11^th International Conference on Allergy, Asthma & Clinical Immunology September 07-08, 2017 Edinburgh, Scotland

Barbara De Servi has obtained a Biology degree at the University of Milan in 1998 and in 2002 the PhD in physiopathology at University of Milan and at DKFZ, in Heidelberg. In 2005-2006, she was post-doctoral fellow at the University of Verona medical school. Since 2007, she is study director at VitroScreen, in charge to develop tailor-made "omics" research models and identify toxicity pathways on 3D human tissue models.

Statement of the Problem: Nasal irrigation with saline solutions is frequently used for relief of rhinitis symptoms. Literature suggests that manganese (Mn) contributes to a decrease in allergic nasal response. In vitro research studies were conducted on 3D model of airway epithelium to evaluate efficacy and safety of a Mn-enriched seawater solution called Stérimar Allergic Nose (SAN). Methodology: The 3D Reconstituted Human Nasal Epithelium model (RHNE) was treated with 10 μL of SAN twice a day for four days to simulate repeated use (full strength) or untreated (control). For epithelium integrity (safety), the control and SAN-treated cultures were analyzed for: Trans-Epithelial-Electrical-Resistance (TEER) on days 1 (D1) and 4 (D4) post-treatment, and release of Lactate Dehydrogenase (LDH) and Interleukin 8 (IL-8) daily from D1 to D4. For efficacy, Mucociliary Clearance (MCC) and stimulation of epithelium-regeneration were assessed. MCC was measured by video-microscopy on D1 and D4 after the same treatment regimen. For epithelium-regeneration, RHNE was treated with 30 μL of SAN or saline. After a glass capillary injury, made 1 hour after treatment, regeneration stimulation was assessed as a percentage of wound closure by comparative photography immediately after the injury and 2, 6, 22 and 30 hours later. Findings: SAN showed an average TEER of 302 and 323 ohm.cm2 (p<0,001) on D1 and D4, respectively, safely above tissue integrity limit (100 ohm.cm2). LDH and IL-8 releases were similar for SAN and control at all-time points, also confirming epithelium integrity and safety. SAN showed a significant MCC increase as compared to control (P<0.01). Furthermore, SAN showed faster and greater wound closure than control (86.59% for SAN versus 50.65% at 22 hours). Conclusion: SAN demonstrated efficacy and safety in the in vitro assays. The results support the use of Mn-enriched SAN in relief of rhinitis symptoms.

Gilbert Glady has completed his MD at the age of 27 years from Strasbourg university of medicine and postdoctoral studies from Besançon and Paris-Nord universities of medicine. He got during all these years an expertise in immunology and immunogenetics and developed also interest for alternative medicines. So he becames at 2010 the creator of the BI(G)MED method and director of EBMA, the European association for training the medical profession at the BI(G)MED. He has participated in numerous international congresses in the field of immuno-allergology, infectiology and oncology with posters and oral presentations.

Inflammatory upper airway diseases, particularly chronic rhinosinusitis (CRS) and allergic rhinitis (AR), have a high worldwide prevalence. CRS and AR involve sustained and exaggerated inflammation that is associated with marked changes in gene and protein expression under tight regulation. MicroRNAs represent one of the fundamental epigenetic regulatory mechanisms used by cells that can mediate posttranscriptional gene silencing of target genes. As fine tuning regulators of gene expression, miRNAs are involved in diverse biologic processes, including cell proliferation, apoptosis, and differentiation, organ development, metabolism, stress responses, and signal transduction. Emerging evidence implicates an involvement of miRNAs in shaping the inflammation pattern in upper airways. Studies regarding the roles of miRNAs in allergic diseases have multiplied during the last 4 years, and the functions of miRNAs in the regulation and pathogenesis of these diseases are more and more better characterized. Recently, miRNAs have been shown to be detectable in cell-free body fluids such as serum and plasma samples. The circulating miRNAs are protected from blood RNAses either by existing in cell membrane-derived vesicles such as exosomes or by forming a complex with lipid-protein carriers such as high-density lipoprotein. So it becomes possible to use such kind of molecules for a therapeutic purpose, and that is what achieve the Bio Immun(G)en Medicine – BI(G)MED – by introducing high diluted microRNAs in nano compounds looking for a fine regulation in different upper airways diseases with an allergic etiology.

