Non-atopic Group Research Articles

STAT1 phosphorylation and cleavage is regulated by the histamine (H4) receptor in human atopic and non-atopic lymphocytes

Histamine can modulate the balance between T helper lymphocytes 1 and 2 (Th1 and Th2), and there is evidence that allergic reactions can be associated with excessive histamine production causing shifts toward Th2 responses. As signal transduction in Th-cells is specifically correlated to signal transducer and activator of transcription (STAT) activation and expression, the question arises whether histamine acting through histamine receptors (HR) induces modulation of the Janus kinase (JAK)-STAT pathway. Peripheral blood mononuclear cells (PBMC) from atopic and non-atopic donors were stimulated with phytohemagglutinin (PHA). Initial interleukin-4 (IL-4) levels were higher in the atopic group compared to the non-atopics and interferon-γ (IFN-γ) levels were lower. This was correlated to lower levels of STAT1 expression and phosphorylation. Furthermore, Western blots showed a 118-kDa STAT1 band at the start of the PHA stimulation that was apparently cleaved to STAT1α (91 kDa) and a 28 kDa-fragment with further stimulation. Histamine or the H4R agonist, clobenpropit, led to a significant suppression of the formation and phosphorylation of STAT1α in the non-atopic group after 48 h of PHA stimulation, but had no effect in the atopic group where STAT1α levels were already reduced. The STAT1α levels in the non-atopic group were enhanced by the H4R antagonist JNJ7777120. The phosphorylation of STAT1 could also be potentiated by the H4R antagonist, mimicking the precursor responses of STAT1α. JNJ7777120 also increased the binding of STAT1 to DNA and this effect could be reversed by histamine. As for histamine, the effects of the H4R antagonist were only seen in the non-atopic group. These results suggest that, in non-atopic individuals, histamine acting via the H4R can influence STAT1 regulation, but that this pathway is not present in atopics perhaps due to constitutive suppression of STAT1 activity.

Suppression of IFN‐gamma production in atopic group at the acute phase of RSV infection

Several studies have suggested that respiratory syncytial virus (RSV) bronchiolitis induced the change of cytokine production profile in childhood. We sought to determine whether the RSV-induced cytokine production was affected by the patient's atopic background. We quantified interferon-gamma (IFN-gamma) and interleukin (IL)-4 in the supernatant of peripheral blood mononuclear cells (PBMCs) cultured for 24 h and in the presence of phytohemaglutinin (PHA), IL-12, or IL-18, from 14 infants who were divided into two groups, those who are non-atopic and an atopic group. In RSV-infected infants with atopic diseases, IFN-gamma production from IL-12- or especially IL-18-stimulated PBMCs was subtotally suppressed in the acute phase, whereas in RSV-infected infants without atopic diseases IFN-gamma production was not suppressed on acute phase. The IFN-gamma suppression observed in the atopic group is not caused by the immaturity of an infant's immune system since reduced IFN-gamma production to RSV is not observed in the infants of non-atopic group. IFN-gamma suppression in regard to RSV infection might be caused by some genetic factor involved in the development of atopic disease such as IL-18 signal cascade.

Repercussão da imunoterapia específica na população T1 e T2 de linfócitos periféricos em doentes atópicos

Allergen-specific immunotherapy has been used for successful treatment of atopic diseases. They may act by modifying the patterns of cytokines produced by T cells. However, the precise mechanism by which it accomplishes these effects is still incompletely understood.Objective: To evaluate the effect of one year immunotherapy on cytokines profiles T1 and T2 of peripheral blood lymphocytes in atopic patients.Methods: We studied 10 atopic patients sensitised to common environmental allergens receiving immunotherapy over one year mean period. Six of these patients were studied before and after immunotherapy. Fourteen atopic patients untreated and 7 non-atopic subjects were used as control groups. Intracellular cytokine production (IFN-γ; IL-4; IL-5; IL-10) was determined by flow cytometry following stimulation with phorbol myristate acetate (PMA), ionomycin and brefeldin. Mann-Whitney U and Wilcoxon non-parametric tests were utilized for the statistical analysis.Results: The expression of IL-4 and IL-5 in T cells, characteristically increased in atopic patients, respectively 13.8 (3.1 – 31.8) and 6.7% (1.0 -20.4), was significantly lower in the immunotherapy group [5.4 (2.9 -15.6) p=0.007 and 2.1% (0,6 – 4.8) p=0.035] and similar in the non-atopic control group. The levels of IFN-γ did not differ between the studied groups but the ratio IFN-γ / IL-4 produced by CD4+ T lymphocytes increased significantly in the patients receiving immunotherapy. In addition, there was an increase in the expression of IL-10 by T cells of the immunotherapy group compared to the non-atopic controls [1.9 (1.0 – 4.9) versus 1.4% (0.9 – 1.4) p=0.02], being more evident in CD8+ T lymphocytes. IL-10 correlated significantly with all the profile T2 cytokines (IL-4 and IL-5) and with the phenotype Tc2.Conclusion: After one year of immunotherapy the peripheral T cells response to a polyclonal stimulation revealed a reduction in IL-4 and IL-5 production, characteristically increased in atopic disease. The increase of IL-10 that we found in our study suggested the existence of a profile T2 regulatory population, more evident in CD8+T lymphocytes.Rev Port Pneumol 2006; XII (2): 107-130

