7 research outputs found
Study of the interaction between mimotope peptides of electronegative LDL with endothelial cells and macrophages.
Introdução: Dentre as diversas formas modificadas da lipoproteĂna de baixa densidade (LDL), a LDL eletronegativa, LDL (-), contribui para o processo aterosclerĂłtico, promovendo o recrutamento leucocitĂĄrio para a intima arterial e modulando a resposta inflamatĂłria. Mimotopos sĂŁo molĂ©culas que podem se ligar Ă porção variĂĄvel de um anticorpo, a enzimas ou a receptores. Os peptĂdeos mimotopos da LDL (-) sĂŁo molĂ©culas capazes de mimetizar a conformação de epĂtopos expostos apenas quando a LDL sofre modificaçÔes in vivo. Objetivo: Avaliar os efeitos de peptĂdeos mimotopos da LDL(-), previamente selecionados pela tĂ©cnica de phage display e denominados p1A3 e p2C7, em cĂ©lulas endoteliais e macrĂłfagos. Metodologia: Avaliou-se a atividade dos peptĂdeos mimotopos sobre a expressĂŁo de RNAm de genes relacionados tanto Ă ativação quanto Ă disfunção endotelial, assim como a atividade citotĂłxica e a captação dos peptĂdeos por cĂ©lulas endoteliais (HUVEC, HAEC e EA.Hy 926). Verificou-se tambĂ©m a atividade prĂł-inflamatĂłria dos peptĂdeos em cultura primĂĄria de macrĂłfagos murinos derivados da medula Ăłssea (BMDM), investigando-se a ativação do complexo proteico inflamassoma, assim como a internalização dos peptĂdeos por essas cĂ©lulas, utilizando citometria de fluxo e microscopia confocal. Resultados: As cĂ©lulas endoteliais e os macrĂłfagos internalizaram os peptĂdeos mimotopos p1A3 e p2C7. Nos macrĂłfagos houve inibição (50%) da internalização dos peptĂdeos p1A3 e p2C7 na presença de brefeldina A, indicando que esses peptĂdeos sĂŁo majoritariamente captados por endocitose independente de clatrina. Nas cĂ©lulas HAEC, observou-se 25% de inibição da captação de ambos os peptĂdeos se bloqueando o receptor scavenger LOX-1 com anticorpo anti-LOX-1. Quanto Ă atividade citotĂłxica, observou-se acentuada redução na atividade mitocondrial das cĂ©lulas endoteliais EA.Hy 926 e HAEC apenas com o peptĂdeo p1A3, nĂŁo sendo observada citotoxicidade para os macrĂłfagos tanto para p1A3 quanto p2C7 nas concentraçÔes estudadas (12,5 a 200 ug/mL). Nos macrĂłfagos, o peptĂdeo p2C7 induziu o aumento da expressĂŁo de RNAm de genes relacionados com a polarização para o fenĂłtipo M1 (iNOS, TNF-α, IL-1α) e da produção de •NO, assim como, a ativação do sinal 1 (transcricional) do complexo proteico inflamassoma, o que tambĂ©m se observou para a LDL (-). O peptĂdeo p1A3 modulou a expressĂŁo gĂȘnica da linhagem HAEC, diminuindo a expressĂŁo de RNAm dos genes relacionados Ă proliferação celular (VEGFr1 e 2, TGFβ, PDGFβ e eNOS). ConclusĂŁo: Os peptĂdeos mimotopos p1A3 e p2C7 interagem com as cĂ©lulas endoteliais e macrĂłfagos de maneiras distintas. O p2C7 induz uma resposta prĂł-inflamatĂłria em macrĂłfagos, mimetizando a LDL (-), sugerindo assim, que esse peptĂdeo possa atuar como um DAMP (padrĂŁo molecular associado ao perigo) importante para o processo inflamatĂłrio da placa aterosclerĂłtica.Introduction: Among the different subtypes of modified low-density lipoprotein (LDL), the electronegative LDL [LDL (-)] contributes to the progression of atherosclerosis, promoting leukocyte recruitment into the arterial intima and modulating the inflammatory response. Mimotopes are molecules capable of binding to the variable portion of an antibody, to enzymes or receptors. The mimotope peptides of LDL (-) are molecules mimicking the conformation of epitopes exposed only when LDL undergoes in vivo changes. Objective: To evaluate the effects of two mimotope peptides of LDL (-), previously selected the by phage display and called p1A3 and p2C7, on endothelial cells and macrophages. Methodology: We evaluated the activity of the mimotope peptides on gene mRNA expression related to