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Biomedical subjects

Michihiro Hide

Publications and source records attributed to Michihiro Hide.

At least 19 recordsLinked to original sources

Bone marrow derived mast cell acquire responsiveness to substance P with Ca(2+) signals and release of leukotriene B(4) via mitogen-activated protein kinase.

Substance P (SP) selectively activates mast cells that reside in connective tissues. We studied the reactions of bone marrow-derived mast cells (BMMC) of three mouse strains, cultured with or without fibroblasts. BMMC co-cultured with fibroblasts, but not those cultured alone, increased intracellular Ca(2+), released LTB(4) and histamine in response to SP. PD098059 significantly inhibited the release of LTB(4), but not histamine in all strains. SB203580 failed to reduce or slightly impaired the release of LTB(4). These results suggest that mast cells undergo maturation under the influence of fibroblasts, acquiring the responsiveness to SP with Ca(2+) signals and predominantly ERK-MAP kinase.

Animals↗

Fucoidan prevents C epsilon germline transcription and NFkappaB p52 translocation for IgE production in B cells.

Fucoidan, a dietary fiber contained in seaweed, reduces the increase of antigen-specific IgE in mice exposed to ovalbumin. In this study, we investigated the effect of fucoidan on IgE production and intracellular events in B cells in vitro. Fucoidan inhibited the production of IgE and C epsilon germline transcription in murine B cells induced by IL-4 (100 ng/ml) and anti-CD40 antibodies (10 microg/ml), whereas it stimulated cell proliferation. A significant effect of fucoidan on IgE production was observed when B cells were stimulated with a higher dose (5 microg/ml) of anti-CD40 antibodies, but not when stimulated with lower doses (1.25, 2.5 microg/ml), regardless of the IL-4 concentrations. Moreover, nuclear translocation of NFkappaB p52, but neither that of NFkappaB p65, nor the phosphorylation of JAK1 and STAT6 was reduced by fucoidan. These results suggest that fucoidan inhibited IgE production by preventing the NFkappaB p52-mediated pathways activated by CD40.

Animals↗

The SPR signal in living cells reflects changes other than the area of adhesion and the formation of cell constructions.

Surface plasmon resonance (SPR) sensors detected large angle of resonance (AR) changes, when RBL-2H3 rat mast cells were cultured and activated on a sensor chip. Here, we demonstrated that PAM212 mouse keratinocytes also showed a large change in AR, when EGF-stimulated. We explored these changes due to intracellular reactions, through the relationship between the AR and the area of cell adhesion, using confocal microscopy for RBL-2H3 cells and PAM212 cells. The effect of Mycalolide B and Toxin B, inhibitors for cell motility, on AR was observed using RBL-2H3 cells. Measuring AR in the presence of various numbers of non-stimulated cells demonstrated that AR and cell density were proportional. However, the AR increase in response to antigen was 35% higher than that expected by solely an increase of the cell adhesion area. Moreover, the AR with PAM212 cells decreased following a transient increase in response to EGF, whilst the area of cell adhesion remained at an increased level. Furthermore, the treatment of RBL-2H3 cells with either Mycalolide B or Toxin B slightly inhibited, but never abolished the AR increase induced by antigen. These treatments abolished all morphological changes, including ruffling and the increase of cell adhesion area observed by light microscopy. These results suggest that AR changes reflect intracellular events rather than changes in the size of the area to which cells adhere.

Cell Adhesion↗

The extract of syngeneic keratinocytes enhances IgE production from BALB/c mouse splenic lymphocytes in vitro.