Specific immunotherapy effect on peripheral blood T1/T2 lymphocytes in atopic patients

Allergen-specific immunotherapy has been used for successful treatment of atopic diseases. They may act by modifying the patterns of cytokines produced by T cells. However, the precise mechanism by which it accomplishes these effects is still incompletely understood. ObjectiveTo evaluate the effect of one year immunotherapy on cytokines profiles T1 and T2 of peripheral blood lymphocytes in atopic patients. MethodsWe studied 10 atopic patients sensitised to common environmental allergens receiving immunotherapy over one year mean period. Six of these patients were studied before and after immunotherapy. Fourteen atopic patients untreated and 7 non-atopic subjects were used as control groups. Intracellular cytokine production (IFN-γ; IL-4; IL- 5; IL-10) was determined by flow cytometry following stimulation with phorbol myristate acetate (PMA), ionomycin and brefeldin. Mann-Whitney U and Wilcoxon non-parametric tests were utilized for the statistical analysis. ResultsThe expression of IL-4 and IL-5 in T cells, characteristically increased in atopic patients, respectively 13.8 (3.1 – 31.8) and 6.7% (1.0 -20.4), was significantly lower in the immunotherapy group 5.4 (2.9 -15.6) p=0.007 and 2.1% (0,6 – 4.8) p=0.035 and similar in the non-atopic control group. The levels of IFN-γ did not differ between the studied groups but the ratio IFN-γ / IL-4 produced by CD4+ T lymphocytes increased significantly in the patients receiving immunotherapy. In addition, there was an increase in the expression of IL-10 by T cells of the immunotherapy group compared to the non-atopic controls 1.9 (1.0 – 4.9) versus 1.4% (0.9 – 1.4) p=0.02, being more evident in CD8+ T lymphocytes. IL-10 correlated significantly with all the profile T2 cytokines (IL-4 and IL-5) and with the phenotype Tc2. ConclusionAfter one year of immunotherapy the peripheral T cells response to a polyclonal stimulation revealed a reduction in IL-4 and IL-5 production, characteristically increased in atopic disease. The increase of IL-10 that we found in our study suggested the existence of a profile T2 regulatory population, more evident in CD8+T lymphocytes.

Eosinophil infiltration of the oesophageal mucosa in patients with pollen allergy during the season

