Efficient method for the quantitation of urinary leukotriene E4: extraction using an Empore C18 disk cartridge.

We describe here an efficient procedure for the precise quantitation of leukotriene E4 (LTE4) in a small volume of urine, which was achieved mainly by the use of an Empore extraction disk cartridge. After addition of [3H]LTE4 to 2 ml of urine, an Empore C18 cartridge was used for initial extraction of the urine, which resulted in the extraction of LTE4 in a small volume of solvent. The eluate could then be injected onto a high-performance liquid chromatography column without further concentration.

Induction of human IgE synthesis in B cells by a basophilic cell line, KU812.

Induction of human IgE synthesis in B cells requires, in addition to IL-4 or IL-13, a second signal provided by CD40 ligand (CD40L) on activated Th2-type CD4+ T cells that do not or weakly express Fas ligand (FasL). Mast cells and basophils also produce IL-4 or IL-13 and express CD40L after immunologic or pharmacologic stimulation, although it is unknown whether these cells express FasL. This study investigated the capacity of KU812 cells, a human basophilic cell line, to produce IL-4 and IL-13, to express CD40L and FasL, and to induce IgE and IgG4 synthesis in human normal B cells.

Possible role of nuclear factor-kappa B activity in germline C epsilon transcription in a human Burkitt lymphoma B cell line.

Nuclear factor-kappa B (NF-kappa B) plays a broad role in gene regulation, but it is not evident whether NF-kappa B acts as a messenger system for germline C epsilon transcription. We report here that the signaling cascade triggered by interleukin-4 (IL-4) or anti-CD40 monoclonal antibody (mAb) participates in NF-kappa B activation responsible for germline C epsilon transcription in a human Burkitt lymphoma B cell line, DND39. Both IL-4 and anti-CD40 mAb induced activation of phosphatidylinositol 3-kinase (PI3-kinase), translocation of a zeta isoform of protein kinase C, and nuclear expression of NF-kappa B.

[Environmental allergens and allergic diseases].

Prevalence of allergic diseases in Japan has been increasing these 30 years. About 3%/5% of adult/ child general population are suffering from bronchial asthma these days. Housedust-mite, pollen, fungi and food have been identified as causative allergen.

Allergenicity of acid protease secreted by Candida albicans.

We have previously reported the cases of Candida albicans (C. alb) acid protease (CAAP)-induced atopic asthma. In this study, the allergenicity of the released enzyme CAAP was examined among asthmatic patients with positive immediate skin response to crude C.

Involvement of nuclear factor-kappa B activation in IgE synthesis in human B cells.

Nuclear factor-kappa B (NF-kappa B) is a transcription factor that binds to the consensus DNA sequence in the cis-acting elements of various genes. Although NF-kappa B activates the expression of many genes involved in immune and inflammatory responses, little is known about the role of NF-kappa B activation in the induction of IgE synthesis in human B cells. Therefore we first examined the participation of NF-kappa B in germline C epsilon transcription in a human Burkitt lymphoma B cell line, DND39.

[Thromboxane A2 receptor antagonist in asthma therapy].

Lung tissues produce a large amount of Thromboxane (Tx) A2. In addition to platelet aggregation and artery smooth muscle contraction, TxA2 strongly induces airway smooth muscle contraction and bronchial hyperresponsiveness. Not only TxA2, but many arachidonate cyclooxygenase metabolites such as PGD2, PGF2 alpha, PGH2, and others stimulate TP (PGH2/TxA2) receptor and can take a pathophysiological role for bronchial asthma.

[Bronchial asthma as an inflammation and the use of inhaled steroid].

Recently, bronchial asthma has been widely recognized as a chronic inflammatory disorder of the airways. In the course of inflammation, many kinds of inflammatory cells such as lymphocytes, especially Th2 lymphocytes, mast cells, eosinophills, bronchial epithelial cells interact each other by so-called cytokine network. Bronchial hyperreactivity, which is a characteristic phenomenon in bronchial asthma, is induced as a result of bronchial inflammation.

Histamine augments VCAM-1 expression on IL-4- and TNF-alpha-stimulated human umbilical vein endothelial cells.

Interaction between vascular cell adhesion molecule-1 (VCAM-1) on endothelial cells and CD49d molecules on eosinophils is thought to mediate the selective infiltration of eosinophils into inflamed tissues in allergic disease. IL-4 and TNF-alpha are associated with allergic conditions, and they have been shown to selectively augment VCAM-1 expression on endothelial cells, suggesting that they may be responsible for VCAM-1 expression in allergic disease. We used immunocytochemical staining analysis to examine the effect of chemical mediators, including histamine, leukotrienes and platelet-activating factor (PAF), on VCAM-1 expression in IL-4- and TNF-alpha-stimulated endothelial cells.

