Toshikazu Tsubaki

Gender analysis in acute bronchiolitis due to respiratory syncytial virus

It is reasonable to compare immune reactions between boys and girls because many infections in the early stages are predominant in males. A relationship between immunomodulatory effects of sex hormone surge in boys at early months and infectious diseases is still unclear. We compared clinical features between boys and girls who suffered from wheezing that was initially triggered by acute respiratory syncytial virus (RSV) bronchiolitis. For systemic immune response evaluation, white blood cell (WBC) count, blood eosinophil count, and serum C‐reactive protein (CRP) were measured. For local inflammation evaluation, scores for eosinophils and neutrophils in sputum were evaluated microscopically. Patients consisted of 90 boys and 51 girls. Most children were under 6 months of age. WBC counts and serum CRP levels were significantly increased in girls compared with boys. Blood eosinophilia at the acute stage was rarely observed in children after 6 months of age. For local response evaluation, sputum specimens obtained from 42 boys and 29 girls were microscopically examined. Sputum eosinophil score of 2+ and more was observed in boys (6/42) exclusively. In contrast, sputum neutrophilia was commonly observed in boys and girls. From a follow‐up study, we confirmed that 28 children with RSV bronchiolitis showed wheezing episodes afterwards. However, their blood and sputum eosinophilia during RSV bronchiolitis did not reflect their subsequent wheezing. We speculated that gender‐specific responses to RSV infection might account for male susceptibility. Differences in RSV pathogenecity between boys and girls should be further investigated in terms of asthma progression.

Analysis of sputum taken from wheezy and asthmatic infants and children, with special reference to respiratory infections

Children who are destined to develop asthma are considered to be susceptible to a variety of respiratory pathogens. To elucidate respiratory inflammation among these children, we measured the levels of eosinophil cationic protein (ECP) and tryptase in sputum taken from three different groups of wheezy infants and young children: those with a first wheeze (n = 15); those with recurrent wheeze (n = 27); and those with recurrent wheeze with respiratory distress, namely asthma (n = 56). The numbers of eosinophils or metachromatic cells determined by microscopic analysis of sputum samples were also evaluated in combination with the ECP and tryptase levels. Although neither sputum ECP nor tryptase was a clear discriminative marker that differentiated the three different types of wheezy disease, ECP levels in sputum from the asthma group were significantly higher (2,269.2 ± 6,216.8 ng/g) than those in the recurrent wheezy group (440.3 ± 1,199.8 ng/g) or in the first‐wheeze group (209.0 ± 172.9 ng/g). A similar trend was observed with tryptase levels in sputum, but there were no significant differences among the three groups. Sputum taken from asthmatic children showed a marked accumulation of eosinophils. However, an accumulation of eosinophils in sputum (even in the presence of an elevated level of sputum ECP) was not identified in the asthmatic infants < 1 year of age. An accumlation of eosinophils in sputum was not evident until children became > 1 year old and thereafter the eosinophils rapidly increased in number until the children reached 5 years of age. It was noteworthy that sputa positive for pathogenic bacteria, taken from the 1‐ and 2‐year‐old asthmatic infants, had a tendency to show high levels of ECP but a reduced number of eosinophils. Along with the wheezy episodes induced by viral infection, primarily and occasionally in combination with secondary bacterial infection, eosinophil activation and infiltration may develop. These predestined immune reactions to various pathogens might be associated with triggering the onset of asthma.

Incidence of Latex Allergy in Atopic Children and Hospital Workers in Japan

The incidence of latex-induced allergy has been reported to be increasing in Europe and the US but not in Japan. We thus measured latex-specific IgE antibodies and latex-specific IgG antibodies in sera from 196 atopic children with low serum IgE levels (under 1,000 U/ml; group 1), 108 atopic children with high serum IgE levels (over 1,000 U/ml; group 2) and 601 hospital employees (group 3). Atopic children were diagnosed as having asthma, atopic dermatitis and/or food intolerance. One out of group 1 (0.5%) and 11 out of group 2 (10.2%) were found to have latex-specific IgE by radioallergosorbent assay (RAST), and 7 were further found to be positive for latex antigen by skin test. Fifty-five percent of group 1, 65% of group 2 and 9.7% of group 3 were found to have latex-specific IgG over 100 units/ml by enzyme linked immunosorbent assay (ELISA). Prior to our diagnosis most of the children and employees did not realize they were allergic to latex. These data suggest that caution should be taken regarding latex allergy when atopic children have to be operated upon as atopic children tend to be sensitized to the latex antigen after even minimal contact with latex products.

Bacterial colonization in respiratory secretions from acute and recurrent wheezing infants and children

Lower respiratory tract infection in childhood often results in airway obstruction, characterized by wheezing. However, contribution of bacterial colonization to the wheezy state in children remains unclear. Wheezing and non‐wheezing children requiring hospitalization were classified into three groups: (i) wheezing children having a past history of recurrent wheezing; (ii) wheezing children without such history; and (iii) non‐wheezing children as control subjects. Respiratory secretions as sputum were analyzed microscopically, and cultured. Cultured pathogenic bacterial species in sputum were categorized into two subgroups according to their amounts, i.e., dominant and non‐dominant amounts of colonies. Incidence of bacterial colonization and wheezing were assessed. Hospitalized children were mainly 1‐ to 2‐yr old, and rapidly decreased in number for older ages. Children in the three groups belonged to different clinical entities. Children in the recurrent wheezing group were highly sensitized to mite allergens, and still required hospitalization after 2 yr of age. Incidence of bacterial colonization was similar between the three groups. Dominant and non‐dominant amounts of bacterial colonization were 170/997 (17.1%) and 170/997 (17.1%), respectively, in the recurrent wheezing group; 28/146 (19.2%) and 35/146 (24.0%), respectively, in the acute wheezing group; and 15/56 (26.8%) and 7/56 (12.5%), respectively, in the non‐wheezing group. Regardless of the presence of wheezing, bacterial colonization commonly occurred at a young age in the three groups. In recurrent wheezing children, boys (122/611, 20.0%) carried non‐dominant amounts of bacteria more frequently than girls (48/386, 12.4%) (p < 0.01). Boys showed predominant wheezing and susceptibility to bacterial colonization. Assessment of bacterial colonization allowed us to characterize asthma onset and outgrowth in childhood.

