Recently, the detection of anti-neutrophil cytoplasmic antibodies (ANCA) has gained worldwide acceptance as an important, additional diagnostic tool of Wegener's granulomatosis (WG). A possible pathogenic role of ANCA has been suggested by the finding that the addition of purified IgG-ANCA to healthy neutrophils in vitro causes activation of these cells. But, investigation on neutrophil functions of patients with WG has been rare. To clarify this point, peripheral neutrophil functions (adherence, chemiluminescence and chemotaxis) of patients with active WG were examined. Adherence, chemiluminescence and chemotaxis of patients with WG were significantly suppressed when compared with control subjects. The effects of ANCA positive serum on neutrophil chemiluminescence were also examined. ANCA positive serum did not affect neutrophil chemiluminescence. These results indicate that the mechanism of WG is not simply due to neutrophil activation by ANCA.

View full abstract

Twenty-four steroid-dependent intractable asthmatics were treated with oral prednisolone (PSL) (basic dose) + additional PSL and oral PSL (basic dose) + inhaled beclomethasone (BDP) alternately, to assess the clinical symptoms in each administration period. Equivalent doses of PSL to BDP were calculated. 1. Peak expiratory flow rate (PEF) during additional administration of BDP was markedly higher both in the morning and at night compared with that during additional administration of PSL (p<0.01 and p<0.05 respectively). 2. The relation between additional doses of either PSL or BDP and attack scores was analysed in regression to obtain a dose equivalence line. 3. BDP 400 μg was equivalent to PSL 7.04 mg on average. 4. The equivalent dose of PSL to BDP tended to be lower in cases where the priod of dependency on oral steroids was longer, the basic dose was larger, or the asthma was more severe. These data indicate that the application of inhaled steroids may be useful in the therapeutic management of steroid-dependent intractable asthmatics.

View full abstract

This study was designed to investigate the effect of sustained-release theophylline therapy on serum pyridoxal concentration in children with bronchial asthma. Forty-two children with bronchial asthma were divided into two groups according to duration of theophylline administration: The 22 children in group A had been treated with theophylline for less than 4 weeks, whereas the 20 children in group B had been treated for more than 5 weeks. The results obtained from this study were as follows: 1) The serum pyridoxal concentration in group B was significantly lower than that those in group A (p<0.01). 2) The serum pyridoxal concentration was not significantly correlated with the serum theophylline concentration. These findings suggest that long-term theophylline therapy can depress vitamine B_6 status in children with bronchial asthma. Theophylline-induced seizure may be caused by the possible decreased in gamma-aminobutyric acid concentration in the brain as a result of decrease in gamma-aminobutyric acid concentration in the brain as a result of decrease in vitamin B_6 status, even if the serum theophylline concentrations are within the therapeutic range.

View full abstract

Clinical characterization of patients with allergy to Chamaecyparis obtusa (Cypress) pollen and their specific IgE antibody was investigated by using AlaSTAT assay, a new method for the measurement of antigen-specific IgE antibody. positive rate of antigen-specific antibody in patients with allergic pollinosis was 83.5% for Japanese cedar pollen and 80.0% for Chamaecyparis obtusa pollen. 76.4% of patients were sensitized both to Japanese cedar and Chamaecyparis obtusa pollen, however, 7.9% were sensitized only to the latter. As for clinical symptoms, duration of attack was short and severity was mild or moderate in the patients sensitized only to Chamaecyparis obtusa pollen. Patients who visited hospital in April, in that Chamaecyparis obtusa pollen disseminated in the air predominantly, showed significant elevation in AlaSTAT scores compared with patients who visited in February and March (p<0.05).

View full abstract

Superoxide generation was assessed in highly purified eosinophils (>99.0% pure) with a flow cytometer. Eosinophils from patients with asthma were collected by centrifugation of leukocytes on Percoll density gradients, followed by negative immunoselection with anti-CD16 monoclonal antibodies. For comparison, highly purified neutrophils (<99.5% pure) were collected by a similar method with anti-CD9 monoclonal antibodies. After incubation with a fluorescent probe, the cells were stimulated with various concentrations of PAF, PMA or various cytokines. Both eosinophils and neutrophils generated superoxide dose-dependently in response to PAF and PMA. Eosinophils generated superoxide in response to IL-3 and IL-5 (maximum concentration was 50 ng/ml), but neutrophils did not. IL-5-induced superoxide generation was lower in the presence of IL-5 antibodies. After stimulation with PMA, superoxide generation was significantly higher in neutrophils than in eosinophils (p<0.05). There were no significant differences in PAF-induced or GM-CSF-induced superoxide generation between eosinophils and neutrophils. After stimulation with the optimal concentration of IL-5 or PMA, eosinophils with a lower density generated significantly more superoxide than those with a higher density.