Evidence for a role of phosphatidylinositol 3-kinase in IL-4-induced germline C epsilon transcription.

Association of interleukin-4 receptor (IL-4R) with phosphatidylinositol 3-kinase (PI3-kinase) has been demonstrated as the proximal event of IL-4 signaling. We investigated the role of this enzyme in the IL-4 signaling pathway in a human Burkitt lymphoma B cell line, DND39, that expresses germline C epsilon transcripts in response to IL-4. Stimulation of DND39 cells with IL-4 resulted in an accumulation of PI-3-monophosphate as well as a decrease of PI-4,5-bisphosphate, which were abrogated by wortmannin, a potent inhibitor of PI3-kinase.

[The role of cytokines and cell adhesion molecules in allergic inflammation].

The mechanisms of late phase allergic reaction (LAR) have been investigated by many researchers in the past decade, since LAR was recognized clinically to be more important than early phase allergic reaction (EAR). The selective eosinophil recruitment in LAR can be explained by the hypothesis of the cascade of cytokines and adhesion molecules. Since cytokines were initially identified as the factors derived from activated T cells, T cells were thought to play an important role in the LAR in the late 1980s.

Species-specific measurement of the second group of Dermatophagoides mite allergens, Der p 2 and Der f 2, using a monoclonal antibody-based ELISA.

Background: The group 2 Dermatophagoides mite allergens, Der p 2 and Der f 2, were known to be highly crossreactive, and previous assays to measure Der p 2 and Der f 2 were not species-specific.

Objective: The aim of this study was to develop a monoclonal antibody-based ELISA (MoAb-ELISA) to species-specifically measure Der p 2 and Der f 2.

Methods: The MoAb-ELISA for Der p 2 and Der f 2 was performed using species-specific MoAbs for Der p 2 and Der f 2 and a biotinylated second MoAb which recognized a common epitope on both Der p 2 and Der f 2.

Effect of L-tryptophan products on function of human eosinophils: investigation of the causal mechanisms of eosinophilia myalgia syndrome associated with L-tryptophan products.

Contaminants in the L-tryptophan products, known as peak-E and peak-5, at a concentration of 1-10 micrograms/ml had the ability to elicit chemokinetic migration of eosinophils. Purified eosinophils adhered to peak-E- or peak-5-stimulated human umbilical vein endothelial cells, and this adherence was inhibited by the presence of antibody to intercellular adhesion molecule-1, but not by vascular cell adhesion molecule-1 antibody. Neither contaminant affected the expression of integrins, e.

One case of pemphigus vulgaris observed in the oral cavity.

We report on a case of pemphigus vulgaris with lesions in the oral cavity as well as an outline of differential diagnosis and treatment. Changes in the pemphigus antibodies in blood assisted in judging the therapeutic effects along with the therapeutic course.

IgE-mediated basophil releasability is influenced by intrinsic factors and by IgE on the cell surface.

Experiments were done to clarify the mechanisms associated with releasability of histamine. First, washed leukocytes from 23 asthmatic patients sensitive to mite allergen were challenged with Der p 1, a major allergen isolated from Dermatophagoides pteronyssinus, or anti-IgE. A significant correlation was observed between the ratio of Der p 1-specific IgE titer to total IgE level (S/T) in the patient's plasma and either the reactivity (maximal percentage of histamine release; rs = 0.

Functional significance of IL-4 receptor on B cells in IL-4-induced human IgE production.

IL-4 with the IgE-inducing activity is shown to upregulate the expression of IL-4 receptor (IL-4R) on lymphocytes. Antisense strategy was used that aimed at investigating the significance of IL-4-induced upregulation of IL-4R on B cells in human IgE production. When an antisense phosphorothioate oligodeoxynucleotide to IL-4R (S-oligo 1) was added to B cells together with IL-4, the agent selectively abrogated the upregulation of IL-4R without affecting its constitutive level expression.

[Early effects of intravenous steroid therapy on cell recruitment in sputum of chronic asthmatics].

Steroid is the key drug in the asthma therapy but not well known to play the role on the airway inflammatory cells. We examined sputum cells of acute excerbated 11 chronic adult asthmatics before and 2-3 hours after intravenous steroid and aminophylline therapy. There was no changes in the percent count of living cell, epithelial cell, metachlomatic cell, macrophage and neutrophil before and after treatment.

