Role of JAK2 signal transductional pathway in activation and survival of human peripheral eosinophils by interferon-gamma (IFN-gamma).

The purpose of this study was to determine whether the JAK pathway is involved in eosinophil activation and survival through IFN-gamma receptor signalling in human peripheral eosinophils. Eosinophils were purified from the blood of six atopic disease patients by anti-CD16 magnetic bead-negative selection. IFN-gamma significantly up-regulated survival and CD69 expression in 24-48 h cultured eosinophils. Further, IFN-gamma induced tyrosine phosphorylation of JAK2 in eosinophils, as indicated by Western blot analysis. Finally, the specific JAK2 inhibitor AG-490 inhibited the tyrosine phosphorylation of JAK2, IFN-gamma-induced survival and CD69 expression in eosinophils. In conclusion, these results indicate that IFN-gamma induces eosinophil survival and CD69 expression through the activation of JAK2 in peripheral eosinophils, suggesting that JAK2 may play a significant role in eosinophil regulation by IFN-gamma-IFN-gammaR interaction.

Heterogeneous expression of tumor necrosis factor-alpha receptors I and II on human peripheral eosinophils.

BACKGROUND: Expression of the tumor necrosis factor-alpha receptor by human eosinophils has not been determined. We examined the surface expression and mRNA for tumor necrosis factor-alpha receptors I and II (TNF-alphaRI and TNF-alphaRII) on peripheral eosinophils. METHODS: Eosinophils were obtained from 17 asthma patients and 3 healthy volunteers. Flow cytometry was used to examine receptor expression, and the reverse transcriptase-polymerase chain reaction was used to examine receptor mRNA expression. RESULTS: Flow cytometry revealed that 16 out of 20 subjects had eosinophils expressing TNF-alphaRI and 18 subjects had eosinophils expressing TNF-alphaRII. All eosinophils expressed at least one receptor. The mRNA for TNF-alphaRII was detected in all eosinophils examined, but the signals for TNF-alphaRI mRNA were only found in cells expressing this receptor. CONCLUSIONS: Human peripheral eosinophils show heterogeneous expression of TNF-alphaRI and TNF-alphaRII.

Interferon-gamma receptor beta-chain expression and formation of alpha- and beta-chain complexes after receptor conjugation on human peripheral eosinophils.

BACKGROUND: It has been unclear whether or not the interferon-gamma receptor (IFN-gammaR) on human peripheral eosinophils is completely functional. Accordingly, we examined the expression of IFN-gammaR alpha- and beta-chains and the association of the two chains after binding of IFN-gamma to the receptor. METHODS: Peripheral blood eosinophils were obtained from 8 patients (bronchial asthma, n=6, and hypereosinophilic syndrome, n=2), and expression of the alpha- and beta-chain was investigated by flow cytometry with specific antibodies. Expression of mRNA for the alpha- and beta-subunits was also investigated by the reverse transcriptase-polymerase chain reaction. RESULTS: Eosinophils from all the patients were positive for both the alpha- and beta-chains by flow cytometry. In addition, mRNA for both chains was detected by the polymerase chain reaction. Furthermore, coprecipitation of the alpha- and beta-chains was only found after eosinophil stimulation with IFN-gamma. CONCLUSIONS: Human peripheral eosinophils apparently express both the alpha- and beta-chains of the IFN-gammaR, and the two chains may only form the receptor complex after ligand binding.

Human peripheral eosinophils express functional interferon-gamma receptors (IFN-gammaR).

In order to determine whether or not IFN-gammaR is associated with regulatory mechanisms on human eosinophil function, we examined the expression of functional IFN-gammaR on human peripheral eosinophils. In this study, peripheral blood eosinophils were obtained from seven normal controls and 12 patients (bronchial asthma, n = 9, and hypereosinophilic syndrome (HES), n = 3), and the purity of eosinophils was 97.11 +/- 2.31%, n = 19. We first showed that anti-IFN-gammaR alpha-chain MoAb reacted with all tested eosinophils of both normal controls and patients by flow cytometry analysis. We also showed expression of mRNA for the alpha-chain of IFN-gammaR in all purified eosinophils of six individuals. Further, to characterize IFN-gammaR on eosinophils, we did binding experiments with 125I-IFN-gamma on purified peripheral eosinophils. The linear Scatchard plot indicated a single type of high-affinity binding sites (dissociation constant (Kd) = 3.89-4.95 x 10(-10) M, numbers of binding sites = 183-233/cell, n = 3). To determine whether IFN-gammaR on eosinophils is functional, we examined surface eosinophilic cationic protein (ECP) and CD69 induction after IFN-gammaR ligation with recombinant human IFN-gamma (rhIFN-gamma) on eosinophils by flow cytometry. rhIFN-gamma stimulation significantly induced both ECP and CD69 expression on the 2-18 h-cultured eosinophils in a dose-dependent manner. Further, the effects of rhIFN-gamma stimulation were significantly blocked by both a neutralizing anti-IFN-gamma MoAb and a blocking anti-IFN-gammaR MoAb. These results suggest that human peripheral eosinophils express functional IFN-gammaR.

