F.20. Characterising T-Cell Reponses in Wasp Allergic Patients Receiving Immunotherapy

Abstracts respiratory virus, bacterial and fungal infectious diseases (up to 14 episodes per year) that decreases quality of life. Seasonal manifestations of pollen allergy are also longer and more severe comparing with patients of same age group and same spectrum of sensitization but without secondary immunodeficiency. Our aim was to evaluate clinical manifestations, and deviations in immune system, and try to influence on them during Specific Immunotherapy (SIT). 100 patients were randomized into experimental and control groups and were undergoing clinical, allergological, immunological examinations. We used skin prick tests, provocation tests, microscopy of nasal smears. There were considerable changes in immune status comparing with non allergic humans but the most impressive was clinical information in these patients that was obtained by anamnestic data. Comparing with controls in most of them the spectrum of allergic sensitization increased each year (they were no specific treatment before), and number of shock organs increased as well. We used intranasal mucosal specific immunotherapy, as the most effective scheme of specific therapy in all patients with pollen allergy. We implemented our theory in patients with PA and SID by adding to traditional scheme of SIT immunomodulation by highly-molecular polymers in rectal suppositories (6 mg in a day 20 days) and revealed positive results. All positive trends were more intensively expressed in the experimental group and faced in clinical efficacy, positive changes in immune status, decreasing of amount of total IgE, specific IgE increasing of Th1 and decreasing of range of allergic and infectious exacerbations. doi:10.1016/j.clim.2006.04.057

F.18. ‘‘In Vitro’’ and ‘‘in Vivo’’ Immunomodulant Activity of PSMIX (Lactobacillus Paracasei I 1688 and Lactobacillus Salivarius I 1794). Annamaria Castellazzi,1 Maria antonietta avanzini,2 Mara Oliveri,1 Chiara Valsecchi,1 Patrizia Malfa,3 Gianluigi Marseglia. 1Pediatric Sciences, University of Pavia, Pavia, Italy; 2Laboratori sperimentali, IRCCS Policlinico San Matteo, Pavia, Italy; 3Laboratory, Progefarm, Novara, Italy. Many studies showed the role of lactic acid producing bacteria(LAB)in promoting the human and animal health. Some LAB strains can influence the immune response against pathogenic agents and tumoral degenerations and are able to increase the immune response at mucosal and systemic levels.The aim of the present work was to evaluate the ‘‘in vitro’’ action of the Lactobacillus paracasei I 1688, Lactobacillus salivarius I 1794 and of a commercial mix of them (PSMIX, Proge Farm) on lymphocytes of adult healty individuals.Moreover, to confirm the probiotic action, we studied , in 20 atopic children, NK activity, plasma cytokine profile and subpopulations of lymphocytes before and after 30 days ingestion of the same probiotics. The results show that the lymphocytes of healthy adult subjects respond with proliferation to the presence of probiotic bacteria in culture significantly less than in response to Candida albicans.The percentage of T-cells more implicated in the proliferative response has regulatory morphology (CD4+CD25+) and the

S57 cytokine production is of Th1 profile (TNF-alpha NIFN-gamma NIL-12).The ‘‘in vitro’’ production of IL-10, a cytokine involved in immunomodulation, is more significantly increased after stimulation with probiotic bacteria than with Candida albicans. In ‘‘in vivo’’ studies, in the atopic population we could confirm the same results. These data can support the idea that the LAB examinated, mainly in the proportion present in the mix, can be useful to modulate the immune response in allergic disease from a TH2 to a TH1 profile. doi:10.1016/j.clim.2006.04.058