The oesophagus is normally devoid of eosinophils. There are some disorders accompanying with eosinophil infiltration. Food allergy has been reported as a common reason, especially in children but some other studies have also indicated that aeroallergens might have a role in oesophageal eosinophil accumulation. In this study we investigated whether there is any eosinophil recruitment in the oesophagus of pollen-allergic patients who had respiratory symptoms during the season. Thirty-eight symptomatic patients (allergic rhinitis (AR) with or without asthma) who had sensitization to grass pollen were included in the study during the pollen season. Controls were composed of 25 healthy non-atopics and 24 patients diagnosed as having gastro-oesophageal reflux disease. Reflux was excluded in allergic and non-atopic groups, whereas the presence of allergy was eliminated in control groups. Gastrointestinal endoscopy was performed in all participants, and biopsy specimens were taken from both the proximal and the distal oesophagus to evaluate eosinophil accumulation. At the same time, blood eosinophil numbers were counted. Results Oesophageal eosinophil accumulation was found in 10 allergic patients (26%) and in five patients (21%) with gastro-oesophageal reflux disease but none of the healthy controls had eosinophils (0%) (P<0.05). Blood eosinophils were higher in these 10 patients than the rest of the 28 patients without infiltration. In this group, blood eosinophils were also correlated with the number of accumulated eosinophils in the oesophagus (P<0.001). There was more intense eosinophil infiltration at the distal part of the oesophagus in the reflux group when compared with the allergic group (mean 7.6+/-5.6 vs. 3.2+/-3.7). Nevertheless, eosinophils were found to be concentrated (mean 5.5+/-7.3) in the proximal oesophagus of allergic patients, although it was 1.7+/-1.5 in reflux patients (P>0.05). Conclusion Our results showed that eosinophil infiltration might be observed in oesophageal tissue of patients with respiratory tract allergy during the symptomatic period. This finding may possibly reflect the systemic and common mucosal aspects of allergic inflammation.

Exhaled nitric oxide and exercise‐induced bronchoconstriction in young male conscripts: association only in atopics

This study was conducted to evaluate how bronchial responsiveness to direct and indirect stimuli relate to nitric oxide producing airway inflammation, and whether the relationship differs between atopic and nonatopic patients with various degrees of bronchial hyperresponsiveness and airway inflammation in a group of otherwise homogenous young men. We studied 181 consecutive non-smoking steroid-naive young male conscripts referred to military hospital because of respiratory symptoms suggesting asthma. Skin prick tests, spirometry, measurement of exhaled nitric oxide (FENO), and standardized airway challenges with histamine and exercise were performed. 128 patients were atopic. FENO was significantly higher in the atopic group, median 21.2 ppb, compared to 10.2 ppb in the nonatopic group. Still, 36% of all nonatopic patients had elevated FENO. Bronchial responsiveness to histamine (HIB) was similar in the two groups, but exercise-induced bronchoconstriction (EIB) was stronger in atopics (P < 0.01). FENO associated significantly with atopy (P < 0.001), severity of EIB (P < 0.001) and HIB (P = 0.006) in multiple linear regression model. In separate regression models for atopic and nonatopic patients FENO associated with severity of EIB and HIB in atopic patients only. The results were similar when patients with confirmed diagnosis of asthma were analyzed separately. Our results indicate that FENO significantly associates with EIB and HIB in atopic, but not in nonatopic steroid-naïve patients with asthmatic symptoms. The finding suggests that in such atopic patients degree of airway hyperresponsiveness may reflect severity of airway inflammation. However, in nonatopic patients with similar symptoms other mechanisms of airway hyperresponsiveness may be more important.

A different pattern of risk factors for atopic and non‐atopic wheezing in 9–12‐year‐old children

Few epidemiological studies have compared the risk factors of asthma or wheezing between atopic and non-atopic children. The objective of this study was to determine if there are specific risk factors for current wheezing related to atopic status in schoolchildren. Schoolchildren 9-12 yr of age from three Spanish cities (n = 2720) were subject to a cross-sectional study of asthma risk factors (by questionnaire) and atopy (by skin prick test) according to the ISAAC phase-II protocol. Risk factors for current wheezing (in the last 12 months) as reported by parents were investigated among the atopic (positive prick test to at least one allergen) and the non-atopic (negative prick test) children. The prevalence of current wheezing was 13.1% in the whole group, 22.1% in the atopic group and 7.8% in the non-atopic group. However, only 62.4% of children with current wheezing were atopic. Male gender and asthma in the mother and/or the father were both significant and independent risk factors for current atopic wheezing, whereas maternal smoking in the first year of the child's life and mould stains on the household walls were for current non-atopic wheezing. In summary, this study shows that atopic and current non-atopic wheezing children in Spain do not share identical environmental and family risk factors.

