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Biomedical subjects

N Kanbe

Publications and source records attributed to N Kanbe.

At least 19 recordsLinked to original sources
PubMed2000

Nerve growth factor prevents apoptosis of cord blood-derived human cultured mast cells synergistically with stem cell factor.

BACKGROUND: Stem cell factor (SCF) has been identified as a critical survival factor of human mast cells. Other cytokines which possess survival promotion activity on human mast cells are less known. OBJECTIVE: We examined the survival promotion activity of nerve growth factor (NGF) on cord blood-derived human cultured mast cells. METHODS: Expression and function of NGF receptors on the mast cells were examined by RT PCR, flowcytometric analysis, immunoprecipitaion and western blotting. The survival promotion activity of NGF to the mast cells was examined. To evaluate the proliferating activity of NGF on the human cultured mast cells, flow cytometric analysis with propidium iodide staining was applied. To confirm whether the human mast cell growth activity of NGF was caused by a suppression of apoptosis, the proportion of the cells containing in situ DNA fragmentation was counted. RESULTS: The human cultured mast cells expressed the high affinity receptor p140trk but not the low affinity receptor p75LNGFR. NGF induced the phosphorylation of p140trk. NGF alone could not support the survival of the mast cells, however, the addition of NGF to the culture medium containing recombinant SCF led to a significant increase of the number of survival mast cells. No significant changes of the cell cycle from G0/G1 phase to the S/G2 + M phases were observed by NGF. In contrast, the addition of NGF to the medium with SCF showed a significant inhibitory effect on the apoptosis of the mast cells. CONCLUSION: NGF may act as a key factor to promote the survival of human mast cells synergistically with SCF through the prevention of apoptosis.

Apoptosis↗
PubMed2000

Production of fibrogenic cytokines by cord blood-derived cultured human mast cells.

BACKGROUND: Mast cells are a potential source of cytokines, but their contribution to nonallergic inflammatory conditions, such as fibrosis, remains unclear. OBJECTIVE: We investigated whether cord blood-derived cultured human mast cells could produce fibrogenic cytokines by IgE-mediated activation. METHODS: Mast cells were obtained from human cord blood mononuclear cells by culture with stem cell factor and IL-6. The mast cells were incubated with human myeloma IgE and were activated with anti-IgE. The expression of messenger RNA for fibrogenic cytokines was examined by the reverse-transcription polymerase chain reaction, and cytokine protein was assayed by enzyme-linked immunosorbent assay, or bioassay. RESULTS: Cultured human mast cells constitutively expressed mRNA for transforming growth factor-beta(1), and its expression was not increased by anti-IgE stimulation. The cells released this factor into the culture medium spontaneously, which showed bioactivity after heat treatment. The mast cells also expressed mRNA for platelet-derived growth factor A, which was enhanced with a peak at 3 hours by stimulation with anti-IgE. Conditioned medium from nonactivated mast cells did not contain basic fibroblast growth factor, but this cytokine was released into the medium in a time-dependent manner after stimulation with anti-IgE. CONCLUSION: Human mast cells activated by IgE-mediated stimulation show production of fibrogenic cytokines that varies depending on the cytokine, which suggests possible involvement of mast cell cytokines in the development of fibrosis.

Cells, Cultured↗
PubMed1999

Human mast cells produce matrix metalloproteinase 9.

Extracellular matrix-destructive enzymes, like matrix metalloproteinases (MMP), have been recognized in the process of inflammation and tissue remodeling and repair. The affected tissues often contain markedly increased numbers of mast cells. Although mast cells are capable of activating latent collagenase and proMMP, it has so far been unknown whether human mast cells themselves produce and secrete MMP9. In this study, MMP9 production by cord blood-derived cultured human mast cells and HMC-1 human mast cells was examined by reverse-transcriptase PCR, gelatin zymography and Western blot analysis using an antibody against MMP9. Cultured mast cells and HMC-1 cells treated with phorbol 12-myristate 13-acetate were shown to express MMP9 mRNA, and the cultured conditioned media from these cells showed gelatinolytic activity, identical with MMP9. Immunohistochemical examination was performed to detect MMP9 in tissue mast cells; mast cells localized in the skin, lung and synovial tissue showed strongly positive reactions for MMP9. Thus, these findings indicate that human mast cells can produce MMP9, which might contribute to extracellular matrix degradation and absorption in the process of allergic and nonallergic responses.

Base Sequence↗
PubMed1999

Response to cyclosporin and low-dose methylprednisolone in aggressive systemic mastocytosis.