Continuous isoproterenol inhalation therapy in children with severe asthmatic attack.

We studied the 1-type isoproterenol inhalation therapy for patients with severe asthmatic attacks who were admitted at the Department of Allergy of National Children's Hospital from 1981 to 1991. One hour after l-type isoproterenol inhalation therapy, statistically significant effects were noted with regard to the asthmatic status. Moreover, no side effect was found amoung the subjects. From these data, 1-type isoproterenol inhalation therapy is thought to be effective for severe asthmatic attacks.

Study of liver function in infants with atopic dermatitis using the 13C-methacetin breath test.

Serum glutamic-oxaloacetic transaminase (GOT) levels were determined in 214 infants (133 males and 81 females) with atopic dermatitis during their first visit to the Department of Allergy, National Children's Hospital, Tokyo, Japan. Compared with the normal hospital range, their levels were found to be significantly higher, a tendency which was more conspicuous in lower age groups. We carried out a 13C-methacetin breath test (MBT), administering the stable-isotope-labeled compound to 11 children with higher serum GOT values and 5 within the normal range to investigate hepatic metabolism of methacetin in infants with atopic dermatitis. 13C-methacetin was given orally, and the 13CO2 level in the breath was determined immediately before and after administration, by mass spectrometry. Compared to the normal controls, the atopic infants demonstrated significantly lower 13CO2 peak excretion and delayed peak time. The clearance rate of 13CO2 was also decreased. These results suggest some relationship between atopic dermatitis and liver function in infants.

[Does clinical stage have any influence on the results of provocation tests in allergic children? I. The best clinical stage for food provocation tests].

To investigate the best clinical stage for food provocation tests, twenty allergic patients who had early allergic reactions induced by any kind of food were tested. The patients were classified into two groups. Group I; open-food challenge tests were performed on eleven patients within 48 hours of admission. Group II; the tests were performed on nine patients seven or more days after admission. In these food provocation tests, there were nine positive results in Group II (p < 0.05). The percentage of eosinophils in the blood was higher in cases of positive provocation than in cases of negative provocation (p < 0.05). Moreover the percentage of eosinophils decreased after admission day by day in the provocation-negative patients of Group II, but there was no change in the provocation-positive patients. These results suggest that individual sensitivity to food decreased after admission. Therefore, in order to assess the actual sensitivity of allergic children it is necessary that food provocation tests be performed within two days of admission or before admission.

[A case of degenerative type of progressive myoclonus epilepsy].

This report describes a patient with degenerative type of progressive myoclonus epilepsy (PME), who showed slowly progressive deterioration of the central nervous system; intellectual impairment, dysarthria, and involuntary movements, particularly action myoclonus and dystonia. The patient was a 19-year-old woman who had no hereditary factors. At the age of 4, she developed action myoclonus in the upper limbs bilaterally. Her condition became gradually worse, and at the age of 15, she was admitted to our hospital because of involuntary movement in the upper limbs. First physical examination revealed mild mental retardation, action myoclonus, dystonia, and delayed adolescence. As giant SEP characteristic of PME and Ramsay Hunt syndrome was found, she was tentatively diagnosed as having Ramsay Hunt syndrome without epilepsy, and delayed adolescence. Now, she is 19 years old, and unable to walk alone because of involuntary movements and paralysis. But she has not developed epilepsy. As she has not been compatible with progressive myoclonus epilepsy (PME) and progressive myoclonic ataxia (PMA) classified by Marseille Consensus Group, she has been diagnosed as having an atypical PME syndrome.

[Clinical epidemiology of bronchial asthma in children report. No. III. Comparison of prevalence of asthma with 1982 and 1990 and environmental factors].

