Novel strategies in immunotherapy for allergic diseases.

Conventional immunotherapy (IT) for optimal control of respiratory and food allergies has been fraught with concerns of efficacy, safety, and tolerability. The development of adjuvants to conventional IT has potentially increased the effectiveness and safety of allergen IT, which may translate into improved clinical outcomes and sustained unresponsiveness even after cessation of therapy. Novel strategies incorporating the successful use of adjuvants such as allergoids, immunostimulatory DNA sequences, monoclonal antibodies, carriers, recombinant proteins, and probiotics have now been described in clinical and murine studies. Future approaches may include fungal compounds, parasitic molecules, vitamin D, and traditional Chinese herbs. More robust comparative clinical trials are needed to evaluate the safety, clinical efficacy, and cost effectiveness of various adjuvants in order to determine ideal candidates in disease-specific and allergen-specific models. Other suggested approaches to further optimize outcomes of IT include early introduction of IT during an optimal window period. Alternative routes of administration of IT to optimize delivery and yet minimize potential side effects require further evaluation for safety and efficacy before they can be recommended.

Tobacco smoke exposure and respiratory morbidity in young children.

OBJECTIVE: Secondhand smoke exposure is a potentially preventable cause of significant respiratory morbidity in young children. Our study aimed to quantify respiratory morbidity in young children exposed to secondhand smoke to identify potentially modifiable factors. MATERIALS AND METHODS: This study was embedded in a prospective birth cohort study of pregnant women and their children from fetal life onwards in Singapore (Growing Up in Singapore Towards healthy Outcomes, or GUSTO). Data on prenatal, antenatal and postnatal active and secondhand tobacco smoke exposure were obtained by an investigator-administered questionnaire for the periods before pregnancy, at 26-28 weeks' gestation and 24 months after delivery. Data on respiratory morbidity (wheezing episodes, croupy cough, nebuliser use, snoring) and other morbidity (fever, hospitalisation, ear infection) of the child was collected at week 3 and at months 3, 6, 9, 12, 15, 18 and 24 after delivery. Information on parental atopy and potential confounders such as socioeconomic status and maternal educational level were also obtained. Statistical analysis of the data was performed to quantify any significant differences in incidence of respiratory morbidity in children exposed to tobacco smoke in utero and postdelivery, compared with those in smoke-free environments. RESULTS: Women who smoked regularly prior to pregnancy comprised 12.5% (n=155) of the study population; this number fell to 2.3% (n=29) during pregnancy. Mothers exposed to secondhand smoke in the household before pregnancy comprised 35.7% of the study population (n=441) and 31.5% (n=389) were exposed during pregnancy. Postnatally, the prevalence of secondhand tobacco smoke exposure from birth to 2 years of age was 29% (n=359). Participants of Malay ethnicity (p<0.001), mothers with no or primary level education (p<0.001) and mothers with low socioeconomic status (p<0.001) had the highest exposure to tobacco smoke. Offspring secondhand smoke exposure at home by 12 months and by 24 months of age was associated with an increase in hospital admissions due to respiratory disease (RR 1.89, 95% CI 1.02 to 3.50, p=0.04 by 12 months and RR 1.64, 95% CI 1.05 to 2.55, p=0.03 by 24 months) as well as all-cause hospitalisation (RR 1.57, 95% CI 1.14 to 2.17, p=0.01 by 12 months and RR 1.49, 95% CI 1.17 to 1.90, p=0.001 by 24 months), adjusting for parental atopy and child atopic dermatitis. Participants exposed to secondhand smoke by 12 months postdelivery had a significantly increased risk of having at least one wheezing episode (RR 1.71, 95% CI 1.38 to 2.11, p<0.001). CONCLUSIONS: Secondhand smoke exposure during the prenatal and postnatal periods is associated with increased respiratory morbidity in children. Opportunistic screening and targeted smoking cessation counselling for parents at child hospital admissions and well-child outpatient visits, as well as preconception smoking cessation counselling for future pregnancies, may be beneficial to protect the child from negative health impacts.