BACKGROUND: There is no effective treatment for aggressive systemic mastocytosis. OBJECTIVE: The purpose of this study was to investigate the effect of cyclosporin and low-dose methylprednisolone in a 64-year-old man with aggressive systemic mastocytosis. METHODS: Immunohistochemical studies were done on biopsy specimens from the skin and other organs. Mast cells, predominantly containing tryptase, were derived from human umbilical cord blood cells cultured in the presence of stem-cell factor and IL-6. IgE-sensitized cultured human mast cells were activated by anti-IgE, and the effect of cyclosporin on histamine release was investigated. In addition, blood and urine levels of various mediators were measured in the patient before and after therapy. RESULTS: Biopsy specimens of the patient's skin lesions showed an increase of mast cells; cells containing tryptase (but not chymase) comprised 20% to 50% of the skin mast cells. Histamine release from activated cultured mast cells was inhibited by cyclosporin in a concentration-dependent manner. When the patient was treated with cyclosporin and low-dose methylprednisolone, he showed a good response. CONCLUSION: Cyclosporin combined with low-dose methylprednisolone may be a reasonable therapy for aggressive systemic mastocytosis.

Bone Marrow Cells↗
PubMed1999

Nerve growth factor release with mast-cell-derived mediators in a patient with systemic mastocytosis after middle-wave ultraviolet irradiation.

BACKGROUND: The symptoms of a 56-year-old man with systemic mastocytosis became worse with exposure to sunlight. We evaluated mast-cell-derived mediators and cytokines before and after exposure to ultraviolet light in the patient. METHODS: The patient was irradiated with middle-wave ultraviolet light, so-called ultraviolet light B, and the levels of mediators and cytokines were measured serially. The point mutation Asp816Val in c-kit was investigated by analyzing polymerase chain reaction products from the complementary DNA of peripheral blood mononuclear cells. RESULTS: Before irradiation, the levels of mast-cell-derived mediators and metabolites were elevated. Among various cytokines measured, including soluble c-kit and stem cell factor, only the level of nerve growth factor was elevated. After irradiation, the nerve growth factor level was further increased along with the levels of mast-cell-derived mediators and metabolites. The point mutation Asp816Val in c-kit was not detected in peripheral blood mononuclear cells. CONCLUSIONS: Middle-wave ultraviolet light may activate mast cells to release nerve growth factor and mediators in systemic mastocytosis.

Cytokines↗
PubMed1999

Cord blood-derived human cultured mast cells produce transforming growth factor beta1.

BACKGROUND: Mast cells frequently accumulate at the site of fibrosis and their contribution has been suspected in the pathogenesis of fibrotic conditions. However, it still remains unknown whether human mast cells synthesize transforming growth factor beta1 (TGF-beta1). OBJECTIVE: We have investigated whether cord blood-derived human cultured mast cells express messenger RNA (mRNA) for TGF-beta and produce bioactive TGF-beta1. METHODS: Mast cells were obtained by culturing mononuclear cells from cord blood in the presence of stem cell factor and interleukin-6. Expression of mRNA for TGF-beta1 was examined by the method of reverse transcriptase-polymerase chain reaction (RT-PCR). Immunocytochemical staining for TGF-beta and growth-inhibitory assay using Mv1Lu cells were also performed. RESULTS: The cultured human mast cells constitutively expressed mRNA for TGF-beta1. With calcium ionophore A23187, the intensity of the PCR-amplified band for TGF-beta1 was not increased. Immunocytochemical staining showed that the cultured mast cells were positive for both latency-associated peptide and activated forms of TGF-beta. Bioassay with Mv1Lu cells and R 4-2 mutant cells showed that mast-cell conditioned medium had a bioactivity of TGF-beta1. CONCLUSION: Cord blood-derived human cultured mast cells constitutively express mRNA for TGF-beta1 and produce functional TGF-beta1. Because TGF-beta1 has been shown to be highly fibrogenic, these results may highlight a novel role for human mast cells in tissue fibrosis.

Animals↗
PubMed1999

Cord-blood-derived human cultured mast cells produce interleukin 13 in the presence of stem cell factor.