Tasuku Kawano completed his PhD at Tohoku Medical and Pharmaceutical University in 2009. He is presently working as an assistant professor in the Department of Pathophysiology at Tohoku Medical and Pharmaceutical University. He aims to reduce asthma patients. He studies elucidation of asthmatic onset mechanism based on nerve-endocrine-immunity axis. He has been trying for the creation of new fields for the development of novel medicines and preventive drugs in asthma. He has published “The involvement of central nervous system histamine receptors in psychological stress-induced exacerbation of allergic airway inflammation in mice” Allergol Int. 2016 Sep; 65 Suppl:S38-44

Statement of the Problem: Allergic asthma is characterized by Th2 type inflammation, essentially due to a breakdown of immune tolerance to an environmental allergen. Etiologically, experiences of early-life stress have been demonstrated to be associated with heightened prevalence of adult asthma. However, mechanisms underlying the stress leading to the development of asthma are poorly understood. Therefore, we examine if early-life stress increases the susceptibility to adult-onset asthma through the inhibition of the development of the respiratory tolerance. Methodology & Theoretical Orientation: Female BALB/c pups were sensitized by intraperitoneal injections of ovalbumin (OVA)/Al(OH)3 on postnatal days (PND) 24 and 29. Respiratory tolerance was induced by the inhalation of OVA on PNDs 18 and 21 before the sensitization. Maternal separation (MS) was used as a model of early-life stress and repeated from PND 17 to 22. On PND 76, the mice were challenged with OVA aerosol. Airway inflammation was evaluated with numbers of inflammatory cells in bronchoalveolar lavage fluid (BALF). The contents of IFN-, IL-4, IL-5 and IL-13 in BALF were measured by ELISA. The airway hyper-responsiveness (AHR) was assessed by methacholine-induced airflow obstruction. Conclusion & Significance: In tolerized mice, the numbers of inflammatory cells, the cytokine contents and AHR were remarkably decreased compared with those in non-tolerized mice. However, these effects of the tolerance were significantly reduced by MS exposure. These results suggested that early-life stress exposure has a potential to increase the risk of adult-onset asthma through the inhibition of the development of immune tolerance.

Heewon Lee is presently persuing her MS-Ph.D integrated course in Pharmaceutical Sciences College of Pharmacy, Ewha Woman’s University, Seoul, Korea. Completed B.S. in Pharmacy College of Pharmacy, Ewha Woman’s University, Seoul, Korea. Awarded with University Scholarship & also has a Korean Pharmacist License. Research includes studying the function of TCTP/HRF, a multifunctional protein. TCTP/HRF was found in body fluids of allergic patients and its importance was recognized, but the receptor and its mechanism of action were not yet known. Her ultimate goal was to find the HRF receptor, but attempts to isolate and identify it were unsuccessful. In the process, however, she discovered a functional domain that is thought to be the receptor binding domain of HRF. Antibodies targeting this region are being produced and the antibodies with good efficacy will be selected as HRF inhibitors.

We first reported that translationally controlled tumor protein (TCTP) acts as a histamine-releasing factor (HRF) associated with chronic allergic disease when it forms dimers. Despite the lack of signal peptide, HRF was found in nasal lavages, skin blisters and bronchoalveolar lavage fluids in late phases of allergic reaction. In a preliminary study, we found a significant increase in serum HRF levels in patients with asthma and allergic rhinitis. These results have led us to investigate whether HRF can be a therapeutic target for allergic diseases. By screening a phage-displayed 7-mer peptide library, we identified one peptide that showed strong affinity for the dimer TCTP (dTCTP). The peptide named dTCTP-binding peptide 2 (dTBP2) blocked the action of HRF by inhibiting binding to the cell surface. Specifically, dTBP2 inhibited the release of IL-8, an inflammatory cytokine, by inhibiting dTCTP-induced NF-κB and MAPK from human bronchial epithelial cell line BEAS-2B. In addition, dTBP2 dose-dependently reduced the symptom score and eosinophil recruitment to the nasal mucosa in OVA-induced allergic rhinitis mouse model, suggesting that in vivo inflammation-mediated airway pathology was alleviated. In this study, we showed that inhibition of dTCTP could alleviate allergic pathology and showed that dTCTP could be a new drug target for chronic allergic diseases such as allergic rhinitis.

Boris Ferko is presetly working at EURRUS Biotech GmbH, Austria.