Clinical and immunochemical profiles of food challenge proven or anaphylactic shrimp allergy in tropical Singapore.

BACKGROUND: Shellfish allergy in Singapore is highly prevalent, and shrimp allergy is the most common. OBJECTIVE: This study aims to evaluate the clinical characteristics and immunological phenotype of shellfish allergy in this population. METHODS: Patients with self-reported shellfish allergy were recruited from outpatient clinics of three large hospitals and from a population survey. Open oral food challenges (OFC) to glass prawn (Litopenaeus vannamei) and tiger prawn (Penaeus monodon) were carried out on all patients except for those who had a history of severe anaphylaxis. Skin prick tests (SPT) and specific IgE to crude and recombinant allergens were carried out to evaluate shrimp and dust mite sensitization. Immunoblots were used to assess IgE-binding proteins. RESULTS: The 104 patients recruited were categorized into shellfish allergic (SA) when OFC was positive or had a history of severe anaphylaxis (n = 39), shellfish tolerant (ST) when OFC was negative (n = 27), and house dust mite positive controls (HDM(+) ) who were ST (n = 38). Oral symptoms (87.1%) were the predominant clinical manifestation. Positive challenge doses ranged from 2 to 80 g of cooked shrimp, with 25/52 patients reacting to either one or both shrimps challenged. The presence of specific IgE to shrimp either by SPT and/or ImmunoCAP(®) assay provided diagnostic test sensitivity of 82% and specificity of 22.2%. The inclusion of specific IgE to shrimp tropomyosin and IgE immunoblots with shrimp extracts did not improve the diagnostic proficiency substantially. CONCLUSIONS AND CLINICAL RELEVANCE: This study highlights the predominance of oral symptoms in shrimp allergy in tropical Asia and that a high provocation dose may be necessary to reveal shrimp allergy. Furthermore, specific IgE diagnostic tests and immunoblots were of limited use in this population.

International consensus on (ICON) pediatric asthma.

Asthma is the most common chronic lower respiratory disease in childhood throughout the world. Several guidelines and/or consensus documents are available to support medical decisions on pediatric asthma. Although there is no doubt that the use of common systematic approaches for management can considerably improve outcomes, dissemination and implementation of these are still major challenges. Consequently, the International Collaboration in Asthma, Allergy and Immunology (iCAALL), recently formed by the EAACI, AAAAI, ACAAI, and WAO, has decided to propose an International Consensus on (ICON) Pediatric Asthma. The purpose of this document is to highlight the key messages that are common to many of the existing guidelines, while critically reviewing and commenting on any differences, thus providing a concise reference. The principles of pediatric asthma management are generally accepted. Overall, the treatment goal is disease control. To achieve this, patients and their parents should be educated to optimally manage the disease, in collaboration with healthcare professionals. Identification and avoidance of triggers is also of significant importance. Assessment and monitoring should be performed regularly to re-evaluate and fine-tune treatment. Pharmacotherapy is the cornerstone of treatment. The optimal use of medication can, in most cases, help patients control symptoms and reduce the risk for future morbidity. The management of exacerbations is a major consideration, independent of chronic treatment. There is a trend toward considering phenotype-specific treatment choices; however, this goal has not yet been achieved.

Filaggrin mutations are associated with recurrent skin infection in Singaporean Chinese patients with atopic dermatitis.

BACKGROUND: Loss-of-function (null) mutations within the filaggrin (FLG) gene are a strong risk factor for atopic dermatitis (AD). We hypothesized that the absence or reduction of the filaggrin protein could compromise skin barrier and increase patients' susceptibility to recurrent skin infection. OBJECTIVES: To investigate the association between FLG-null mutations and the risk of recurrent skin infection among a series of patients with AD in Singapore. METHODS: This study included 228 Singaporean Chinese patients with AD with at least 1year of follow-up at the time of recruitment between January 2008 and December 2009 at the National Skin Centre in Singapore. Each patient had their medical records reviewed for history of skin infection in the preceding year and was genotyped for 22 FLG-null mutations. RESULTS: Compared with those without the FLG-null mutations, patients with AD who had FLG mutation(s) had approximately a seven times increased risk of more than four episodes of skin infection requiring antibiotics in the past year (odds ratio 6·74; 95% confidence interval 2·29-19·79). This risk was much greater in those with mild or moderate disease, and was present in both users and nonusers of oral steroids. CONCLUSION: This study highlights a novel association between FLG-null mutations and an increased susceptibility to recurrent bacterial skin infection among patients with AD.