F.19. Rhinitis Is a Risk Factor for Skin Reactivity to Aeroallergens in Asthma. Fardin Eghtedari, Sara Kashef, Mohammad Amin Kashef. Allergy Research Center, Shiraz University of Medical Sciences, Shiraz, Iran. Background: Asthma is one of the most common chronic diseases among children and adults. Prevalence of asthma has been increased for the last thirty years. It has been hypothesized that atopy is a strong background predisposing to asthma. The aim of this study was to determine whether rhinitis is significantly associated with skin reactivity in asthma patients. Methods: Skin prick testing (SPT) was performed on 85 asthmatic cases aged between 1 and 49 years (mean 12.9 years) during a period of two years (20032005). The reaction was considered to be positive if the mean wheal diameter was at least 3 mm greater than that of the negative control. Results: 47 (55%) patients had at least one positive SPT to pollen (33 cases), mite (20 cases) or mould (5 cases). Concurrence of rhinitis was detected in 24 cases including 18 patients with positive SPT. Skin reactivity was significantly associated with rhinitis in this cohort of patients (P = 0.022) Conclusion: Rhinitis is associated with skin reactivity in asthma patients. It could be an emphasis on atopy as a strong background predisposing to asthma. Keywords: asthma, atopy, rhinitis. doi:10.1016/j.clim.2006.04.059

F.20. Characterising T-Cell Reponses in Wasp Allergic Patients Receiving Immunotherapy. Aamir Aslam, David Warrell, Graham Ogg. John Radcliffe Hospital, Oxford, United Kingdom. Wasp allergy can be treated with subcutaneous injections of increasing doses of wasp venom. We are testing the hypothesis that this acquisition of tolerance is associated with changes in the phenotype of wasp venom specific T-cells. By using the Elispot assay, we have shown that PBMC from wasp allergic patients (n = 12) have a higher frequency of IL-4 secreting T-cells than controls (n = 9) in response to whole wasp venom: 170 spot forming units (sfu)/million PBMC and 0.7 sfu/million PBMC respectively, p b 0.05. Furthermore, this response is largely mediated by CD4+ T-cells. We have confirmed that Ves V1, Ves V2 and Ves V5 are the components of wasp venom that are responsible for these IL-4 responses by separating wasp venom by size-exclusion and ion-

S58 exchange chromatography and SDS-PAGE and subsequently identifying the dominant reactive proteins by tandem mass spectrometry. Using Ves V1 and Ves V5 derived peptides we have shown the presence of ex-vivo IL-4, interferon gamma and IL-10 responses in wasp allergic patients. Additonally, we have begun to map individual epitope responses and characterise their HLA restriction. By elucidating the mechanisms involved in induction of clinical tolerance in humans, we hope to identify novel therapeutic strategies in individuals with allergic disease and other forms of hypersensitivity. doi:10.1016/j.clim.2006.04.060

F.21. Atopy As a Cause of Delayed Maturation of the Immune System. Isaac Melamed, Angela McDonald, Ben Jessen. -, 1st Health Centers, Centennial, CO. Introduction: Recurrent infections, mostly ear or respiratory in nature, can be the first presentation in children with atopy. Unknown is the linkage of the immune system to atopy. Transient hypogammaglobulinemia (TH) is characterized by a delayed maturation of the immune system. The pathogenesis of the delay is unknown. In this study, we speculate that atopy may play a role in this delayed maturation. Clinical Data: Based on our clinical observation, we followed a group of 15 children (aged 15 to 48 months) who had been diagnosed with transient hypogammaglobulinemia (IgG: 130-270 mg/dl), who had no response to pneumococcal Ab and polio and who had reduced response to isohaemaglutinines. Their infectious presentations included the following infections: ear (12), respiratory (9) and skin (3). Results: Our follow-up, conducted three-five years after the initial diagnosis of hypogammaglobulinemia, revealed that the children’s IgG levels had returned to normal limits and that 12 out of 15 of the children had developed full clinical and laboratory pictures of atopy-related diseases. All 12 atopic children had positive skin tests to environmental allergens and 9 of them had high IgE and positive RAST tests. Their clinical presentation: 4 with eczema, 9 with allergic rhinitis and 6 with a combination of allergic rhinitis and reactive airway disease. Conclusion 1. Recurrent ear infections and upper respiratory infections are the preliminary presentations for atopy in children. 2. A high percentage of children with transient hypogammaglobulinemia will develop atopic-related diseases. 3. Atopy may be involved in delayed maturation of the immune system and the presentation of transient hypogammaglobulinemia. doi:10.1016/j.clim.2006.04.061