We tested the efficacy of a novel biogenic peptidoamine compound, glutarimide histamine (XC8) in mouse, rat and guinea pig asthma models. Sensitized animals underwent oral treatments for at least 10 consecutive days with titrated doses of XC8 or corticosteroid reference drugs. In mice, XC8 efficiently inhibited eosinophilic lung inflammation of acute asthma disease onset, suppressed mucus hypersecretion, antigen-specific serum IgE or IgG1 titres, and methacholine-induced airway hyperresponsiveness (AHR). In Sephadex-induced migration of eosinophils in a rat model XC8 decreased the content of eosinophils in bronchalveolar lavages (BAL) 2.6-6.4 times. In guinea pig models of asthma and antigen-induced bronchospasm, XC8 reduced the number of degranulated mast cells and basophils in the lung tissue and the degree of degranulation. Moreover, XC8 also reduces hyperactivity of the lungs and reduces mortality of the animals from anaphylactic reactions. Chronic toxicity studies in rats and dogs revealed an excellent safety profile of XC8. In vitro experiments indicated that the mode of XC8 action might be associated with the suppression of glutaminyl cyclase - an enzyme that converts the immature form of chemokines (CCL2, CCL7, CCL8, CCL13) into the mature form by the reaction of pyroglutamination, thus suppressing the chemokine-driven migration of eosinophils and other cells into the inflammation area and the degranulation of mast cells and basophils. Our data demonstrate that XC8 efficiently suppresses experimental allergic asthma and provide support for its use as a treatment for human allergic asthma.

Luiz Werber-Bandeira is the Head of Clinical and Experimental Immunology Unit - Santa Casa de Misericórdia do Rio de Janeiro, Brazil. He has a degree in Medicine; completed his Post-doctorate in Immuno-Genetics and; PhD in Medicine-Immunology-Dermatology at Federal University of Rio de Janeiro. He is also specialized in Clinical Immunology-Allergy at Federal University of Rio de Janeiro. He is reorganizer of the clinical and experimental immunology unit - Santa Casa da Misericórdia, Rio de Janeiro.

Background: The gene encoding the adrenergic β2 receptor (ADRβ2) located on chromosome 5q31-32 presents a high allelic diversity characterized by the presence of several single nucleotide polymorphisms (SNPs) associated with different receptor activities at the cellular levels. 45 SNPs have been identified along this gene, some with significant effect on clinical response to β2-agonists. The allelic distribution varies with ethnicity and due to alterations in sensitization and down regulation of these receptors. The existing studies refer to ethnically homogeneous populations, with no data on the distribution of these SNPs in ethnically mixed populations, although certain haplotypes appear to directly affect receptor function. Objective: The objective of this study was to determine the prevalence of previously described SNPs in the coding region of the ADRβ2 gene, as well as to identify new SNPs in a group of individuals living in Rio de Janeiro through sequencing technique. Patients & Methods: DNA samples from 162 individuals were subjected to PCR amplification and genotyping by sequencing. Results: Analysis of the sequences generated identified the presence of nine different SNPs 46 (G>A), 66 (C>T), 79 (C>G), 252 (G>A), 455 (T>G), 470 (T>G), 491 C>T, 523 (C>A) e 579 (C>T) within the sequenced region (899 bp), from which, three: 455 (T>G), 470 (T>G), and 579 (C>T) were not yet described, and the characterization of 14 different haplotypes. The SNPs in codons 27 and 164, two of the most studied, showed different frequencies from those observed in other populations. The SNP in codon 27 was: 8.1% versus 24% in Caucasian-Americans and versus 18.7% in African-Americans. The SNP in codon 164 was: 4.6% versus 1% in Caucasian-Americans and versus 2% African-Americans. The SNP at codon 16, Arg16Gly, showed similar frequency to the one found in the literature in Caucasians and Chinese ethnicity: 54.9% vs. 39%, and 42% and 51% respectively. Conclusion: The results of this study, although represent a partial mapping of the coding region of the ADRβ2 gene, demonstrated the diversity of this gene, of great clinical importance. The results represent the first data on the partial mapping of the gene ADRβ2 in an ethnically mixed population.