Wide spectrum of filaggrin-null mutations in atopic dermatitis highlights differences between Singaporean Chinese and European populations.

BACKGROUND: Null mutations in the filaggrin gene (FLG) cause ichthyosis vulgaris (IV) and predispose to atopic dermatitis (AD). Cohort studies in Europe and Japan have reported an FLG mutation carrier frequency of between 14% and 56%, but the prevalent European FLG mutations are rare or absent in Chinese patients with IV and AD. OBJECTIVES: To investigate further the spectrum of FLG-null mutations in Chinese patients and to compare it with that in other populations. METHODS: We conducted comprehensive FLG genetic analysis in a discovery cohort of 92 Singaporean Chinese individuals with IV and/or moderate-to-severe AD. All detected FLG mutations were then screened in a cohort of 425 patients with AD and 440 normal controls. Results In total, 22 FLG-null mutations, of which 14 are novel, were identified in this study; the combined null FLG genotype of 17 mutations detected in cases and controls showed strong association with AD [Fisher's exact test; P = 5·3 × 10⁻9; odds ratio (OR) 3·3], palmar hyperlinearity (Fisher's exact test; P = 9·0 × 10⁻¹5; OR 5·8), keratosis pilaris (Fisher's exact test; P = 0·001; OR 4·7) and with increased severity of AD (permutation test; P = 0·0063). CONCLUSIONS: This study emphasizes the wider genetic landscape of FLG-null mutations in Asia that is slowly emerging.

Monitoring and treatment practices of childhood asthma in Singapore: a questionnaire study.

INTRODUCTION: Asthma is the most common chronic disease in children in Singapore. More than 20 percent of children will have been diagnosed with asthma by the age of 15 years. Most children are seen in the primary care setting, thus it is of value to study the management practices, especially of general practitioners, with comparison to gold standards. The aims of the study were to investigate: (a) Methods of monitoring asthma control; (b) Practices in managing acute exacerbations; and (c) Choice of therapy in maintenance treatment. METHODS: 2,100 questionnaires consisting of 35 questions were sent by post to general practitioners and various paediatric doctors throughout Singapore. 173 valid responses were received and results were compared to the 2006 Global Initiative for Asthma guidelines. RESULTS: 76.3 percent of respondents were general practitioners. 89.1 percent did not use symptom score cards / diaries. 37.6 percent did not use peak-flow meters / spirometers. 83.8 percent used a short-acting beta-agonist in acute exacerbations, but only 41.0 percent used oral corticosteroids in outpatients. A significant number used long-acting beta-agonists (LABA) in combination with inhaled steroids (29.5-41.6 percent) or as monotherapy (5.8-8.7 percent) for maintenance treatment. 91.3 percent never used immunotherapy in practice. CONCLUSION: Greater usage of diaries / score cards can be encouraged along with objective peak flow / spirometry measurements. Management of acute exacerbations is appropriate but corticosteroids are under-prescribed by most doctors. LABA continues to be prescribed for maintenance despite a lack of established safety profile for infants, along with recommendations that they only be used selectively in patients poorly-controlled by medium-dosage inhaled corticosteroids.

Prevalence of drug allergy in Singaporean children.