BACKGROUND: Mast cells have been regarded as a potential source of cytokines. Although the human mast cell line HMC-1 and human lung mast cells have been shown to produce interleukin (IL) 13, it still remains uncertain whether cord-blood-derived human cultured mast cells produce IL-13. METHODS: Human cultured mast cells were raised from cord blood cells in the presence of stem cell factor (SCF) and IL-6. Levels of IL-13 mRNA were examined by a semiquantitative reverse transcriptase-polymerase chain reaction. IL-13 levels in the supernatants were measured with an enzyme-linked immunosorbent assay. RESULTS: When the IgE-sensitized cultured mast cells were activated with anti-IgE, mRNA for IL-13 was amplified with a peak at 3 h after the stimulation. IL-13 was not detected in the supernatants of the activated mast cells in the absence of SCF, whereas the mast cells secreted significant amounts of IL-13 after the stimulation in the presence of SCF. Calcium ionophore A23187 also stimulated the mast cells to release IL-13 into the supernatant in the presence of SCF. CONCLUSIONS: These observations suggest that human mast cells can produce IL-13 under the condition with SCF. The cord-blood-derived human cultured mast cells will help in studying the functional properties of human mast cells in allergic diseases.

Calcimycin↗
PubMed1998

Cloning and tissue distribution of a novel serine protease esp-1 from human eosinophils.

We have cloned a novel serine protease designated as esp-1 from human eosinophils. The amino acid sequence deduced from the cDNA showed that ESP-1 comprises a signal peptide of 18 amino acids, a propeptide of 23 amino acids, an active form sequence of 273 amino acids starting from an Ile-Val-Gly-Gly-Glu motif, the catalytic triad of serine proteases that has been characterized as the essential amino acid residues for the proteolytic activity, and a hydrophobic amino acid stretch in the carboxyl terminus, suggesting this enzyme is a novel membrane-type serine protease. The tissue distributions of esp-1 expression revealed that this protease is not only expressed in human eosinophils, but also widely expressed in mononuclear cells and various tissues other than skeletal muscle and kidney and is most abundant in testis and prostate, and moderately so in lung, spleen and pancreas.

Amino Acid Sequence↗
PubMed1998

Phase-contrast microscopic studies using cinematographic techniques and scanning electron microscopy on IgE-mediated degranulation of cultured human mast cells.

BACKGROUND: Isolating human mast cells is a laborious procedure. Recently, cultured human mast cells raised from umbilical cord blood cells have become available. It is necessary to investigate whether IgE-mediated activation of these cells is mediated by exocytosis. OBJECTIVE: To verify IgE-mediated activation of these cultured human mast cells morphologically. METHODS: The mast cells were raised from human umbilical cord blood cells in the presence of stem cell factor and interleukin-6. IgE-sensitized cultured human mast cells were activated by anti-IgE, and morphological changes of the cells were examined under phase-contrast microscopy using cinematographic techniques and scanning electron microscopy. Histamine release from the cells was measured with high-performance liquid chromatography. RESULTS: Under the condition in which a significant histamine release was observed from the mast cells, phase-contrast microscopy showed that the cultured human mast cells became swollen and extruded granules. Scanning electron microscopy disclosed the extrusion of smooth and round bodies from pores formed on the activated mast cell surface. CONCLUSION: IgE-mediated histamine release from cultured human mast cells is accompanied by exocytosis morphologically, indicating that cultured human mast cells will help in studying the functional properties of human mast cells.

Cell Degranulation↗
PubMed1998

Carnoy's fixative reduces the number of chymase-positive cells in immunocytochemical staining of cord-blood-derived human cultured mast cells.

KleinJan et al. (Allergy 1996;51:614-20) reported that Carnoy's fixative reduced the number of chymase-positive mast cells in the nasal mucosa. Therefore, in the present study, we investigated whether Carnoy's fixative reduces the number of chymase-positive cells from cord-blood-derived human cultured mast cells when compared with other types of fixatives. Human mast cells were obtained by culturing cord-blood-derived CD34-positive cells in the presence of stem cell factor and interleukin-6. Staining procedures of the cells in fixation with Carnoy's fixative and with other fixatives gave no differences among the number of tryptase-positive cells, whereas fixation with Carnoy's fixative for 15 min gave a significant decrease in the number of chymase-positive cells compared with acetone for 10 min. The number of chymase-positive cells decreased in a time-dependent manner under fixation with Carnoy's fixative, indicating that Carnoy's fixative had a negative effect on the number of chymase-positive cells from cord-blood-derived human cultured mast cells.

Acetic Acid↗
PubMed1997

Heterogeneity of mast cells in mastocytosis and inhibitory effect of ketotifen and ranitidine on indolent systemic mastocytosis.