The increase of serum IgE levels is closely associated with atopic dermatitis. We have previously revealed that cellular extract of PAM212 cells (PAM-extract), BALB/c mouse keratinocyte cell line, induced a remarkable increase of serum IgE levels, in vivo, when subcutaneously injected into BALB/c mice. However, precise mechanism of IgE-increasing activity was unclear. To elucidate the mechanism of IgE-increase in sera of BALB/c mice induced by PAM-extract, we explored the direct influence of PAM-extract on immunoglobulin production and class-switching in the culture of splenic lymphocytes and purified B-cells, in vitro. Splenic lymphocytes or purified B-cells obtained from BALB/c mice were cultured with various combinations of IL-4, anti-CD40 antibody, and PAM-extract for 7 days. IgE and IgG concentrations of culture supernatants were measured by ELISA. Epsilon germ-line transcriptions were assessed by RT-PCR from the cultured cells. IgE and IgG concentrations in culture supernatant of splenic lymphocytes were increased by an addition of PAM-extract in the presence of both IL-4 and anti-CD40 antibody. Epsilon germ-line transcript was also induced in parallel to the increase of IgE production. Similar results were obtained when purified B-cells were employed in stead of whole splenic lymphocytes. The enhancement of IgE production in vitro was also observed, when splenic lymphocytes of CBAj mouse were cultured with cellular extract of KCMH-1 cells, CBAj mouse keratinocyte cell line. The cellular extract of keratinocyte promotes immunoglobulin class-switching to IgE and IgE production from mouse splenic B-cells in an IL-4- and CD40-stimuli-dependent manner. Such enhancement may account for the increase of serum IgE in patients with dermatitis in association with a Th2 microenvironment.

Animals↗

Anaphylactic shock caused by exposure to sea anemones.

BACKGROUND: Since the first report of a dog that developed severe systemic symptoms in response to a second injection of sea anemone toxin by Richet and Portier in 1902, no clear human cases of anaphylaxis related to exposure to sea anemones has been reported in the literature. METHODS: A 24-year-old man with an episode of local urticaria on his first contact with a sea anemone (Stichodactyla haddoni), developed dyspnea, severe urticaria and hypotension on exposure to water containing the dead bodies of the organism. To study whether this reaction was mediated by antigen-specific IgE, we performed a histamine release test with blood, Western blotting with serum and lymphocyte proliferating test with peripheral blood mononuclear cells of the patient, for the homogenate of sea anemones. RESULTS: The homogenate of sea anemones induced histamine release from the blood of the patient, but it also induced histamine release from the blood of control subjects. Moreover, it also caused hemolysis of blood of all donors. However, Western-blotting demonstrated the presence of an 86 kd protein-specific IgE in the serum of the patient. CONCLUSIONS: Protein antigen(s) in sea anemones may cause anaphylactic shock under the influence of the cytolytic effects and/or lymphocyte-stimulating activity elicited by the toxin of sea anemones.

Adult↗

[Analysis of the prevalence of subtypes of urticaria and angioedema].

BACKGROUND: The prevalence of various subtypes of urticaria in Japan has not been analyzed. METHODS: We have classified 260 (105 males and 155 females) out patients with urticaria based on the classification in Guidelines for the diagnosis and treatment of urticaria and angioedema" prepared by the Japanese Dermatological Association with slight modification. RESULTS: The urticaria in 200 out of 260 (76.9%) patients was idiopathic, and 101 (38.8%) patients showed more than one type of urticaria or angioedema. Among 146 patients with chronic urticaria, 90 (61.6%) were complicated by other types of urticaria; 59 (40.4%) by factitia, 29 (19.9%) by angioedema, and 12 (8.2%) by other types of urticaria. Deep pressure urticaria has been assumed as a rare type of urticaria in Japan, but was diagnosed in 5 patients with other types of urticaria. CONCLUSION: Most of the subtypes of urticaria could be diagnosed mainly by history and physical examinations. Correct diagnosis of the subtypes of urticaria should be the basis of better understanding and the treatment of this disease.

Angioedema↗

[Two cases of food-dependent exercise-induced anaphylaxis difficult to evoke symptoms by provocation test].

We report two cases of food-dependent exercise-induced anaphylaxis (FDEIA), which were hardly induced by provocation test in the hospital. Case 1: A 28-years-old Japanese female suffers repeated episodes of sternutation, nasal discharge and edema of eyelids after wheat ingestion of wheat followed by exercise. Case 2: A 14-years-old Japanese male suffers repeated episodes of wheal formation on whole body and dyspnea after lunch containing apple followed by exercise. Both of them had never developed symptoms by either ingestion or exercise alone. Provocation tests were performed on admission by combinations of the ingestion of suspected foods, exercise, and aspirin, but no symptoms were reproduced by any combination of them. After discharge, case 1 reproduced symptoms during exercise after the ingestion of wheat under prostration and cold climate. Case 2 reproduced symptoms during exercise after ingestion of apple when he suffered from common cold. Warm and comfortable condition in admission may make it harder to evoke symptoms by the provocation test. Frigidity, cold, prostration, and stress should be reckoned with in the provocation test to improve the accuracy of diagnosis for FDEIA.