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 Childrens 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.

Growth in Methylcellulose of Human Mast Cells in Hematopoietic Colonies Stimulated by Steel Factor, a c-kit Ligand

We examined the effect of Steel factor (SLF) on the development of human mast cells in hematopoietic colonies from cord blood mononuclear cells in methylcellulose culture. When cord blood cells were cultured for 3 weeks, SLF increased the cellular tryptase levels detected in total cultured cells. It also stimulated the formation of small-cell colonies consisting mainly of polymorphonuclear granulocytes and immature blastoid cells in a concentration-dependent manner but not the formation of colonies consisting of large macrophagic cells. A low percentage of tryptase-positive mast-cell-like cells was found in 39 out of 100 granulocyte/blastoid cell colonies. Four of the 100 colonies contained 10-20% tryptase-positive cells, but we failed to observe colonies consisting of > 20% of tryptase-positive cells. These results suggest that the effect of SLF on mast cell growth is brought on by stimulating the growth of primitive hematopoietic progenitors.

Age and sex as factors of response to RSV infections among those with previous history of wheezing

Although enhanced immune reaction caused by the respiratory syncytial virus (RSV) in allergen‐sensitized animal model has been reported, RSV illnesses in children already sensitized or having recurrent wheezing episodes have not been completely studied. In addition, the reason for male dominances in RSV infection at young ages was also inconclusive. Therefore, gender analysis in recurrent wheezing children with RSV infection can shed light on asthma pathogenesis. We studied the clinical features and the laboratory data of RSV infections in children who had recurrent wheezing histories. The subjects with RSV infection consisted of 98 boys and 58 girls. The children under 4 yr of age were 123 (78.8%) in number. Children with pneumonia were 78 and those with febrile episode were 119. Children above 1 yr of age were highly sensitized with mite antigen (75/96, 78.1%). The clinical symptoms and signs differed according to their ages. Children in each age group behaved differently in their immune reaction to RSV. Above all, 3‐yr‐old children deteriorated clinically during acute RSV infection, accompanied by transient elevated C‐reactive protein (CRP) and suppressed blood eosinophil counts. Clinical features differed in several points between boys and girls. In general, the white blood cell count and the CRP levels were higher in girls in every age group. Blood eosinophil counts at the acute illness were significantly higher in boys than girls aged 2 and 3< yr. Age and gender comparison in already sensitized children might suggest a clue to asthma pathogenesis.

Changes in Histamine/Tryptase Levels in Skin Chambers: Application for Clinical Evaluation of Atopic Dermatitis

We measured changes in histamine and tryptase levels in fluid of children with atopic dermatitis using skin chamber methods and evaluated the correlation with clinical symptoms. Skin chambers were app

Pranlukast reduces asthma exacerbations during autumn especially in 1- to 5-year-old boys

Background Leukotriene receptor antagonists have been used to prevent virus-induced asthma exacerbations in autumn. Its efficacy, however, might differ with age and sex. Objective This study aimed to investigate whether pranlukast added to usual asthma therapy in Japanese children during autumn, season associated with the peak of asthma, reduces asthma exacerbations. It was also evaluated the effect of age and sex on pranlukasts efficacy. Methods A total of 121 asthmatic children aged 1 to 14 years were randomly assigned to receive regular pranlukast or not according to sex, and were divided in 2 age groups, 1–5 years and 6–14 years. The primary outcome was total asthma score calculated during 8 weeks by using a sticker calendar related to the days in which a child experienced a worsening of asthma symptoms. This open study lasted 60 days from September 15 to November 14, 2007. Results Significant differences in pranlukast efficacy were observed between sex and age groups. Boys aged 1 to 5 years had the lower total asthma score at 8 weeks (p = 0.002), and experienced fewer cold episodes (p = 0.007). There were no significant differences between pranlukast and control group in total asthma score at 8 weeks (p = 0.35), and in the days in which a child experienced a worsening of asthma symptoms (p = 0.67). Conclusion There was a substantial benefit of adding pranlukast to usual therapy in asthmatic children, especially in boys aged 1 to 5 years, during autumn season.

Airway responses to repeated exercises detected by krypton-81m in asthmatic children.

A repeated exercise program was used to test 7 asthmatic children for changes in ventilation. These changes were examined by continuously inhaled Krypton-81m and compared in subjects with positive and negative refractoriness, as defined by forced expiratory volume in 1 second (FEV1). Three of the seven patients showed significant refractoriness (% reduction in FEV1 > 50%). After the first exercise, they showed one or two ventilation defects which improved after the second exercise. The patient with incomplete refractoriness showed similar results. On the contrary, subjects without refractoriness showed several ventilation defects which fluctuated after the second or third exercise program. One defected area improved after the second exercise session, but deteriorated after the third; and another area deteriorated after the second exercise and improved after the third. It was concluded that approximately half of the patients were refractory in view of FEV1, but that they were all refractory in view of regional ventilation 81mKr images.