F.22. Efficacy of Immunomodulating Therapy in Children with Severe Asthma. Elena Urban, Michael Rudenko. Research Institute of Clinical Immunology and Allergology, Rostov State Medical University, Rostov-on-Don, Russian Federation. Inhaled glucocorticosteroids are used in treatment of severe asthma as basic therapy. Such long therapy in children

Abstracts can lead to inevitable changes in immune system. To increase efficacy of treatment and correction of immune deviations it is possible to add immunomodulating therapy. The aim of this study was to investigate the efficacy of highly molecular polymers in children with severe bronchial asthma. This drug has distinct immunomodulation, detoxication functions and antiphlogistic effects. We examined 40 children 10 -14 years old with bronchial asthma. 1 group: 20 patients got daily 250 microgram of glucocorticosteroid and highly molecular polymers 6 mg in rectal suppositorium twice a week - 6 weeks. And control group 20 persons got daily 250 microgram glucocorticosteroid. During that period we estimated the dynamics of clinical symptoms, peakflowmetry, curve flow - forced inspiration volume with bronco-dilatation test for reversibility and daily necessity in h2-agonists. Immunological examination was used before and after the treatment CD3+, CD4+, CD8+, CD16+, CD20+, CD95+, HLA-DR, IgM, IgG, IgA, IgE, CIC, functional activity of phagocytes. In the result of treatment frequency and intensity of exacerbations in 1 group decreased to 25-35%, necessity in h2-agonicts decreased in 1, 2 1, 4 times. Increasing of forced inspiration volume and life capacity of lungs from 10% to 20%, normalization of immune regulating index (1, 9 F 0, 2), versus control (1, 3 F 0, 1), decreasing of total IgE (96 F 0, 1), versus control (119+0, 15), increasing of IgA (1, 7 F 0, 2), versus control (0, 9 F 0, 15), decreasing of CIC (83 F 0, 3), versus control (105 F 0, 25), increasing of phagocyte activity (1, 6 F 0, 2), versus control (1, 2 F 0, 4). Thus we can summarize that adding of highly molecular polymers lead to positive dynamics in clinical and laboratory rates in children with severe bronchial asthma. doi:10.1016/j.clim.2006.04.062

F.23. Polymorphisms of Toll-Like Receptor 2 and 4 Genes in Asthma and COPD. Nancy Larocca,1 Juan DeSanctis,2 Felix Toro,3 Dolores Moreno.4 1Biologia Molecular, Instituto de Inmunologia, Caracas, Venezuela; 2Biologia Molecular, Instituto de Inmunologia, Caracas, Venezuela; 3Biologia Molecular, Instituto de Inmunologia, Caracas, Venezuela; 4 Neumonologia, Hospital Clinico Universitario, Caracas, Venezuela. Human Toll-like receptor family (TLRs) play a central role in the initiation of cellular innate immune response. Individual TLRs recognize distinct pathogen associated molecular patterns in specific classes of microbes. Binding of this pathogen associated molecular patterns to TLRs triggers a complicated series of events leading to increased expression of proinflamatory genes. Studies of people with specific polymorphisms in genes encoding TLRs can elucited relationships between TLR signalig and human disease. We investigated, by using a polymerase chain reaction restriction fragment length polymorphism method, the possible association between the polymorphisms of TLR2 (Arg677Trp and Arg753Gln) and TLR4 (Asp299Gly and Thr399Ile)genes with suceptibility or severity of Asthma and COPD. Our study population consisted of 100 patients with Asthma, 100 patients with COPD and a control group of 100 healthy individuals. Not found TLR2 polymorphisms in the patients o controls. We found