UNLABELLED: In 1982, our group carried out a survey by the questionnaires and on the site medical check-ups to observe and occurrence of asthma and other allergic symptoms in elementary and junior high school children in Izu Ohshima island. Same procedure was repeated in 1990 for 1145 children of the same age group to see any data fluctuation. The occurrence of allergic symptoms with relation to living conditions was also researched. RESULT: 114 students (11.0%) had either suffered from asthma in the past, or were still currently exhibiting symptoms, this marked a 7.5% rise (108 students) over the 1982 research data. However, patients in need of current treatment numbered 48 (4.5%), which was similar to the 1982 data of 82 (4.3%). For other allergic symptoms, rhinitis showed a marked increase over the 1982 data. The relation between living conditions and asthma was analyzed by Multiple Factor Analysis Quantification Theory. RESULT: 1) Family history of allergy, 2) Selection of food products upon starting solids during infancy, 3) Infection of lower bronchus during infancy all seemed to have had much influence on the occurrence of asthma.

Effects of total asphyxia on the development of synaptic junctions in the brains of mice.

Using ethanolic phosphotungstic acid staining, we studied the development of synaptic junctions in the brains of mice subjected to total asphyxia. One-day-old mice, Std:ddY strain, were put into a chamber continuously flushed with 100% CO2-gas for 35 min. The surviving mice (28.1%) were used as experimental models, while unasphyxiated littermates served as controls. The synaptic junctions in the frontal motor cortex and hippocampal cortex were studied at 20 and 60 days of age using E-PTA staining, and the numbers of synaptic junctions in both areas were counted. In comparison with the control animals, the frontal motor cortex of the subject mice showed a smaller increase in the number of synaptic junctions, both at 20 and at 60 days of age. In the hippocampal cortex, the number of synaptic junctions seen in the experimental mice brains was similar to that in the controls at 20 days of age; however, the number of synaptic junctions in the treated mice brains showed a slower rate of increase than in the controls at 60 days of age.

[Study of IgE.IgG4 antibodies on eczema in infants. II. Clinical characteristics of infants with egg white.RAST 4+].

The author made an assessment in his first report, regarding relationship to the degree of skin involvement. As a result, it was suggested that the extent of eczema is a good index for determining the severity of the allergy and also that the severity of the allergy early in the infancy has a good correlation with egg white.specific IgE antibody. Therefore, an assessment had been made this time regarding the clinical characteristics of 32 infants who have strong reactions (egg white.RAST 4+) to egg white. The results were as follows; 1. All the infants with egg white.RAST 4+ had a systemic eczema, and their IgE antibodies showed high values of 445.6 +/- 626.9 IU/ml. 2. There were many cases with positive multiple allergens. Particularly, all the infants with egg white.RAST 1+ - 2+ were negative to rice and wheat, while 7 cases positive RAST score to rice and 9 cases to wheat were seen out of the 32 who showed the RAST 4+ to egg.white early in the infancy (rice; p less than 0.02 and wheat; p less than 0.01). 3. If the growth curves of the group from which egg and milk were completely eliminated are compared with the curves of the healthy infants, the height and head circumference were entirely in the normal range while the weight and Kaup index had a trend of being a little low, though they were within the normal range. 4. Regarding family history of allergies, past histories of atopic dermatitis on the mother's side were observed at a high rate.

[IgE.IgG4 antibody in eczema in infants between 5-7 months of age. I. Relationship to the degree of eczema involvement].

Assessments have been made of serum IgE and IgA value, specific IgE and IgG4 antibody titers to foods, house dust, mite and percent peripheral++ eosinophil count in 191 infants with and without eczema between 5-7 months of age. Eczema patients were divided into 4 groups according to the degree of eczema involvement and laboratory data were compared between these and not eczematous group. The results were as follows; 1) IgE antibody titers were higher as the degree of eczema involvement increased. 2) RAST positive rates to foods increased with the degree of eczema involvement. Positive rate was highest to egg white followed by milk, soybean, wheat and rice. 3) Only 9 cases were RAST-positive to rice or wheat and all of them had IgE antibodies to other allergens tested. 4) A value of 10 IU/ml, which is the lowest measurable value of IgE in infants was considered to be a little too high to speculate allergy to some foods. 5) Specific IgG4 antibodies were positive only to milk. 6) All the 18 cases with positive milk-specific IgG4 antibody were all negative in milk-specific IgE antibody, and conversely all the 14 cases with positive milk-specific IgE antibody were negative in milk-specific IgG4 antibody. From these results, it was concluded that food allergy is related to the degree of eczema involvement in infants between 5-7 months of age.

[Investigation of the genetics of allergic children, the second report--family study to determine the major gene in allergic disease using affected sib pair method].

Twenty-six families with allergic siblings were investigated to define the mode of inheritance of asthma and DF specific IgE antibody production in children. The affected sib pair method was used to establish the linkage between disease susceptibility genes and HLA antigens. The affected sib pair method revealed that in asthmatic families chi 2 = 4.9 (no significance) and in DF-specific IgE positive families chi 2 = 6.2 (p less than 0.05). This study suggests that the gene of DF-specific IgE antibody production is linked to HLA haplotypes, but the mode of inheritance of DF-specific IgE antibody production couldn't be defined.