Effects of nebulized sodium cromoglycate on adult patients with severe refractory asthma

Many patients with severe refractory asthma, which is insufficiently controlled by additional high-dose of inhaled corticosteroids, require oral corticosteroids and/or immunosuppressant. Clinicians should seek for suitable medications, for its' chronic use may induce high risk of side effects. The purpose of this study was to evaluate the efficacy and safety of nebulized sodium cromoglycate (3-4 times/day) in adult severe asthmatic patients with poorly controlled asthmatic symptoms, despite treatments with high-dose inhaled corticosteroids. Adult patients with severe asthma (n=251) were enrolled in a randomized clinical trial at 30 medical centers in Japan. Isotonic saline was used as placebo. The study was conducted for 10 weeks; with initial 2 weeks of observation followed by 8 weeks of treatments. Efficacy was primarily evaluated based on improvements in morning peak expiratory flow after treatment. All patients who applied inhalation of nebulized sodium cromoglycate (SCG group) or saline (Controls) were treated with high-dose of inhaled corticosteroids (median of beclomethasone dipropionate equivalent dose: 1600 microg/days) and second-line control therapy including oral corticosteroids. There was no significant difference in morning peak expiratory flow between SCG group and controls. However, when patients were stratified into atopic and non-atopic groups, morning peak expiratory flow had significantly improved in the atopic SCG group compared to atopic Controls. Additional inhalation of nebulized sodium cromoglycate with inhaled corticosteroids is effective even in patients with severe atopic asthma. This finding shows that nebulized sodium cromoglycate is expected to be new second-line therapeutic option in severe asthma.

Th1 and Th2 Cell Population in Chronic Ethmoidal Rhinosinusitis: A Chemokine Receptor Assay

It is necessary for the immune system in the nasal and paranasal sinus mucosa to possess appropriate responses to infective agents as well as allergic responses. Recently, the T helper (Th)1/Th2 paradigm has been proposed as a new concept to explain various immunologic phenomena. Toward the goal of better understanding chronic rhinosinusitis, we have tried to define the predominating helper T-cell subsets among patients with chronic rhinosinusitis through characterizing the expressed chemokine receptors by these cells to find out the relation between these chemokine receptors' expression and the underlying pathogenesis of chronic rhinosinusitis. Thirty patients with ethmoidal chronic rhinosinusitis were used in our study. Patients were divided into atopic and nonatopic groups according to their serum immunoglobulin (Ig)E levels. Samples from ethmoidal sinus mucosa were processed as follows: frozen sections were examined immunohistochemically for detection of CCR4, CCR5, and EG2 positive cells and the mRNA of CCR4, and CCR5 transcripts were then examined by real-time quantitative polymerase chain reaction (PCR). By immunohistochemistry, atopic patients showed high expression of CCR4 and EG2 positive cells, whereas nonatopic patients showed high expression of CCR5 positive cells. The expression of CCR4 and CCR5 mRNA, detected by real-time quantitative PCR, supported the data obtained by immunohistochemistry. The present study suggests that eosinophil recruitment associated with Th2 cell infiltration is the main factor responsible for the pathology of atopic rhinosinusitis.

Evidence for two independent distributions of serum immunoglobulin E in atopic families: cognate and non-cognate IgE

Genetic studies of IgE-mediated atopic disease have produced conflicting results, due largely to variable phenotype definitions. Total IgE concentrations and 14 allergen skin prick tests (SPT) were determined in 1099 members of families with history of atopy. Log 10 [Total IgE] values were normally distributed in both atopic (SPT [+]) and non-atopic (SPT [−]) groups. The mean Log 10 [Total IgE] value was higher in the atopic group, although the standard deviations of the distributions were the same. The mean Log 10 [Total IgE] value of the non-atopic distribution was subtracted from the individual Log 10 [Total IgE] values of the atopic group giving an allergen-specific fraction. There was a strong positive correlation between the specific IgE fraction and the number (#) SPT [+] results, defined as Cognate IgE. Among the atopics, subtracting the Cognate IgE value from total IgE yielded Non-Cognate IgE. The Cognate and Non-Cognate IgE distributions were statistically uncorrelated. Evidence is presented for two serum IgE fractions that are statistically and physiologically independent of one another in atopic families; a Cognate IgE fraction associated with atopic sensitization and a Non-Cognate IgE fraction unrelated to atopic disease. Elevated serum IgE is a consequence, not a predisposing cause, of allergen sensitization.

Ex vivo induction of cytokines by mould components in whole blood of atopic and non-atopic volunteers.