INTRODUCTION: Adverse drug reactions (ADRs) are a common medical problem in children, affecting up to 15 percent of children, according to the literature. However, most studies on ADRs were performed in a hospital setting, and studies in the general population are limited. The current study aims to estimate the prevalence of ADRs in a large number of non-selected Singaporean children. METHODS: School children, aged 7-16 years, from 25 random schools were screened via a self-reported questionnaire on ADRs, and parents of the selected children were then followed up with a telephone interview to obtain additional information on specific manifestations, diagnosis and allergy testing. RESULTS: The prevalence of an ADR in children was 5.4 percent, with 56.7 percent of cases reporting an ADR to beta-lactam antibiotics. Dermal manifestations were reported in 60 percent of all ADRs, while multiple drug allergies accounted only for 3.8 percent. Only 6.9 percent of the children who experienced an ADR were referred to a hospital for further investigations. CONCLUSIONS: ADRs were associated with a positive history of atopy, increased income level and Chinese and Indian ethnicity, but not with gender or age. It is striking that most children suffering from a clinical ADR were not investigated further or referred for diagnostic tests. Many parents were unaware of the availability of drug allergy tests and feared compromising their children's health. This certainly could attribute to the high incidence of the over-reporting of ADRs in the general population.

Identification of human metapneumovirus and Chlamydophila pneumoniae in children with asthma and wheeze in Singapore.

INTRODUCTION: The aim of our study was to determine if human metapneumovirus (hMPV) and Chlamydophila pneumoniae (CP) could be detected in Singaporean asthmatic children and wheezing infants during an acute asthma attack. METHODS: The study was performed on 30 older children (mean age 9.8 years) and 30 young children (mean age 1.3 years), who were admitted with an acute exacerbation of wheezing. Nasopharyngeal aspirates were collected and tested by polymerase chain reaction for CP, and for a panel of viruses (hMPV, respiratory syncytial virus, adenovirus, influenza virus types A and B, parainfluenza virus types 1 and 3, and rhinovirus). RESULTS: hMPV was isolated in eight out of 60 children (13.3 percent), while CP was isolated in two cases. Overall, 48/60 (80 percent) samples were positive for the presence of viruses. CONCLUSION: In most of the children admitted because of acute wheezing, a virus could be detected. hMPV was isolated for the first time in Singapore in children who were admitted with an acute asthma attack.

Distinct pattern of commensal gut microbiota in toddlers with eczema.

BACKGROUND: Recent studies have demonstrated differences in the composition of gut microbiota in infants with and without allergic diseases, particularly eczema. METHODS: A case-control study involving 21 toddlers (age 3.0 +/- 0.5 years) with and 28 age-matched toddlers without eczema was conducted. Four groups of aerobic gut microbiota were identified and quantitated in stool samples grown on selective media. Three groups of anaerobes were enumerated by fluorescent in situ hybridization followed by quantitative flow cytometry. We also performed molecular typing of lactic-acid-producing bacteria (LAB) and enterococcal isolates to facilitate detailed analysis at species level by bacterial 16S rDNA sequencing. RESULTS: Toddlers with eczema harbored significantly lower counts of Bifidobacterium [(median 0.14 (25th and 75th percentile: 0.04 and 0.47) vs. 0.71% (0.16, 1.79) of cells acquired, p = 0.003)] and Clostridium [(0.28 (0.09, 0.78) vs. 0.83% (0.35, 1.82) of cells acquired, p = 0.012)] but significantly higher counts of total LAB [7.3 (6.1, 8.5) vs. 5.7 (4.4, 7.3) log CFU/g, p = 0.006] in particular enterococci [6.3 (4.8, 7.4) vs. 5.0 (3.4, 6.4) log CFU/g, p = 0.018]. There was no significant correlation between eczema severity score and bifidobacterial counts. CONCLUSION: The results further confirm previous reports that the gut microecosystem differs between children with and without eczema and extend them beyond infancy.

Perinatal risk factors for sensitization, atopic dermatitis and wheezing during the first year of life (PIPO study).