both the activation and endothelial dysfunction as well as the cytotoxic activity and uptake of peptides by endothelial cells (HUVEC, HAEC and EA.Hy 926). It was also investigated the pro-inflammatory activity of the peptides on murine primary macrophage derived from bone marrow (BMDM), evaluating the activation of the protein complex inflammasome, as well as the internalization of peptides by these cells by flow cytometry and confocal microscopy. Results: The endothelial cells and macrophages internalized both peptides. In macrophages, brefeldin A inhibited the internalization of p1A3 and p2C7 around 50%, indicating that these peptides are mostly taken up by clathrin-independent bulk endocytosis. In HAEC it was observed a 25 % inhibition of the uptake of both peptides by blocking the LOX scavenger receptor with an anti-LOX-1 antibody. In relation to cytotoxicity, a reduction on mitochondrial activity was induced by p1A3 peptide only in endothelial cells (EA.Hy 926 and HAEC). No cytotoxic activity was observed for p2C7 peptide in macrophages or endothelial cells at the studied concentration range (12.5 to 200 µg/mL). In macrophages, the p2C7 peptide increased mRNA expression of genes related to the polarization to M1 phenotype (iNOS, TNF-α and IL-1α) and of •NO production, as well as, protein complex signal 1 activation (transcriptional) what was also observed for LDL (-). The p1A3 peptide modulated the expression of genes related with cell proliferation (VEGFr1 e 2, TGF?β, PDGFβ and eNOS) in HAEC. Conclusion: The mimotope peptides p2C7 and p1A3 interact with the endothelial cells and macrophages by different ways. The p2C7 induces a pro-inflammatory response in macrophages, mimicking LDL (-), thus suggesting that this peptide can act as a DAMP (Damage-associated molecular pattern) important in the inflammatory process of atherosclerotic plaque
Immunization of LDLr - / - mice with electronegative LDL mimetic peptides: a potential therapeutic strategy for atherosclerosis
Introdução: A aterosclerose Ă© uma doença inflamatĂłria crĂŽnica decorrente de alteraçÔes na parede das artĂ©rias de mĂ©dio e grande calibre e associadas a diversos fatores de risco, dentre os quais destaca-se as hiperlipidemias, ou seja, o aumento plasmĂĄtico das lipoproteĂnas, mas tambĂ©m outras comorbidades, como a SĂndrome MetabĂłlica. Entre as lipoproteĂnas, a lipoproteĂna de baixa densidade (LDL) Ă© de grande relevĂąncia na aterosclerose. Diferentes espĂ©cies de LDL modificada (LDLm) sĂŁo originadas atravĂ©s de lipĂłlise, glicação e proteĂłlise, alĂ©m da oxidação, variando em densidade e eletronegatividade, sendo melhor denominada LDL eletronegativa [LDL (-)]. Considerando as diferenças conformacionais entre a estrutura da ApoB-100 da LDL nativa e da LDL (-), em um estudo inicial, nosso grupo desenvolveu um anticorpo monoclonal (2C7) a partir da imunização de camundongos Balb/c com a LDL (-) humana. Em uma etapa seguinte foi mapeado o epĂtopo reconhecido pelo anticorpo monoclonal anti-LDL (-) atravĂ©s de phage display. O peptĂdeo ligante do anticorpo monoclonal anti-LDL (-) foi nomeado p2C7. Esse peptĂdeo nĂŁo representa regiĂ”es da sequĂȘncia linear da ApoB-100 humana, mas microdomĂnios conformacionais de epĂtopos da ApoB-100 da LDL (-), tornando-os candidatos para a imunomodulação da aterogĂȘnese. Portanto, investigar a imunomodulação induzida pelos peptĂdeo p2C7 mimĂ©ticos da LDL (-), por representar um epĂtopo imunodominante da LDL (-), poderĂĄ abrir novas perspectivas terapĂȘuticas futuras para a imunomodulação da aterosclerose. Objetivo: Avaliar a imunomodulação promovida pelo p2C7 in vivo, utilizando camundongos C57BL/6 LDLr -/- e amostras de plasma humano. Adicionalmente, no estĂĄgio (BEPE) realizado no Instituto Karolinska (dezembro de 2019 a março de 2021), investigou-se o imunometabolismo