We investigated the time-course release of IL-1beta and IL-8 protein as well as the steady state mRNA level of their genes in human whole blood after stimulation with LPS, beta-1,3-D-glucan and mould extracts. We compared the response of 10 non-atopic and 10 atopic individuals. In parallel, cytokine protein release and the corresponding steady state mRNA level was determined by the standard ELISA and real-time on-line RT-PCR methods, respectively. Glucan induced the highest level of IL-1beta mRNA and protein release after 3 h. IL-8 was induced at 3 h after glucan, but not after LPS, induction, which indicates different induction pathways. At the time-points found to elicit the optimal cytokine response significantly higher basal cytokine mRNA levels and significantly lower LPS induced cytokine mRNA levels were observed in the non-atopic group. Generally, mould components induced cytokine mRNA steady state to lower levels in the atopics compared to the non-atopics. In contrast, no differences were found between the two groups in their capacity to induce cytokine protein release. These findings persisted after correction for the percentage of mononuclear cells. The data supported our hypothesis that "exhaustion" of the atopic immune system elicits lower basal cytokine mRNA levels and that analysis of cytokine gene expression has the potential to differentiate between non-atopic and atopic individuals.

Elevation of plasma transforming growth factor β1 levels in stable nonatopic asthma

Increased transforming growth factor beta1 (TGF-beta1) levels have been reported in bronchoalveolar lavage fluid and bronchial biopsy specimens from asthmatic patients. However, systemic TGF-beta1 levels have not been reported in asthma. To evaluate the levels of plasma TGF-beta1 in asthmatic patients and matched, healthy controls to determine the associations with atopic status, disease severity, and duration. Asthmatic patients and healthy controls were recruited prospectively from a university hospital outpatient department between January 2001 and May 2002. Plasma TGF-beta1 and serum IgE levels were estimated using established methods. Patients were classified as atopic or nonatopic based on the presence or absence of serum specific IgE directed to common allergens. Of the 56 patients recruited for the study, 32 were atopic and 24 nonatopic. The median value of plasma TGF-beta1 was significantly higher in nonatopic asthmatic patients (2.5 ng/mL) compared with controls (1.5 ng/mL, P = .002) and atopic asthmatic patients (1.4 ng/mL, P = .008). The median absolute neutrophil count in the nonatopic asthmatic patients (4.0 x 10(9)/L) was significantly higher compared with atopic asthmatic patients (3.0 x 10(9)/L) and healthy controls (3.5 x 10(9)/L) (P = .01 and P = .04). There was no significant correlation between the duration or severity of asthma and plasma TGF-beta1 levels. The distribution of moderate-persistent asthma cases was similar in atopic and nonatopic groups. Compared with atopic asthmatic patients and healthy controls, patients with nonatopic asthma have elevated plasma TGF-beta1 levels and leukocytosis. These data suggest that nonatopic asthmatic patients exhibit an altered inflammatory response, perhaps to a respiratory infection.