Adolescent↗

IL-4 modulates the histamine content of mast cells in a mast cell/fibroblast co-culture through a Stat6 signaling pathway in fibroblasts.

IL-4 plays a crucial role in the pathogenesis of allergic diseases, such as the induction of IgE synthesis and the development of mast cells. To further understand the effect of IL-4 on mast cells in skin, we utilized a mast cell/fibroblast co-culture system as an in vitro model of dermal mast cells. IL-4 induced mast cell growth in the culture with fibroblasts. Immunoblot analysis revealed that IL-4 activated Stat6 in both mast cells and fibroblasts. The over-expression of dominant-negative Stat6 in fibroblasts in the presence of IL-4 decreased the histamine content per mast cell, but not the number of mast cells. In contrast, the over-expression of constitutively-active Stat6 in fibroblasts increased the histamine content per mast cell, indicating that the activation of Stat6 in fibroblasts supports the maturation of mast cells co-cultured with fibroblasts.

3T3 Cells↗

The effectiveness of montelukast for the treatment of anti-histamine-resistant chronic urticaria.

Many patients with chronic idiopathic urticaria are not sufficiently controlled with histamine H(1)-receptor antagonists. Leukotriene receptor antagonists have been reported to be effective for certain cases of urticaria, although their proper application remains to be established. To study the effectiveness of montelukast, a leukotriene receptor antagonist, for the treatment of chronic urticaria that was not controlled by histamine H(1)-receptor antagonists. Twenty-five patients with chronic idiopathic urticaria were treated with 10 mg of montelukast for one week or more, without changing any precedent treatment that they were using before the study including histamine H(1)-receptor antagonists. The effectiveness of montelukast for each patient was evaluated and compared with clinical features and/or backgrounds of the patients. Twelve patients, including six who had been treated with corticosteroids, were evaluated as "markedly improved" or "improved" following treatment with montelukast. There was no statistically significant relation of the effectiveness to the complications with non-steroidal anti-inflammatory drugs (NSAIDs) intolerance, mechanical urticaria, or reactions to autologous serum skin test. However, the patients for whom montelukast was effective were younger (33.2+/-16.3 years, mean +/- SD)(P<0.05, Mann-Whitney test) and their duration of illness shorter (15.9+/-18.3 months) (P<0.005, Mann-Whitney test) than those of patients for whom montelukast was ineffective (45.9+/-15.0 years, 89.6+/-71.7 months). Montelukast may be worth trying for patients with chronic idiopathic urticaria, when the condition is not sufficiently controlled with histamine H(1)-receptor antagonists.

Acetates↗

Proteome analysis of gelatin-bound urinary proteins from patients with bladder cancers.

OBJECTIVES: Proteome analysis with protein separation by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and mass spectrometric measurement is utilized for comprehensive identification of proteins in various biological samples. However, this technique has not been widely applied for the analysis of body fluid for clinical assessments, because large amounts of proteins in fluid such as albumin and globulin hinder the detection of small amounts of proteins with high clinical values. The aim of the present study is to develop a method of proteomic analysis for urine of patients with bladder cancers. MATERIALS AND METHODS: Urine samples collected from 40 patients with bladder cancers, 32 healthy volunteers, and 7 old volunteers with benign prostate hypertrophy were treated with gelatin-beads as a group-specific affinity carrier. The gelatin-bound proteins were subjected to the proteome analysis. RESULTS: Various amounts of matrix metalloproteinase-2 (MMP-2), -9 (MMP-9), fibronectin (FN), and their fragments were identified on 2-D PAGE gels from cancer-bearing patients, but not from normal individuals. Importantly, not only total amounts of these proteins, but also patterns of their distributions on the gels were well matched with the extent of invasion diagnosed with histopathological examinations. Thus, the score including such qualitative patterning of these proteins predicts pathological stages of the cancer. CONCLUSION: The proteomic analysis with gelatin-affinity purification of urine samples is useful not only for the diagnosis of bladder cancers, but also for estimating the extent of tumor invasion.