como mediador nas doenças cardiovasculares. Na parte II-A, estĂŁo descritos os resultados do estudo inicialmente proposto. Na parte II-B, apresenta-se os resultados que foram desenvolvidos posteriormente, com ampliação do escopo do projeto, abordando-se a inflamação vascular envolvida no aneurisma de aorta abdominal atravĂ©s de ferramentas de bioinformĂĄtica. Na parte II-C, sĂŁo apresentados os resultados do estudo do envolvimento da enzima indolamina 2,3 dioxigenase (IDO) na esteatohepatite nĂŁo-alcoĂłlica (NASH) e aterosclerose em camundongos ApoE-/- and ApoE/IDO/double-knockout. Metodologia: Foi avaliada a presença de anticorpos anti-p2C7 em amostras de plasma humano de indivĂduos com ou sem sĂndrome metabĂłlica. Realizamos a determinação de TNF circulante nas mesmas amostras e prosseguimos com regressĂ”es lineares associando os parĂąmetros inflamatĂłrios com os nĂveis de anticorpos anti-p2C7. Camundongos C57BL/6 LDLr -/- foram imunizados com p2C7 e os adjuvantes Alum ou Montanide ISA 720, analisando-se os tĂtulos de anticorpos contra p2C7 e LDL (-), a produção de citocinas (IL-10, IL-4, IL-2, IL-6, IFNγ, IL-17, TNFα) e cĂ©lulas secretoras de anticorpos. Camundongos C57BL/6 LDLr -/- foram tolerizados contra os peptĂdeos mimotopos, com injeçÔes intravenosas (veia caudal) e desafiados com a imunização contendo LDL (-) + Alum. Avaliou-se os tĂtulos de anticorpos contra p2C7 e LDL (-) e a produção de citocinas (TNF-α, IFNγ, IL-12, IL-6, IL-10 e MCP-1). Os camundongos foram mantidos em dieta hipercolesterolĂȘmica por 3 meses para formação da placa aterosclerĂłtica. ApĂłs este perĂodo, os camundongos foram eutanasiados, avaliando-se a formação de placa aterosclerĂłtica na artĂ©ria abdominal e arco aĂłrtico, assim como a produção de citocinas (TNF-α, IFNγ, IL-12, IL-6, IL-10 e MCP-1). Camundongos C57BL/6 LDLr -/- foram imunizados com OVA-p2C7 e, apĂłs dieta hipercolesterolĂȘmica de 3 meses para formação de placa aterosclerĂłtica, foram avaliados os parĂąmetros inflamatĂłrios e avaliada a captação de 18F-FDG no arco aĂłrtico atravĂ©s de PET/CT. Resultados: A imunização com o p2C7 (livre) nĂŁo foi capaz de induzir resposta humoral, nĂŁo se observando tĂtulos detectĂĄveis de anticorpos reativos Ă p2C7 ou LDL (-) em nenhum camundongo imunizado, assim como nĂŁo foram detectadas cĂ©lulas secretoras de anticorpos especĂficos para a LDL (-). O grupo imunizado com Alum ou Montanide + p2C7 teve aumento significativo na produção de TNF- quando comparado com os demais grupos. O protocolo de tolerização foi realizado com sucesso, visto que os camundongos tolerizados apresentaram tĂtulos de anticorpos inferiores aos controles para o epĂtopo utilizado. Apenas os camundongos tolerizados com o p2C7 apresentaram aumento significativo na produção de IL-6, IL-12, IL-10, TNF-α, IFNγ e MCP 1 apĂłs dieta hipercolesterolĂȘmica. A imunização ativa com OVA-p2C7 foi capaz de reduzir a produção de TNF induzida pela dieta hipercolesterolĂȘmica, assim como reduzir a captação de 18F-FDG. ConclusĂŁo: o epĂtopo p2C7 Ă© altamente expresso na LDL (-) de pacientes com maior risco cardiovascular. AlĂ©m disso, a imunização ativa com p2C7 tambĂ©m se mostra uma ferramenta promissora para prevenir e regular a inflamação causada pela LDL (-) no curso da aterosclerose.Introduction: Atherosclerosis is a chronic inflammatory disease resulting from changes in the wall of medium and large-caliber arteries and associated with several risk factors, among which hyperlipidemias stand out, ie, the increase in plasma lipoproteins, but also other comorbidities, such as Metabolic Syndrome. Among the lipoproteins, low-density lipoprotein (LDL) is of great relevance in atherosclerosis. Different isoforms of modified LDL (LDLm) are originated through lipolysis, glycation and proteolysis, in addition to oxidation, varying in density