INCREASED LEUKOTRIENES IN EXHALED BREATH CONDENSATE IN CHILDHOOD ASTHMA

Csoma Z, Kharitonov SA, Balint B, Bush A, Wilson NM, Barnes PJ. Am J Respir Crit Care Med. 2002;166:1345–1349To determine if 2 markers of airway inflammation, leukotrieneB4 (LTB4) and cysteinyl leukotrienes (cys-LTs) can be detected in the breath of children with asthma and to look at the effect of steroid treatment on these mediators.There were 4 groups of children all between the ages of 7 and 14 years. In addition to the healthy nonatopic control group (n = 11), there were children with mild intermittent asthma (n = 11), mild persistent asthma (n = 13), and a group that contained both moderate and severe persistent asthma (n = 13). The diagnosis of asthma was based on American Thoracic Society criteria. Severity classifications followed the National Heart Lung and Blood Institute/World Health Organization guidelines and the diagnosis of allergy was based on skin test responses to common allergens. Each study group was similar in age and ratio of male/female except there were considerably more boys in the moderate-severe group. The mild persistent group was on <400 μg/day of inhaled steroids whereas the more severe group was divided between those on >400 and those on ≥1000 μg/day. No one was on a leukotriene modifier. The study design involved clinical history, spirometry, and measurements of nitric oxide (NO) and leukotrienes in the breath condensate.Cys-LT was detectable in the exhaled breath of normal, nonasthmatic, nonatopic children (18.5 ± 0.5 pg/mL). Levels were significantly increased in children with mild persistent asthma (27.9 ± 2.8 pg/mL) and moderate/severe persistent asthma (31.5 ± 4.5 pg/mL). Cys-LT levels in exhaled breath of children with mild intermittent asthma were similar to the control group (19.9 ± 1.1pg/mL). LTB4 levels were significantly increased in the breath condensates of children with mild persistent asthma (126 ± 8.8 pg/mL) and moderate/severe persistent asthma (131.9 ± 7.1 pg/mL) as compared with mild intermittent asthma (52.7 ± 3.8 pg/mL) and the control subjects (47.9 ± 4.1 pg/mL). In patients with mild persistent asthma, there was an inverse correlation between levels of cys-LT and LTB4. With increasing amounts of cys-LT there was a decrease in the amount of LT B4. The amount of exhaled NO was increased significantly in all asthmatic children compared with controls.Two markers of inflammation, cys-LT and LTB4, were found to be elevated in the exhaled breath of children with mild to moderate/severe persistent asthma. These mediators were found in those already on inhaled corticosteroids. The findings support the involvement of these 2 leukotriene mediators in chronic airway inflammation. This study also shows that exhaled breath condensate may be useful in assessing inflammation in the airways of children 6 to 7 years old.For many years now we have been enthusiastic supporters of the concept that asthma is a chronic inflammatory disease of the airway. Yet, as these authors point out, we have not been able to measure what we think is important in children. Previous work on airway inflammation has involved adults undergoing rather invasive diagnostic techniques. This study introduces a noninvasive measurement of inflammation and although in small numbers clearly shows differences in inflammatory mediators in the various classifications of disease. The cys-LT are derived from mast cells and eosinophils and include mediators that are blocked by available leukotriene modifiers. All persistent forms of asthma that were treated with inhaled corticosteroids showed elevations of cys-LTs. LTB4 is derived from neutrophils. This neutrophil product is also elevated in the persistent forms of asthma. One of the take-home messages from this work is the fact that despite the use of the inhaled steroids, by-products of inflammation were detected in these children who were asymptomatic. There is also the concern or argument that deals with the presence of inflammation in all forms of asthma—the intermittent and persistent. The measures taken here did not show any of the inflammatory leukotriene mediators and only a slight increase in NO. We may have observed the introduction of a noninvasive tool to help understand the process of airway inflammation and perhaps guide therapy.

Influence of sensitization to inhalative allergens on adenotonsillar disease.

We sought to determine the influence of IgE-mediated sensitization on adenotonsillar disease in children. We compared follow-up after tonsillectomy/adenoidectomy of atopic and nonatopic children.Study design and setting A prospective study of 293 children consecutively undergoing tonsillectomy/adenoidectomy was conducted at a university hospital center. Preoperative and postoperative (1-year follow-up) allergy-related symptom scores were obtained by parents. Intraoperative total serum IgE, the screening test sx1 (Pharmacia), and, if positive, serum specific IgE to inhalative allergens were carried out. Sixty-seven children (22.9%) showed positive RAST results to inhalative allergens (class 1 to 6). In both sensitized and nonsensitized groups, the general health was improved in more than 89% at 1 year postoperatively. No significant difference of postoperative nasal symptoms between the atopic and nonatopic groups was found. Both atopic and nonatopic children with adenotonsillar disease improve health after adenoidectomy or adenotonsillectomy. In our opinion, routine allergy tests before adenotonsillectomy or adenoidectomy are not justified. Only if allergy is suspected due to clinical findings or family history should an allergy test be carried out.

Influence of sensitization to inhalative allergens on adenotonsillar disease

We sought to determine the influence of IgE-mediated sensitization on adenotonsillar disease in children. We compared follow-up after tonsillectomy/adenoidectomy of atopic and nonatopic children.Study design and setting A prospective study of 293 children consecutively undergoing tonsillectomy/adenoidectomy was conducted at a university hospital center. Preoperative and postoperative (1-year follow-up) allergy-related symptom scores were obtained by parents. Intraoperative total serum IgE, the screening test sx1 (Pharmacia), and, if positive, serum specific IgE to inhalative allergens were carried out. Sixty-seven children (22.9%) showed positive RAST results to inhalative allergens (class 1 to 6). In both sensitized and nonsensitized groups, the general health was improved in more than 89% at 1 year postoperatively. No significant difference of postoperative nasal symptoms between the atopic and nonatopic groups was found. Both atopic and nonatopic children with adenotonsillar disease improve health after adenoidectomy or adenotonsillectomy. In our opinion, routine allergy tests before adenotonsillectomy or adenoidectomy are not justified. Only if allergy is suspected due to clinical findings or family history should an allergy test be carried out.

