular masses of 140-, 130-, 80-, 76-, 53-, 38- and 35-kDa proteins was observed by immunoblot. These protein-tyrosine phosphorylations were divided into three groups by kinetics. Considering the previous report from our laboratory that thrombin and collagen induced tyrosine phosphorylation in 135-, 124- and 76-kDa proteins (Nakamura, S. and Yamamura, H. (1989) J. Biol. Chem. 264, 7089-7091.), there may be another signal transduction pathway in tyrosine phosphorylation of human platelets.

Blood Platelets↗

Ionophore A23187-induced protein-tyrosine phosphorylation of human platelets: possible synergism between Ca2+ mobilization and protein kinase C activation.

Addition of ionophore A23187 to washed human platelets caused a time- and dose-dependent increase in the phosphotyrosyl content of 135, 124 and 76 kDa proteins. Platelets loaded with intracellular Ca2+ chelator 5,5'-dimethyl-bis-(o-aminophenoxy)-ethane-N, N, N', N'-tetraacetic acid before addition of A23187 exhibited no protein-tyrosine phosphorylation. Replenishment of such platelets with extracellular CaCl2 restored A23187-induced protein-tyrosine phosphorylation. Upon stimulation with A23187, both aspirin and ADP scavengers-treated platelets exhibited protein-tyrosine phosphorylation without phosphoinositide hydrolysis and protein kinase C activation. These data show (a) that A23187 stimulates protein-tyrosine phosphorylation by the elevation of intracellular Ca2+, and (b) that A23187-induced protein-tyrosine phosphorylation is independent of formation of endoperoxides/thromboxane A2, released ADP, phosphoinositide hydrolysis and protein kinase C activation. Furthermore, a synergistic effect of A23187 and protein kinase C activators in stimulating protein-tyrosine phosphorylation is suggested.

Blood Platelets↗

Requirement of down-regulation of NAD+ ADP-ribosyltransferase for the interferon-gamma-induced activation process of murine macrophage tumor cells.

Expression of the NAD+ ADP-ribosyltransferase gene is depressed during interferon-gamma-induced activation of murine macrophage P388D1 tumor cells [Taniguchi, T., Yamauchi, K., Yamamoto, T., Tokushima, K., Harada, N., Tanaka, H., Takahashi, S., Yamamoto, H. & Fujimoto, S. (1988) Eur. J. Biochem. 171, 571-575]. In order to study the role(s) of NAD+ ADP-ribosyltransferase in interferon-gamma-induced activation of P388D1 cells, we transfected an cloned synthetase gene into P388D1 cells and examined the effect of exogenous transferase gene expression on the induction of the Ia antigen, one of the major histocompatibility gene products, by interferon-gamma. The transferase activity of the transfected cells was twice that of control cells, and Southern blot analysis revealed that characteristic restriction sizes of cDNA were detected in the clones. RNA blot analysis using a cDNA for the transferase as a probe showed that the level of mRNA for the transferase in transfected cells was higher than that in control cells, and mRNA for the exogenous transferase was still detectable 2 days after the transfected cells were treated with interferon-gamma. This indicates that the exogenous transferase gene was expressed in transfected cells. RNA blot analysis with a cDNA for the Ia antigen and flow-cytometric analysis showed that the Ia antigen was induced much less in the transfected cells by interferon-gamma, in terms of the mRNA and the Ia antigen. The results suggest that down-regulation of the transferase is required for the induction of the Ia antigen in P388D1 cells by interferon-gamma.

Animals↗

Specific depletion of the B-cell population induced by aberrant expression of human interferon regulatory factor 1 gene in transgenic mice.

Interferons (IFNs) are well known both as antiviral proteins and as potent regulators of cell growth and differentiation. In fact, IFNs inhibit growth of various normal and transformed cell types. Previously, a nuclear factor, IRF-1 (interferon regulatory factor 1), which binds to type I IFN and some IFN-inducible gene promoters, was identified and cloned. Since the IRF-1 gene is both virus and IFN inducible, an intriguing issue is raised as to whether the IRF-1 gene is functioning in IFN-mediated regulation of cell growth and differentiation. In this study, we generated transgenic mice carrying the human IRF-1 gene linked to the human immunoglobulin heavy-chain enhancer. In the transgenic mice, all the lymphoid tissues examined showed a dramatic reduction in the number of B lymphocytes (B cells). Preparation and analysis of bone marrow cells from the chimeric mice indicated that the bone marrow is the effective site for specific depletion of the B-cell population. In fact, transgenic bone marrow cells cocultured with a bone marrow-derived stromal cell line revealed an altered B-cell maturation pattern.

Animals↗

Enhanced expression of poly(ADP-ribose) synthetase gene in malignant lymphoma.

Over-expression of cellular protooncogenes has been proposed to function in the initiation and maintenance of malignancies. In order to distinguish malignant lymphoma from reactive proliferative diseases, we surveyed the expression levels of three protooncogenes(c-myc, c-fos and c-myb) in malignant lymphoma and reactive proliferative diseases. An increased level of c-myc or c-fos mRNA was observed in one case, respectively, out of three malignant lymphomata. The other cases exhibited no enhancement in protooncogenes. These oncogenes are critically regulated during differentiation, but the half-life of c-myc mRNA was very short, and the level of the mRNA decreased to the initial level very quickly. Thus, the high level of the expression of these oncogenes may not always be maintained in all malignant cells. We then examined the level of mRNA for poly(ADP-ribose) synthetase in those cases. An enhanced expression for the synthetase gene was observed in all five malignant lymphomata tested, but no increase in the level of the mRNA was observed in any reactive proliferative cases or normal lymph nodes. These results suggest that enhanced expression of poly(ADP-ribose) synthetase gene seems to be a common characteristic of protopathic malignant lymphoma. By using the characteristics of malignant lymphoma, the level of mRNA for the synthetase may be applicable for differential diagnosis of malignant lymphoma from several pathologically indistinguishable diseases.