Inhibition of IL-4 receptor up-regulation on B cells by antisense oligodeoxynucleotide suppresses IL-4-induced human IgE production.

IL-4 is shown to up-regulate its own receptor (IL-4R) on human lymphocytes, but the functional significance of up-regulated IL-4R is not clear regarding IgE production. This study investigated the possible role of IL-4-induced up-regulation of IL-4R on B cells in the induction of human IgE synthesis by means of antisense strategy. Among three antisense oligodeoxynucleotides designed against the downstream of translation initiation site of IL-4R cDNA, S-oligo 1, complementary to nucleotide 1-24, showed the strongest inhibition of the constitutive expression of IL-4R on Daudi cells.

Comparison of anti-allergic activities of the histamine H1 receptor antagonists epinastine, ketotifen and oxatomide in human leukocytes.

The effects of three histamine H1 receptor antagonists, epinastine (CAS 80012-43-7, WAL-801 CL), ketotifen (CAS 34580-13-7) and oxatomide (CAS 60607-34-3), on mediator release have been studied in human peripheral leukocytes. When leukocytes from asthmatic patients sensitive to mite were stimulated with the allergen, epinastine inhibited histamine release with a concentration required for 50% inhibition (IC50) of 3 x 10(-5) mol/l and leukotriene C4 generation. On the other hand, ketotifen or oxatomide showed little inhibiting effect on histamine release elicited with the allergen.

[Review of epidemiological studies on adult bronchial asthma in Japan].

The prevalence of adult asthma in Japan has tripled since the 1960 s (1.2%- > 3.14%).

Estimation of Der p and Der f I quantities in the reference preparations of Dermatophagoides mite extracts.

A monoclonal antibody-based enzyme-linked immunosorbent assay (MoAb-ELISA) was developed to measure the major Dermatophagoides mite allergens, Der p I and Der f I. The assay was highly species-specific and sensitive. Using this assay system, the absolute mass unit of Der p I and Der f I in the reference preparations of the extracts was estimated.

Recombinant soluble form of the human high-affinity immunoglobulin E (IgE) receptor inhibits IgE production through its specific binding to IgE-bearing B cells.

A recombinant soluble form of the alpha subunit of the human high-affinity receptor for IgE (rsFc epsilon RI alpha), one of the potent IgE-binding molecules, was tested for its ability to regulate IL-4-induced IgE synthesis by human lymphocytes. Addition of rsFc epsilon RI alpha to cultures induced a dose-dependent inhibition of the T cell-dependent and independent synthesis of IgE. The suppression of IgE synthesis was observed at the protein and the mRNA levels, and it was IgE class specific.

Allergen-specific human IgE helper T cell lines derived from patients allergic to Japanese cedar pollen.

To study the regulatory mechanism of allergen-dependent human IgE synthesis, Cry j I-specific and interleukin 4 (IL-4)-producing CD4+ T cell lines (SN-4 and SS-12) were established from 2 patients allergic to Japanese cedar pollen who highly expressed IL-4 mRNA in T cells in response to Cry j I stimulation. Upon stimulation of SN-4 and SS-12 cells with Cry j I, IL-4 production, which was observed at the protein and the mRNA levels, was induced in an HLA-DR-restricted manner, using autologous and allogeneic antigen-presenting cells. In addition to IL-4, not only considerable amounts of IL-5 and IL-6 but also very small amounts of IL-2 and interferon-gamma (IFN-gamma) were secreted by SN-4 and SS-12 cells, indicating that they fit into the Th2-like phenotype.

[Clinical significance of cat allergens in adult asthmatics--a study of inhalation provocation tests].

We performed a bronchial provocation tests on 40 adult cat sensitive asthmatics to study the clinical significance of cat allergens. Immediate asthmatic responses (IAR) and/or late asthmatic responses (LAR) were provoked in 29 out of the 40 cases. IAR alone was provoked in 12 cases (30%), LAR alone in 7 cases (17.

[A study of hyperreactivity test thresholds over the last 9 years in asthmatics].

We have regularly performed acetylcholine hyperreactivity tests (Ach-HRT) on asthmatics on their first visit to our allergy clinic, and gained the impression that Ach-HRT thresholds have risen over the last nine years. We studied the thresholds in four different years: 1983, '85, '87 and '91 in 578 male cases and 680 female cases. Our findings were as follows.