OBJECTIVE: To evaluate the influence of perinatal environmental factors on early sensitization, atopic dermatitis and wheezing during the first year. METHODS: Information on pregnancy-related factors, parental atopic history, environmental factors and the clinical course of the infant until age one was gathered by questionnaires, as part of a prospective birth cohort study (Prospective study on the Influence of Perinatal factors on the Occurrence of asthma and allergies [PIPO-study]). Quantification of total and specific IgE was performed in 810 children and their parents. RESULTS: Early sensitization was found in 107/810 (13%) of the infants. Multiple regression analysis showed that specific IgE in fathers was a risk factor for early sensitization in their daughters (adjusted odds ratios (OR(adj)) 2.21 (95% confidence interval (CI) 1.10-4.49); P=0.03), whereas in boys, day care attendance was shown to be protective for early sensitization (OR(adj) 0.38 (95% CI 0.20-0.71); P=0.001). Atopic dermatitis occurred in 195/792 infants (25%). Specific IgE in the mother (OR(adj) 1.52 (95% CI 1.06-2.19); P=0.02) and in the infant (OR(adj) 4.20 (95% CI 2.63-6.68); P<0.001) were both risk factors for the occurence of atopic dermatitis, whereas postnatal exposure to cats was negatively associated with atopic dermatitis (OR(adj) 0.68 (0.47-0.97); P=0.03). Postnatal exposure to cigarette smoke (OR(adj) 3.31 (95% CI 1.79-6.09); P<0.001) and day care attendance (OR(adj) 1.96 (95% CI 1.18-3.23); P=0.009) were significantly associated with early wheezing, which occurred in 25% (197/795) of the infants. CONCLUSION: The effect of paternal sensitization and day care attendance on sensitization is gender dependent. Maternal sensitization predisposes for atopic dermatitis, whereas postnatal exposure to cats had a protective effect.

Anaphylaxis after hamster bites--identification of a novel allergen.

BACKGROUND: Hamsters are popular household pets and anaphylaxis after their bites have described. However, the putative allergen has not been identified. OBJECTIVE: This study was conducted to identify the allergen causing dwarf hamster (Phodopus sungoris) bite-induced anaphylaxis. METHODS: Two children with hamster bite-induced anaphylaxis were enrolled. They both had negative results to skin testing and specific IgE to hamster epithelium. However, they were both allergic to Dermatophagoides pteronyssinus (Der p). Identification of the putative IgE-binding allergens from the hamster saliva was performed using immunoblot analysis. RESULTS: A specific IgE-binding component at 21 kD in the hamster saliva was identified. ELISA inhibition tests showed partial inhibition with Der p. CONCLUSIONS: The putative allergen from the hamster saliva causing dwarf hamster-induced anaphylaxis was identified. Possible cross-reactivity with Der p was demonstrated. Further studies will be needed to identify the exact nature and function of this allergen.

Differences in circulating dendritic cell subtypes in cord blood and peripheral blood of healthy and allergic children.

BACKGROUND: Different types of circulating dendritic cells have been described. Dendritic cells influence differentiation of naive T lymphocytes into T helper type 1 (Th1) and Th2 effector cells. OBJECTIVE: The purpose of this study was to evaluate the number of circulating DC subtypes in peripheral blood of allergic and healthy children and in cord blood of neonates from allergic and non-allergic parents. METHODS: Circulating dendritic cells were flow cytometrically identified in whole blood samples as lineage (CD3, CD14, CD16, CD19, CD20, CD56) negative, CD34 negative and HLA-DR-positive cells. According to the expression of CD123 and CD11c, different DC subtypes were identified. RESULTS: Apart from DC1 (CD11c+ CD123dim+) and DC2 (CD11c- CD123high+), a third DC population was described with less differentiated phenotypic characteristics, namely CD11c- CD123dim+, and therefore defined here as less differentiated DC (ldDC). These ldDC represented the major DC population in cord blood and showed an age-depended decrease. The highest level of ldDC was detected in children with atopic dermatitis, whereas asthmatic children showed the lowest ldDC counts. Furthermore, high-dose inhaled corticosteroid treatment in asthmatic children was related to a decreased ldDC number. The number of circulating DC2 was significantly lower in allergic children, especially in asthmatics, compared to healthy children. In cord blood, no differences in DC subtypes were detectable between neonates at low and high risk for allergic disorders. CONCLUSION: These results indicate that, apart from DC1 and DC2, a third population of dendritic cells, identified as CD11c- CD123dim+ cells and defined as less differentiated DC, must be considered in the evaluation of circulating DC. Furthermore, DC2 counts were decreased in allergic children, especially in asthmatics, which might be the consequence of an increased recruitment to the target organs.