Biomarkers, Tumor↗

Pelodera strongyloides infestation presenting as pruritic dermatitis.

Pelodera strongyloides is a free-living soil nematode of the order Rhabditida. We report an 18-year-old man with P strongyloides skin infestation. In this case, pruritic follicular papulopustules developed on the buttocks, then the right flank. Skin scrapings revealed many live rhabditiform larvae that were cultured adult worms and hatched ova, identified as P strongyloides . The eruption was treated effectively with topical 1% gamma-hexachlorocyclohexane ointment.

Adolescent↗

Increase in serum IgE levels following injection of syngeneic keratinocyte extracts in BALB/c mice.

Increased serum IgE levels are found in atopic diseases, especially atopic dermatitis. It is generally accepted that the high serum IgE levels are increased by exposure to exogenous antigens. However, exposure to antigens in the absence of proper adjuvants does not usually increase serum IgE. Here, we studied the involvement of skin factor(s) in increasing serum IgE. The soluble fractions were obtained from two keratinocyte cell lines, PAM 212 and KCMH-1, derived from BALB/c and CBA/J mice, respectively. These were injected subcutaneously into parental mice strains at 2-week intervals. The levels of serum immunoglobulins and the release of IgE in vitro from spleen cells were measured by ELISA. The biological activity of the increased serum IgE was assessed by sensitization of RBL-2H3 mast cells with serum. Six injections of PAM 212-derived fraction (PAM extract) consistently increased the serum levels of IgE, IgG1 and IgG2b, but not those of IgM, IgG2a, IgG3 or IgA in BALB/c mice. The high serum levels of IgE were sustained for at least 10 weeks after one or two injections of PAM extract. When RBL-2H3 cells were sensitized with the serum from mice injected with PAM extract, they released histamine in response to anti-IgE. Moreover, spleen cells of these mice produced significantly higher amounts of IgE than those of uninjected mice in the presence of IL-4 and anti-CD40 antibody. On the other hand, no significant increase in immunoglobulins was observed following injection of the soluble fraction from KCMH-1 cells in CBA/J mice. These results suggest that the release of the soluble fraction of keratinocytes and its exposure to immunologically competent cells may induce an increase in serum IgE levels by a Th2-dominant mechanism.

Adjuvants, Immunologic↗

Real-time analysis of ligand-induced cell surface and intracellular reactions of living mast cells using a surface plasmon resonance-based biosensor.

Surface plasmon resonance (SPR)-based sensors have been used to detect the binding between interactive molecules. We applied the SPR technology to the analysis of interactions between living cells and molecules reactive to the cells, using mast cells and mast cell-reactive antigens. The exposure of dinitrophenol-human serum albumin (DNP-HSA), an antigen that stimulates mast cells, to IgE-sensitized mast cells induced a robust and long-lasting SPR signal in a dose-dependent manner. The maximal increase in SPR signal induced by 100 ng/ml DNP-HSA was 0.200 +/- 0.120 angle (mean +/- SD, n = 37), about 1000 times larger than the theoretically expected increase for the simple binding of DNP-HSA to Fc(epsilon)RI, the high-affinity IgE receptor. A small, but similarly prolonged signal was observed when the cells were stimulated by an agonist of the adenosine A3 receptor. The signal induced by DNP-HSA was abolished by genistein, and partially inhibited by phorbol 12-myristate 13-acetate and wortmannin. Interestingly, the signal induced by DNP-HSA was only weakly inhibited by DNP-lysine, suggesting that DNP-lysine manifests its action not by inhibiting, but by modulating the crosslinking of Fc(epsilon)RI. We concluded that SPR sensors can detect biologically significant signals in a real-time manner from the interactions between cells and molecules reactive to the cells.

Androstadienes↗