[Investigation of the genetics in allergic children: the first report--Twin study on HLA in allergic disease].

Twenty-two twins' families were investigated to determine the influence of a genetic factor in the etiology of allergic diseases in children. Zygosity was confirmed by eight red cell blood types and HLA type. Eight pairs of monozygotic twins were divided by a probability of monozygosity as calculated from Essen-Möller's method. Concordance rate of allergic disease and asthma were 88% in MZ, 86% in DZ, 75%, 73% and that of positive rate of DF-specific IgE antibody were 60% in MZ, 58% in DZ. In DZ, not sharing the same HLA haplotypes, concordance of the positive rate of DF-specific IgE antibody was 0%. Concomitant coefficiency of total serum IgE levels were 0.593 in MZ, 0.063 in DZ. These data suggested that IgE production may be influenced by genetic factors.

[Clinical significance of specific IgG4 antibody in serum].

Specific IgG4 antibodies in sera were measured by ELISA in allergic patients who were diagnosed as susceptible to one or more allergens among mite, milk, soybean or egg white, and also in a non-allergic control group, and their diagnostic significance was investigated. The results obtained were as follows. 1. The level of specific IgG4 antibody was significantly higher in each allergic group than in the non-allergic group. 2. The positive rates in the specific IgG4 antibody determination were higher than those in RAST in the milk-, soybean- and egg white-allergic groups. 3. In each allergic group, the causative allergens were detected more accurately by measuring both specific IgG4 antibody and IgE antibody (RAST) than IgE alone. 4. The positive rates in the specific IgG4 antibody determination were higher than those in the skin test in each allergic group. 5. It was demonstrated that the combination of the skin test with specific IgG4 antibody measurement ensured a more accurate detection of causative allergens than the skin test alone. These results indicated that the measurement of the specific IgG4 antibody is a helpful method to detect the causative allergens in allergic patients.

[A double blind study of the effectiveness of ketotifen in preventing the development of asthma in atopic dermatitis patients].

Many asthmatic children have experienced atopic dermatitis in their younger days. As it is very difficult to cure childhood asthma we attempted to determine the anti-allergic drug effects in preventing the development of asthma by using ketotifen on atopic dermatitis patients. The study was designed as a placebo controlled double blind trial of 128 atopic dermatitis patients aged from 2-34 months. 91 patients were given complete analysis in the study, 33 patients were given only a safety rate and 4 patients were dropped. The 91 patients were followed for 52 weeks. Our primary finding was that the development of bronchial asthma was inhibited in the ketotifen group compared to the placebo controlled group with a statistically significant degree (p less than 0.001). We also found that clinical symptoms of atopic dermatitis were significantly improved in the ketotifen group (p less than 0.001). Only 5 patients complained of mild side effects.

[The study of allergic children under one-year old. The degree of influence on allergic factors to atopic dermatitis by multiple factor analysis].

We investigated the allergic factors of 94 allergic children; 69 atopic dermatitis, 21 wheegy children, 3 urticaria and 1 milk allergy child under one year by multiple factor analysis. The results were as follows; 1) Investigating by relative range ratio the 3 factors, RAST score to Dermatophagoides farinae and egg white, and family history of allergic disease, had strong influence to the existence of atopic dermatitis. 2) By multiple factor analysis type III, the 6 groups had tight relationship to atopic dermatitis group. (1) IgE RIST more than 61 IU/ml group. (2) Eosinophil counts more than 401/mm3 group. (3) RAST score to Dermatophagoides farinae more than 0.70 PRU/ml group. (4) RAST score to egg white more than 0.70 PRU/ml group.

Treatment of status asthmaticus--hormone changes before and after aminophylline i.v. drip therapy.

The treatment of status asthmaticus is one of the most important factors in controlling the patient with asthma attacks. We have studied hormone changes in status asthmaticus and considered what is the best treatment in the asthma attack condition in children. Antidiuretic hormone (ADH), renin activity, and aldosterone activity are elevated in severe asthma attack conditions, and these high levels are correlated with high levels in Wood's clinical score. It is theoretical that patients with dehydration and respiratory failure show such elevation in hormones, and it is well known that under such conditions beta 2-stimulant enhances renin production. From our study, it is concluded that beta 2-stimulant subcutaneous injection must be considered in status asthmaticus. In children, aminophylline i.v. drip therapy may be one of the best treatments in status asthmaticus.