Dysregulation of IL-13 production by cord blood CD4+ T cells is associated with the subsequent development of atopic disease in infants.

Early intervention strategies in allergic diseases will be dependent on identification of newborns at high risk for later development of atopic disease. In this cohort study of 106 neonates, we investigated whether cytokine production property and responsiveness to IL-12 of neonatal CD4(+) T cells were associated with the subsequent development of atopic disease and whether a skewed cytokine production property was intrinsic to helper T cells. To exclude the effects of contaminating cells, highly purified cord blood CD4(+) T cells were stimulated with anti-CD3 MAb and recombinant B7-2 molecule in the presence or absence of IL-12. Production of IL-13 and interferon-gamma was determined by ELISA. The infants were assessed at 12 mo for the development of atopic diseases. CD4(+) T cells of neonates who manifested allergic symptoms (atopic group) produced higher levels of IL-13 compared with those of the nonatopic group in both the presence and absence of IL-12. No significant difference was noted between the two groups with respect to interferon-gamma production. Moreover, higher IL-13 production was also observed in neonates with chronic eczema than those with short-term eczema. Our data suggest that increased production of IL-13 by neonatal CD4(+) T cells is a useful marker of newborns at high risk for subsequent development of atopic diseases and that an intrinsic abnormality of CD4(+) T cell is associated with the pathogeneses of atopic disease, especially atopic dermatitis in infants.

The Impact of in Vivo Calmette-Guérin Bacillus Administration on in Vitro IgE Secretion in Atopic Children

To investigate whether a preexisting T helper (TH) 2 type immune response could be suppressed by Calmette-Guérin Bacillus (BCG) immunization in atopic children with asthma, we determined interferon (IFN)-γ, interleukin (IL)-2, IL-4, and IL-5 and total IgE level in the supernatant of peripheral blood mononuclear cells (PBMC) of six atopic and five nonatopic children in response to phytohemagglutinin A (PHA), purified protein derivate (PPD), and Dermatophagoides pteronyssinus II allergen (Der p II) both before and after BCG vaccination. IL-5 level in response to Der p II was significantly higher in the atopic group than in the nonatopic group both before and after BCG vaccination (p = 0.004, p = 0.009, respectively). In the atopic group, IgE levels determined in PPD and Der p II stimulated and unstimulated culture supernatants decreased significantly after BCG vaccination (p = 0.028, p = 0.026, p = 0.046, respectively), whereas in the nonatopic group (p = 0.041) BCG vaccination resulted in a significant decrease in IgE level only in response to Der p II stimulation. We concluded that in vivo BCG administration can downregulate both spontaneous and stimulated in vitro IgE secretion from PBMC of atopic children.

IL-10 inhibits allergen-specific TH2 cytokine production by human peripheral blood mononuclear cells from patients with ragweed allergies

~ A IL-10 Inhibits Allergen-Specific TH2 Cytokine Production by 1 ~jl-lr Human Peripheral Blood Mononuclear Cells From Patients With Ragweed Allergies J Sun, Susanna Goncharova, M Conway, A Dalrymple, Elizabeth C Cates, Martin R Stampfli, B Wong, Susan Waserman, Manel Jordana McMaster University, Hamilton, ON, Canada Type 2 polarized T cells and their effector cytokines play a central role in allergic diseases. The objective of this study was to investigate whether ILl0, a cytokine with powerful immuno-suppressive activities, inhibits Th2 cytokine production by peripheral blood mononuclear cells (PBMC) from patients with ragweed allergies. The patients were assessed by skin prick test and clinical history, and PBMC were isolated from these ragweed allergic individuals and non-atopic controls prior to pollen season. PBMC were stimulated with a ragweed pollen extract and cytokine synthesis was measured in culture supematants. Spontaneous production of IL-5 and IL-13 varied with each individual and was not significantly different between the atopic and non-atopic group. In ragweed allergic patients, addition of ragweed extract to the cell culture markedly enhanced levels of IL-5 and IL-13. In contrast, PBMC from non-atopic controls did not respond to ragweed stimulation. Both spontaneous and allergen-induced IL-5 and IL13 production were significantly inhibited by recombinant human IL10. IL10 also suppressed Th2 cytokine production induced by PMA, a T cell polyclonal stimulator. Interestingly, IL-4 synthesis could not be detected in any of the samples tested. We are currently assessing the impact of IL-10 on chemokine production. We also extracted RNA from cultured cells at 3h, 6h, 12h, 24 h, 72h, and 5 days and are assessing the expression of chemokine receptors and co-stimulatory molecules using real time quantitative PCR. Our findings in samples collected prior to pollen season will be compared to samples being generated during pollen season. In summary, PBMC from ragweed allergic individuals produce significant levels of IL-5 and IL-13 in response to ragweed stimulation. This response is inhibited by exogenous IL-10. Thus, our results further the notion that IL10 has therapeutic potential in allergic diseases.