and electronegativity, being better called electronegative LDL [LDL (-)]. Considering the conformational differences between the ApoB-100 structure of native LDL and LDL (-), in an initial study, our group developed a monoclonal antibody (2C7) from the immunization of Balb/c mice with human LDL (-). In a next step, the epitope recognized by the anti-LDL monoclonal antibody (-) was mapped using phage display. The binding peptide of anti-LDL monoclonal antibodies (-) was named p2C7. This peptide does not represent linear sequence regions of human ApoB-100, but conformational microdomains of LDL (-) ApoB-100 epitopes, making them candidates for the immunomodulation of atherogenesis. Therefore, investigating the immunomodulation induced by p2C7 peptide mimetics of LDL (-) as it represents an immunodominant epitope of LDL (-) could open new future therapeutic perspectives for the immunomodulation of atherosclerosis. Objective: To evaluate the immunomodulation promoted by p2C7 in vivo, using C57BL/6 LDLr -/- mice, and human plasma samples. In addition, in the internship (BEPE), held at the Karolinska Institute (December 2019 to March 2021), immunometabolism as a mediator of Cardiovascular Diseases was studied. In part II-A, the results of the initially proposed study are described. In part II-B, the results that were developed later are presented, expanding the scope of the project, approaching the vascular inflammation involved in the abdominal aortic aneurysm through bioinformatics tools. In part II-C, the results of the study of the involvement of the enzyme indoleamine 2,3 dioxygenase (IDO) in non-alcoholic steatohepatitis (NASH) and atherosclerosis in ApoE-/- and ApoE/IDO/double mice are presented -knockout. Methodology: The presence of anti-p2C7 antibodies in human plasma samples with or without Metabolic Syndrome was evaluated. We measured circulating TNF in the same samples and proceeded with linear regressions associating inflammatory parameters with levels of anti-p2C7 antibodies. C57BL/6 LDLr -/- mice were immunized with p2C7 and the adjuvants Alum or Montanide ISA 720, analyzing the antibody titers against p2C7 and LDL (-), the production of cytokines (IL-10, IL-4, IL -2, IL-6, IFNγ, IL-17, TNFα) and antibody-secreting cells. C57BL/6 LDLr -/- mice were tolerized against mimotope peptides with intravenous injections (caudal vein) and challenged with immunization containing LDL (-) + Alum. Antibody titers against p2C7 and LDL (-) and cytokine production (TNF-α, IFNγ, IL-12, IL-6, IL-10 and MCP-1) were evaluated. The mice were kept on a hypercholesterolemic diet for 3 months for atherosclerotic plaque formation. After this period, the mice were euthanized, evaluating the formation of atherosclerotic plaque in the abdominal artery and aortic arch, as well as the production of cytokines (TNF-α, IFNγ, IL-12, IL-6, IL-10 and MCP -1). C57BL/6 LDLr -/- mice were immunized with OVA-p2C7 and, after a 3-month hypercholesterolemic diet for atherosclerotic plaque formation, inflammatory parameters were evaluated and 18F-FDG uptake was evaluated by PET/CT. Results: Immunization with p2C7 (free) was not able to induce a humoral response, with no detectable titers of antibodies reactive to p2C7 or LDL (-) being observed in any immunized mouse, as well as no detectable antibody-secreting cells for the LDL (-). The group immunized with Alum or Montanide + p2C7 had a significant increase in TNF-α production when compared to the other groups. The tolerance protocol was successfully performed, as the tolerized mice had lower antibody titers than controls for the epitope used. Only mice tolerated with p2C7 showed a significant increase in the production of IL-6, IL-12, IL-10, TNF-α, IFNγ and MCP 1 after a hypercholesterolemic diet. Active immunization with OVA-p2C7 was able to reduce TNF production induced by the hypercholesterolemic diet, as well as to reduce 18F-FDG uptake. Conclusion: the p2C7 epitope is highly expressed in LDL (-) of patients with higher cardiovascular risk. Furthermore, active immunization with p2C7 is also a promising tool to prevent and regulate inflammation caused by LDL (-) in the course of atherosclerosis