Blotting, Northern↗

The integrity of the conserved 'WS motif' common to IL-2 and other cytokine receptors is essential for ligand binding and signal transduction.

Recent studies have identified a new family of cytokine receptors, which is primarily characterized by the conservation of periodically interspersed four cysteine residues and the W-S-X-W-S sequence ('WS motif') within the extracellular domain. However, the role of such conserved structures still remains elusive, in particular that of the WS motif. Interleukin-2 (IL-2) is known to play a critical role in the clonal expansion of antigen-stimulated T lymphocytes, and the IL-2 signal is delivered by one of the receptor components, the IL-2 receptor beta (IL-2R beta) chain. The IL-2R beta chain, unlike the IL-2R alpha chain, belongs to this receptor family. In the present study, we analyzed the function of the WS motif of IL-2R beta (Trp194-Ser195-Pro196-Trp197-Ser198) with the use of site-directed mutagenesis. Our results indicate the critical role of the two Trp residues in the proper folding of the IL-2R beta extracellular domain and point to the general functional importance of the WS motif in the new cytokine receptor family.

Amino Acid Sequence↗

Expression of protein kinase C isozymes in primary neuronal cultures of the rat cerebellum.

Protein kinase C (PKC), a family of closely related enzymes, has been implicated in molecular processes involved in differentiation in a variety of cells, including neuronal cells. We studied the presence and distribution of four PKC isozymes immunocytochemically in primary neuronal cultures of the rat cerebellum. We employed four anti-PKC antisera raised against synthetic peptides predicted from the cDNA sequence of the C-terminal portion of four PKC isozymes, alpha, beta I, beta II, and gamma. The majority of neurons were PKC(beta II) immunoreactive both in the early and late (14 days) stage of culture, whereas PKC(alpha)-, (beta I)-, and (gamma)-immunoreactive neurons were most abundant in the late stage of culture. Immunoreactivity of each PKC was high in the cytoplasm, processes, and growth cones. Prominent nuclear staining was observed with anti-PKC(gamma) antibody. These results are in contrast with in vivo results where each PKC isozyme is localized in a distinct population of neurons and subcellular compartment, suggesting the presence of regulatory mechanisms for PKC expression and compartmentalization in vivo.

Animals↗

Two cases of retained foreign bodies after cholecystectomy: diagnosis by sonography, CT, angiography, and MRI.

We encountered two cases of subphrenic abscesses around surgical sponges that had been left at cholecystectomy. These cases had been operated on either eight years or seven years earlier, respectively. These lesions resembled another diseases, especially, liver tumors. In order to make a differential diagnosis, various diagnostic procedures were carried out. Sonography showed a cystic mass with spongiform structures in one case. But in another case, no information of internal structures could be obtained because of the presence of a calcified wall. On CT, the lesions appeared as either round or elliptic masses that contained calcified foci in both cases. Angiography showed the extended and compressed branches of the hepatic artery. Spotty stains were also observed in one case. This appearance resembled a hepatic tumor. MR demonstrated both T1 and T2 elongated masses. The internal structures of the mass were clearly revealed by the improved version of the MR machine. MR proved to be extremely useful in making a differential diagnosis. It is, however, important to evaluate the findings of these imaging procedures synthetically and in association with a careful review of a patient's prior surgical history.

Aged↗

An application of recombinant human granulocyte colony-stimulating factor (rhG-CSF) in a case of hepatocellular carcinoma combined with liver cirrhosis in which leukopenia developed after chemoembolization.

A 63-year-old Japanese woman who was being treated for liver cirrhosis was diagnosed as having hepatocellular carcinoma in the caudate lobe of the liver. Transcatheter hepatic arterial chemoembolization was performed for this lesion, but severe neutropenia occurred. To restore white blood cell (WBC) counts, recombinant human granulocyte colony-stimulating factor (rhG-CSF) was administered (250 micrograms per day during 10 days, intravenously). Subsequently, WBC counts recovered immediately without side effects. This suggests that rhG-CSF could be useful for the treatment of neutropenia after chemoembolization, even in cirrhotic patients.

Antineoplastic Combined Chemotherapy Protocols↗

The interferon family stimulates the secretions of prolactin and interleukin-6 by the pituitary gland in vitro.

The effects of interferon-alpha, interferon-beta 1 and interferon-gamma on the secretions of prolactin (PRL) and interleukin-6 by primary cultured rat anterior pituitary cells were examined. These three interferons caused dose-dependent increases in PRL secretion within 30 min, and dose-dependent stimulation of interleukin-6 were weaker than the effects of interleukin-1 and tumor necrosis factor-alpha. These results suggest that interferons regulate PRL secretion from the pituitary gland, and that there may be a pathway in which interferons stimulate PRL secretion through interleukin-6 release.

Analysis of Variance↗