BACKGROUND: Mastocytosis is a disorder of mast cell proliferation that occurs in both cutaneous and systemic forms. The most frequent site is the skin. OBJECTIVE: The mast cell subtype of two patients with mastocytosis was investigated. METHODS: Immunohistochemical studies were performed on the skin or gastric mucosa or both of the two patients. Blood and urine levels of various mediators were measured for one patient. RESULTS: Mast cells containing tryptase and chymase were the only type seen in the skin lesions of an 11-month-old boy with urticaria pigmentosa. Mast cells containing tryptase were predominant in lesions of the skin and gastric mucosa of a 41-year-old man with indolent systemic mastocytosis. However, mast cells containing tryptase and chymase were predominant in the nonlesional and the normal skin of this patient. Tryptase-positive cells were more numerous in lesional skin than nonlesional skin and normal skin. Elevated blood and urine levels of various mediators were decreased by means of combination therapy with ketotifen and ranitidine. CONCLUSION: In indolent systemic mastocytosis, mast cell dynamics involve only cells containing tryptase. Release of mediators from mast cells may be inhibited by means of combination therapy with histamine H1 and H2 receptor antagonists.

Adult↗
PubMed1997

65-kilodalton protein phosphorylation in human peripheral blood eosinophils.

To evaluate the presence of protein phosphorylation in peripheral blood eosinophils, venous blood was drawn from normal healthy volunteers. Eosinophils were isolated on a Percoll gradient and were incubated with [gamma32P]ATP in the presence of Mg2+. After stopping the reaction, SDS-PAGE was performed and autoradiographs were prepared to determine the incorporation of 32P into proteins. Eosinophils developed at least 24 protein bands below 116.25 kD by SDS-PAGE. In the autoradiographs, one distinct radioactive band was observed with a molecular weight of 65 kD. 32P incorporation into the 65-kD band was dependent on Mg2+ concentration and maximal response was observed at concentrations of 2-6 mM MgCl2. 32P incorporation into the band was dependent on the reaction time and temperature of the reaction system. Acid hydrolysis showed that [32P]phosphate radioactivity in the cells was present primarily as phosphoserine, indicating the presence of 65-kD protein phosphorylation in human peripheral blood eosinophils.

Adult↗
PubMed1988

[Gastrointestinal carcinoma with skin diseases from the standpoint of surgery].

We encountered 21 patients with gastrointestinal carcinoma accompanied with skin diseases such as acanthosis nigricans, dermatomyositis, pemphigoid, erythrodermia, ichthyosis vulgaris, herpes zoster, Leser-Trélat's sign, Recklinghausen's disease and Gardner's syndrome. We have discussed such cases from the standpoint of surgery. There were 16 cases (76%) with gastric carcinoma, which were almost in the far advanced stage. Carcinomas of the other patients were seen in the esophagus, liver, pancreas, small intestine and rectum. Although skin lesions appeared before the onset of gastrointestinal symptoms in almost all cases, detection of carcinomas was very difficult until the symptoms emerged. In 6 of 21 patients (29%), carcinomas were detected by only skin manifestation, and they proved to be gastric cancer in all cases. Although the relationship of gastrointestinal carcinoma and skin diseases was unclear, we presented a 70-year-old woman with acanthosis nigricans accompanied with gastric cancer, whose skin manifestation was reduced by the therapy for gastric cancer. Therefore, some of these skin manifestation may be a symptom of the biological characteristics of the gastrointestinal carcinoma.

Acanthosis Nigricans↗
PubMed1986

A case of primary biliary cirrhosis associated with generalized morphea.

A 50-year-old female with primary biliary cirrhosis associated with generalized morphea is reported. She had suffered from multiple painful indurated plaques on the trunk which was diagnosed as generalized morphea; subsequently, primary biliary cirrhosis was diagnosed on the basis of her hepatic dysfunction. Although progressive systemic sclerosis has been reported to occur in patients with primary biliary cirrhosis, this localized form of cutaneous sclerosis has not previously been reported. Since generalized morphea is one of the most common manifestations of chronic graft-versus-host disease, and primary biliary cirrhosis is known to have a hepatic histology resembling chronic graft-versus-host disease, this case report may represent evidence in support of a common pathogenesis for the two entities.

Female↗
PubMed1981

The renal and hemodynamic effects of furosemide in acute myocardial infarction.

Renal and hemodynamic effects of furosemide were studied in 30 patients with acute myocardial infarction. When marked congestion of lungs was absent, the effects of furosemide were somewhat different from those of recent reports. The changes of pulmonary arterial pressure and wedge pressure were biphasic; there was first pressure rise in many occasions followed by reduced pressure; the latter is commonly believed to be the beneficial effect of furosemide. In these patients without marked congestion but in an acute state of myocardial infarction, within 5 min of furosemide administration, there was marked increase in urine volume, sodium and potassium excretion along with increases in creatinine clearance which is commonly found in the other conditions, but is in contrast to the recent reports on patients with acute myocardial complicated with pulmonary congestion.

Aged↗