Critical evaluation of prognostic factors in childhood asthma.

Current knowledge of the natural history of asthma is improving through the establishment of a more precise definition of asthma linked with information from a number of large-scale longitudinal studies. Risk factors for the development of childhood asthma are now more clearly understood. They include gender, atopic status, genetic and familial factors, respiratory infections, and outdoor and indoor pollution (1). In the present review two types of asthma and their prognosis will be discussed: (1) Asthma in preschool children and its risk factors for evolution towards persistent childhood asthma. (2) Asthma in older children and its risk factors for evolution towards adult asthma.

Effect of salbutamol on lung function parameters of healthy children.

UNLABELLED: In the present study, the effect of 200 microg salbutamol compared to placebo was evaluated on lung function parameters of 37 healthy children aged 7-14 years. Salbutamol or placebo were administered, using a single blind study design, and spirometry was performed before and after 10 min of inhalation. At the time of the study, all children were symptom-free and had not suffered from any respiratory infection during the previous 4 weeks. The administration of salbutamol resulted in a significant increase of mean forced expiratory volume in 1 s (111%-115%, P<0.05), maximal expiratory flow at 50% of forced vital capacity (101%-110%, P<0.05) and maximal expiratory flow at 25 % of forced vital capacity (96%-115%, P<0.05). The administration of placebo resulted in no significant change in lung function parameters. CONCLUSION: The administration of 200 microg salbutamol results in the occurrence of a small but significant bronchodilation in healthy, non-asthmatic children.

The role of inflammation in the development of chronic lung disease in neonates.

UNLABELLED: Chronic lung disease (CLD) has been associated with chorioamnionitis and upper respiratory tract colonisation with Ureaplasma urealyticum. The aim of this review is to describe the increasing evidence that inflammation plays a critical role in the early stages of CLD of the neonate. Ongoing lung damage in the premature infant may be caused by failure to downregulate and control this inflammatory response. Tumour necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6) and IL-8 are important pro-inflammatory cytokines of which IL-8 is an important chemotactic factor in the lung. Data suggest that preterm newborns with lung inflammation may be unable to activate the anti-inflammatory cytokine IL-10. Therefore, early post-natal anti-inflammatory therapy could help in preventing development of CLD. Prophylactic dexamethasone therapy cannot yet be recommended. There are a number of potential interactions between surfactant and cytokine effects on the preterm lung which have not been evaluated. Surfactant protein A may be an important modulator of the immune response to lung injury. The role of high-frequency ventilation in the prevention of CLD still remains unclear. CONCLUSION: Many aspects of the pathogenesis of the inflammatory response in the development of chronic lung disease remain to be elucidated. Further research to identify preterm infants at highest risk for the development of this multifactorial and complex disease is needed.

Higher occurrence of asthma-related symptoms in an urban than a suburban area in adults, but not in children.

In young adults, a higher occurrence of asthma-related symptoms was found in an urban than an adjacent suburban area in a survey performed in 1991. The authors now wondered whether such differences could be established in other age groups. The present study (in 1996) included 14,299 subjects, aged 5-75 yrs, of a random sample of the general population in the same two adjacent areas: the centre of Antwerp (Belgium) and its south suburban border. The standardized European Community Respiratory Health Survey (ECRHS) and International Study of Asthma and Allergies in Childhood (ISAAC) questionnaires were used to assess the occurrence of asthma-related symptoms. Higher rates were confirmed in urban compared to suburban Antwerp in adults (20-75 yrs), but no such area differences were found in children (5-8 and 12-15 yrs). Adjustment for a number of recorded risk factors did not seem to affect the area differences in asthma-related symptoms. Comparing the survey results of 1991 and 1996 in 20-44 yr old adults, the findings suggest a slight increase in reported respiratory symptoms in both areas. A higher occurrence of asthma symptoms was observed in the urban than suburban area in adults, but not in children. This might be explained by a progressive effect of long-term exposure to the "urban environment". However, longitudinal studies are necessary to further clarify the factors accounting for these age-related area differences.