Tatiana Chioro is a Brazilian Physician with large experience in Cosmetic Dermatology. She is Member of Brazilian Society of Dermatology, American Academy of Dermatology and European Academy of Dermatology and Venereology. She is also an MSc candidate at Hospital do Servidor Público Estadual – IAMSPE – São Paulo – SP – Brazil

Hyaluronic acid (HA) is biocompatible, easy-to-use, and reversible filler which is broadly-based filler in cosmetic medicine. More and more, non-expert physicians and non-physicians practitioners have been performing fillers in a large number of patients, which notices a high risk of unwanted outcomes, either in efficacy and safety fields. Unwanted results can mean overcorrection and asymmetries, as well as adverse events against these injectable fillers. Although HA-based fillers are defined as temporary materials, they can last up to 12 months or longer. Hyaluronidase is an endogenous enzyme that has a potent activity, which lets it to hydrolyze tissue HA, which is the key element of connective tissue. Given that, commercial hyaluronidase, when injected in areas wherein HA-based filler was placed, destroys HA and gives the possibility to adjust overcorrection and asymmetries. Although hyaluronidase has been used worldwide, only a few allergic reactions have been reported. Most of the described patients showed allergic reactions after peribulbar anesthesia for eye surgery despite the large use of HA fillers in aesthetic medicine. A 29-year-old Brazilian female patient was subjected to a 0.01 mL hyaluronidase injection (Pineda Laboratories, Sao Paulo, Brazil) in order to treat a malar hypercorrection as result of filling with HA. After about 10 minutes, she evolved with discrete erythema and edema at the injection site. A vial of 1 mL intramuscular injection of 5 mg/mL betamethasone dipropionate+2 mg/mL betamethasone disodium phosphate 2 mg/mL (BetaTrinta, Eurofarma, Sao Paulo/SP, Brazil) was immediately administered. After 1 hour, however, the patient presented an intense edema in her left hemiface, which suggested angioedema onset; this adverse event was immediately treated by injecting 4 mL of 500 mg hydrocortisone sodium succinate which helped her to clinically overcome such condition. Though, the patient was discharged to home with 40 mg/day/3 days of micronized prednisolone. A complete clinical improvement was observed in five days. In summary, side effects against hyaluronidase injections are rare in accordance with already published scientific literature; however, it is extremely important for professionals of cosmetic medicine to be an emergency conduct at their office.

Tomomitsu Miyasaka completed his PhD at Tohoku University, Japan. He is an Assistant Professor at Tohoku Medical and Pharmaceutical University, Japan. His research area is Allergy, and he is interested in determining the mechanism(s) that are responsible for the altered asthma severity by sex or related to psychological stress.

Statement of the Problem: Bronchial asthma is more severe in females than in males after puberty because of stronger Th2-oriented immune response in females. However, the mechanism of the sex difference in asthmatic immune response remains unclear. CD8+ T cells play an important role in regulating the asthma immune response through their suppressive effect on Th2 polarization within the localized lymph nodes. Theoretical Orientation: In the present study, we investigated the sex-specific effect of CD8+ T cells on the female-predominant asthmatic immune responses using a mouse model. Results: The number of eosinophil in bronchoalveolar lavage (BAL) fluid, lung Th2 cytokine levels, and IL-4 production by bronchial lymph node cells were significantly higher in wild-type female compared with male mice, whereas no such sex differences were observed between cd8α-disrupted (CD8KO) male and female mice. The adaptive transfer of wild-type male, but not female, CD8+ T cells reduced the number of inflammatory cells in the recovered BAL fluid of CD8KO male, but not female, recipient mice. Male CD8+ T cells produced higher levels of IFN-γ than female CD8+ T cells. Treatment with anti-IFN-γ antibody completely abrogated the sex difference in the suppressive activity of CD8+ T cells on IL-4 production from CD4+ T cells. However, IFN-γ receptor expression on CD4+ T cells was higher in male mice than in female mice. rIFN-γ treatment increased the proportion of IFN-γ receptor α+ CD4+ T cells in male naïve CD4+ T cells more than in female naïve CD4+ T cells. Conclusion & Significance: These results suggest that female-dominant asthmatic immune responses are induced by the reduced production of IFN-γ by CD8+ T cells and the lower expression of IFN-γ receptor on CD4+ T cells caused by exposure to IFN-γ in females compared with males.

Areum Kim is a PhD student at Jeju National University, Republic of Korea. He has research interest in Immunology and Radiobiology. He completed his BA in Nuclear and Energy Engineering at Jeju national University, South Korea from 2009-2013 and; MS in Interdisciplinary Graduate Program in Advanced Convergence Technology & Science at Jeju National University, Korea from 2013-2016.