Risk Factors Determining Allergic Airway Diseases in Turkish Subjects

With regard to changes in life-style after the 1980s, the reflection of environmental factors on the evolution of allergic airway disease (AAD) has not yet been analyzed in Turkey. In this trial, we aimed to determine the factors related to AAD in Turkish subjects. Patients diagnosed as having asthma and/or rhinitis with positive skin prick tests (SPTs) were accepted as members of the “atopic group” (n = 235). Subjects demonstrated to have negative SPTs and no clinical evidence of any atopic disorder were accepted as members of the “nonatopic control group” (n = 84). Data obtained from a questionnaire applied cross-sectionally to the study groups were compared for risk factor analysis. According to the results of univariate analysis, nonatopic controls were found to have been born in villages more frequently than those in the atopic group (14.3% vs. 4.7%, p= 0.02). Atopic group members had lived in apartments during childhood more frequently than nonatopic controls (53.6% vs. 38%, p = 0.04). Atopic group members, particularly the asthmatics, had lower sibship size (≤ 1 sibling) (41.3% vs. 16.7%, p = 0.0003) and nonatopic subjects had higher sibship size (≥3 siblings) (73.8% vs. 43.4%, p<0.0001). The rate of maternal atopy was higher in the atopic group (24.7% vs. 9.5%, p = 0.02). Place of birth, sibship size, and atopic status of the mother, but not house type, were found to be significant in the multivariate analysis. As a conclusion, rural life-style seems to have a protective effect on the development of atopy in Turkey. On the contrary, factors directly or indirectly related to urban life-style during early childhood were found to be associated with the allergic airway diseases.

Increased Serum IgE in Alcoholics: Relationship With Th1/Th2 Cytokine Production by Stimulated Blood Mononuclear Cells

Background: Increased serum immunoglobulin (Ig) E values are frequently found in alcoholics. Cytokines produced by T-helper-2 (Th2) lymphocytes are required for IgE synthesis. Chronic alcoholism is associated with altered cytokine balance. This study analyzed the relationship between Th1 and Th2 cytokine production by stimulated peripheral blood mononuclear cells (PBMCs) and serum IgE levels, both in atopic and nonatopic alcoholics. Methods: Twenty-five patients admitted to the hospital with alcohol withdrawal syndrome were included in the study. Five were classified as atopic and 20 as nonatopic by means of skin-prick tests. Interleukin-4 (IL-4), IL-10, IL-12, IL-13, and interferon γ were measured in the supernatants of 48-hr cultures of PBMCs stimulated with phytohemagglutinin. Total serum IgE was measured by chemiluminescent enzyme immunoassay. Results were compared with those of 15 healthy controls (seven atopics and eight nonatopics). Results: Total serum IgE concentrations were higher in alcoholics than in controls, in both atopic and nonatopic subjects. The ratio of IL-4 to interferon γ production by phytohemagglutinin-stimulated PBMCs (as an approach to Th2/Th1 balance) was significantly lower in alcoholics than in healthy controls, both in the atopic and in the nonatopic group. No difference was observed regarding IL-10, IL-12, and IL-13 production between alcoholics and controls. No correlation was demonstrated between cytokine production and total serum IgE levels in any group. Conclusions: Increased total serum IgE is observed in alcoholics together with a paradoxically low ratio of Th2 to Th1 cytokine production by phytohemagglutinin-stimulated PBMCs. These findings are independent of the atopic status of patients.