View full abstract

TGF-β1 has many biological activities in various cell types and is regarded as a multifunctioning regulator of cell growth. We studied the effect of TGF-β1 on cytokine-enhanced eosinophil survival in vitro. Eosinophils were purified from patients with mild atopic dermatitis by Percoll density gradient centrifugation and the CD16 negative selection/immunomagnetic beads technique. Eosinophil purity was greater than 95%. The purified eosinophils were incubated in the presence of eosinophil-activating cytokines (IL-5, IL-3, GM-CSF, IFN-γ) with and without TGF-β1 for 3 days. Eosinophl viability was determined by staining the cells with fluorescein diacetate and propidium iodide. Without cytokine, most eosinophils died by day 3 in culture, but human recombinant IL-5, IL-3, GM-CSF, and IFN-y enhanced eosinophil survival in a dose-dependent manner. To test the effect of TGF-β1 on enhanced eosinophil survival, eosinophils were cultured with activating cytokine and TGF-β1. TGF-β1 inhibited eosinophil survival in a dose-dependent manner. The inhibitory effects of TGF-β1 on IL-5 enhanced survival was partially reversed by high concentrations of IL-5 and was completely neutralized with anti-TGF-p antibody. Moreover, the apoptosis of eosinophils induced by TGF-β1 was determined with the assay of DNA fragmentation on agarose gel electrophoresis. It is possible that TGF-β1 activates the pathway of apoptosis. The results suggest that TGF-β1 may play a crucial role in the regulation of allergic inflammation.

View full abstract

IgE is considered tobe involved in immediate hypersensitive reactions (IHR) following egg ingestion. IgE antibody levels to egg-white (EW) antigens in the IHR-positive group (n=19, meanage ± SD=5.2 ± 4.5 yr) were higher than those in the IHR-negative group (n=13, mean of age ± SD = 3.6 ± 2.2 yr). However, even in the IHR-negative group, some patients showed high IgE to EW. RAST inhibition tests with heat-treated (100℃, 5, 10, and 30 min) egg-white antigens were performed on 13 serum samples from subjects with IHR and 9 serum samples from subjects without IHR. Heat treatment decreased the IgE-binding activity of egg white and it was speculated that IgE from IHR-negative subjects bound to relatively heat-unstable sites of egg-white antigens. Furthermore, we selected IHR-negative subjects (n=8, mean of age ± SD = 3.0 ± 1.7 yr) with higher IgE antibody levels than the lowest limit of IgE to EW of the IHR-positive group and compared IgE to ovomucoid (OM), ovalbumin (OA), conalbumin (CA), and lysozyme (Ly) between these IHR-negative and positive groups. IgE-binding activities to egg-white components, including OA, CA, and Ly but not OM, were significantly decreased with heat treatment. The IHR-negative group showed significantly lower IgE to OM (untreated, 5, 10, 30 min treatment) and 5 min treated OA alone than the IHR-positive group, while no difference was found in IgE to other components between the two groups. It was suggested that IgE from IHR-negative subjects with high IgE antibodies to egg-white anitigens might bind to relatively heat-unstable epitopes on egg-white component proteins.

View full abstract

The effect of Sei-hai-to on ion transport in airway epithelial cells and its mechanism of action were investigated. Measurement of short-circuit current (Isc) using cultured epithelial cells from canine tracheal mucosa under short-circuit conditions showed that Isc was dose-dependently increased by the submucosal administration of Sei-hai-to. The response of Isc to this drug was not affected by pretreatment the cells with amiloride, but abolished by furosemide, or Cl-free condition. These results suggest that Sei-hai-to stimulates Cl ion transport selectively and may affect the subsequent movement of water across the airway mucosa to the lumen.

View full abstract

Several kinds of immunological abnormalities have been found more frequently in patients with subacute myelo-optico-neuropathy (SMON). To investigate whether the B-cell immune system is implicated in aging in patients with SMON, we examined serum levels of immunoglobulin includng IgG, IgM, and IgA, and the number of CD20^+ cells (B Iymphocytes) and CD20^+ CD23^+ cells (activated B lymphocytes) using flow cytometry, and compared them with those in age-matched controls. We also investigated whether the number of HLA-DR^+ cells was correlated with those of CD20^+ cells, CD20^+ CD23^+ cells, or HLA-DR^+CD3^+ cells (activated T lymphocytes) in patients with SMON. Serum levels of IgG, IgM and IgA were decreased with aging both in the patients with SMON and in the controls, and no significant difference was found between the two groups. Although the patients with SMON tended to show higher levels of CD20^+ and CD20^+CD23^+ cells than the age-matched controls, there were no significant differences between the two groups. The number of HLA-DR^+ cells was correlated not with that of CD20^+ cells or CD20^+CD23^+ cells, but with that of HLA-DR^+CD3^+ cells. In patients with SMON, it is likely that the B-cell immune system is mainly implicated in the effect of aging, but it is unlikely that other factors than aging are associated with the B-cell immune system. The increase in the number of HLA-DR^+ cells associated with aging in patients with SMON reflects the increase in the number of activated T lymphocytes, and is not correlated with the changes of B lymphocytes.

View full abstract