Sargassum horneri, a species of brown macro algae, has been reported to have several health promoting effects such as anti-viral, anti-coagulant effect, and anabolic activity on bone metabolism or higher plumbum absorption. In the present study, we investigated the immunomodulatory activity of S. horneri using murine splenocytes and bone marrow cultured mast cells. S. horneri was enzymatically hydrolyzed using the celluclast (SHC). Polyphenol-rich fractions from S. horneri (SHP) were also used for assessing the anti-allergic activity in BMCMC. SHC induced the proliferation of splenocytes without cytotoxicity. Treatment with SHC led to a significant increase in the population of CD8+ T cells, CD8+CD25+ regulatory T cells, and granulocytes. SHC also increased the secretion of IFN-γ. Meanwhile, SHP didn’t show any cytotoxicity in BMCMC but significantly decreased the release of β-hexosaminidase in stimulated BMCMC. SHP also decreased the mRNA expression of IL-4, IL-6, IL-13, and TSLP in BMCMC. These findings indicate that S. horneri might modulate the Th2-type immune responses in immune-mediated disease. Therefore, S. horneri could be an excellent candidate for an anti-allergic agent.

Jinhee Cho is a PhD student at Jeju National University, Republic of Korea. He has research interest in Immunology, especially Th1/Th2 type immune responses. He completed his BA in Veterinary Medicine at Jeju National University, South Korea from 2008-2014 and; MS in Veterinary Medicine at Jeju National University, South Korea from 2014-2016.

CD244 is a member of the signaling lymphocyte activation molecule (SLAM) family and can exhibit either activating or inhibitory effect on the cytotoxic activity of cells, depending on the density of its ligand CD48 and the availability of its adaptor protein SLAM-associated protein SAP. Although the role of CD244 has mainly been studied in virus infection, its role in autoimmune disease has not been well defined. In the present study, we examined the regulation of CD244 expression in experimental autoimmune encephalomyelitis (EAE), an induced model of autoimmune disease, and determined that the role of CD48 expression on T cells was markedly changed during EAE progression. To determine the type of CD244-expressing cells affected during the EAE progression, flow cytometry analysis was performed. In the spleen of naïve mice, most of the CD244-expressing cells were NK1.1+ NK cells although NK cells were not the major population in the spleen compared to T cells. In addition, the number of CD244+ NK cells was significantly decreased in early and peak stages of EAE compared to the naïve control. At the recovery phase, the numbers of CD244+ CD4+ T cells and CD244+ CD8+ T cells were increased relative to naïve levels, respectively, while the number of CD244+ NK cells returned to the naïve level, consistent with other reports which considered CD244 as one of exhaustion markers in various diseases. In addition, high CD48 expression was associated with IL-17a production. The reduction of CD244 expression in NK cells that infiltrated into the CNS appeared more dramatic than that in the periphery. Our results suggest that CD244 expression correlates with NK cell function during EAE progression.

Inhwa Choi is working for Kyung-Hee University Hospital at Gangdong. Her specialties are in the areas of atopic dermatitis (AD) and allergic diseases, such as allergic rhinitis, asthma and allergic contact dermatitis. Her special interests include disorders of the immune system and she has devoted her time and knowledge to helping her patients reinforce and strengthen their resistance to these ailments through Korean medicine.

Patients with atopic dermatitis (AD) exhibit various symptoms, especially itching. Recently, herbal medicines (HMs) are being used in combination with antihistamines for the treatment of AD in Korea. While oral ntihistamines can alleviate itching, HMs appear to exert anti inflammatory effects with minimal side effects. However, there is little evidence regarding the effectiveness and safety of using HMs in combination with antihistamines for AD. To observe the effectiveness and safety of combination treatment with HMs and antihistamines, we performed a retrospective chart review of inpatients with AD who received this combination treatment for at least 7 days in a hospital. Of 163 inpatients, 40 met the inclusion criteria. All patients received HMs three times, and one or two antihistamines, a day after HM intake. A large proportion of patients received first-generation antihistamines. HMs comprised a mixture of an average of 20.69 different herbs in decoction. The mean total, objective, and subjective SCORing Atopic Dermatitis scores showed a significant decrease after combination treatment. Changes in the mean levels of aspartate transaminase, alanine transaminase, blood urea nitrogen, and creatinine were not statistically significant among treatments. There were no adverse events of pseudoaldosteronism or interstitial pneumonia. We observed that the short-term use of HMs in combination with oral antihistamines was safe and effective, with a low risk of adverse reactions. This study was limited by its retrospective design, and prospective studies with long-term follow-up periods are warranted to further elucidate the safety of this combination treatment for AD.