Does the use of antibiotics in early childhood increase the risk of asthma and allergic disease?

BACKGROUND: One of the mechanisms evoked to explain the increasing prevalences of asthma and allergy, in particular among children, is the 'Western lifestyle' or 'hygiene' hypothesis. As early childhood infections are assumed to hold a protective effect on the development of asthma and allergies, the use of antibiotics at that sensitive age may lead to an increased risk of asthma and allergy. OBJECTIVE: The aim of this study is to investigate the association between the use of antibiotics in the first year of life and the subsequent development of asthma and allergic disorders. METHODS: In a population-based sample of 7-and-8-year-old children questionnaire and skin prick test data were collected from 1206 and 675 subjects, respectively. Prevalence rates of asthma, allergic disorders and skin test positivity were compared between children with and without early life use of antibiotics, taking into account other possible risk factors including early respiratory infections. The effect of genetic predisposition was investigated by stratified analyses of children with and without parental hay fever. RESULTS: The use of antibiotics during the first year of life was significantly associated with asthma (OR = 1.7, 95% CI 1.0-3.1), hay fever (OR = 2.3, 95% CI 1.3-3.8) and eczema (OR = 1.3, 95% CI 1.0-1.8). No significant relationship was found with skin test positivity (OR = 1.1, 95% CI 0.7-1.7). After stratification for the presence of parental hay fever, children without parental hay fever did not show any significant associations between antibiotics use and asthma or allergy, whereas in children with parental hay fever the use of antibiotics was significantly related with asthma (OR = 2.3, 95% CI 1.1-5.1), hay fever (OR = 2.8, 95% CI 1.5-5.1) and eczema (OR = 1.6, 95% CI 1.0-2.6), and of borderline statistical significance with skin test positivity (OR = 1.6, 95% CI 0.9-3.0). CONCLUSION: Early childhood use of antibiotics is associated with an increased risk of developing asthma and allergic disorders in children who are predisposed to atopic immune responses. These findings support recent immunological understanding of the maturation of the immune system.

Determination of T-cell subpopulations and intracellular cytokine production (interleukin-2, interleukin-4, and interferon-gamma) by cord blood T-lymphocytes of neonates from atopic and non-atopic parents.

This report describes the results of a prospective study on immunological markers in cord blood for the prediction of allergic diseases in children. First we evaluated methodological aspects of the flow cytometric technique on cord blood cytokine measurements. Subsequently, the T-cell subsets and percentage of cytokine-producing cord blood T-helper (Th) and T-suppressor/cytotoxic lymphocytes of neonates from atopic and non-atopic parents were compared. A group of 33 healthy, full-term newborn infants of whom 23/33 were at risk for atopy (i.e. having at least one parent with one or more atopic symptoms and positive specific immunoglobulin E [IgE] to at least one common inhalant allergen) was studied. A flow cytometric technique was used to analyze cord blood T-cell subsets and to determine the percentage of interleukin (IL)-2-, IL-4-, and interferon-gamma (IFN-gamma)-producing cord blood Th and T-suppressor/cytotoxic lymphocytes following stimulation with phorbol 12-myristate 13-acetate (PMA) and ionomycin. The percentage of CD3 (T lymphocytes), CD3+ CD4+ (Th lymphocytes), CD3+ CD8+ (T-suppressor/cytotoxic lymphocytes), CD19+ (B lymphocytes), CD3+ CD4+ CD45RO+ (memory Th lymphocytes), and CD3+ CD4+ CD45RA+ (naive Th lymphocytes) cells was unrelated to parental atopic status. PMA stimulation augmented the percentage of IL-2- and IFN-gamma-producing Th and T-suppressor/cytotoxic lymphocytes, whereas the number of IL-4-producing T lymphocytes remained very low or undetectable. No differences in the percentage of TL-2-, IL-4- and IFN-gamma-producing Th and T-suppressor/cytotoxic lymphocytes were found between neonates from atopic and non-atopic parents. These results will be re-evaluated when the atopic status of the children at the age of 1 and 2 years can be assessed.