The role of indoleamine 2,3-dioxygenase (IDO) in the pathogenesis of non-alcoholic steatohepatitis (NASH) and atherosclerosis as comorbidities
Genetic deficiency of indoleamine 2,3-dioxygenase aggravates vascular but not liver disease in a nonalcoholic steatohepatitis and atherosclerosis comorbidity model
Nonalcoholic steatohepatitis (NASH) is a chronic liver disease that increases cardiovascular disease risk. Indoleamine 2,3-dioxygenase-1 (IDO1)-mediated tryptophan (Trp) metabolism has been proposed to play an immunomodulatory role in several diseases. The potential of IDO1 to be a link between NASH and cardiovascular disease has never been investigated. Using Apoeâ/â and Apoeâ/âIdo1â/â mice that were fed a high-fat, high-cholesterol diet (HFCD) to simultaneously induce NASH and atherosclerosis, we found that Ido1 deficiency significantly accelerated atherosclerosis after 7 weeks. Surprisingly, Apoeâ/âIdo1â/â mice did not present a more aggressive NASH phenotype, including hepatic lipid deposition, release of liver enzymes, and histopathological parameters. As expected, a lower L-kynurenine/Trp (Kyn/Trp) ratio was found in the plasma and arteries of Apoeâ/âIdo1â/â mice compared to controls. However, no difference in the hepatic Kyn/Trp ratio was found between the groups. Hepatic transcript analyses revealed that HFCD induced a temporal increase in tryptophan 2,3-dioxygenase (Tdo2) mRNA, indicating an alternative manner to maintain Trp degradation during NASH development in both Apoeâ/â and Apoeâ/âIdo1â/miceâ. Using HepG2 hepatoma cell and THP1 macrophage cultures, we found that iron, TDO2, and Trp degradation may act as important mediators of cross-communication between hepatocytes and macrophages regulating liver inflammation. In conclusion, we show that Ido1 deficiency aggravates atherosclerosis, but not liver disease, in a newly established NASH and atherosclerosis comorbidity model. Our data indicate that the overexpression of TDO2 is an important mechanism that helps in balancing the kynurenine pathway and inflammation in the liver, but not in the artery wall, which likely determined disease outcome in these two target tissues
4Th Pediatric Allergy And Asthma Meeting (Paam)
WORKSHOP 4: Challenging clinical scenarios (CS01âCS06), CS01 Bullous lesions in two children: solitary mastocytoma, S. Tolga Yavuz, Ozan Koc, Ali Gungor, Faysal Gok, CS02 Multi-System Allergy (MSA) of cystic fibrosis: our institutional experience, Jessica Hawley, Christopher OâBrien, Matthew Thomas, Malcolm Brodlie, Louise Michaelis, CS03 Cold urticaria in pediatric age: an invisible cause for severe reactions, InĂȘs Mota, Ăngela Gaspar, Susana Piedade, Graça Sampaio, JosĂ© Geraldo Dias, Miguel Paiva, MĂĄrio Morais-Almeida, CS04 Angioedema with C1 inhibitor deficiency in a girl: a challenge diagnosis, Cristina Madureira, TĂąnia Lopes, Susana Lopes, Filipa Almeida, Alexandra Sequeira, Fernanda Carvalho, JosĂ© Oliveira, CS05 A child with unusual multiple organ allergy disease: what is the primer?, Fabienne Gay-Crosier, CS06 A case of uncontrolled asthma in a 6-year-old patient, Ioana-Valentina Nenciu, Andreia Florina Nita, Alexandru Ulmeanu, Dumitru Oraseanu, Carmen Zapucioiu, ORAL ABSTRACT SESSION 1: Food allergy (OP01âOP06), OP01 Food protein-induced enterocolitis syndrome: oral food challenge outcomes for tolerance evaluation in a Pediatric Hospital, Adrianna Machinena, Olga DomĂnguez SĂĄnchez, Montserrat Alvaro Lozano, Rosa Jimenez Feijoo, Jaime Lozano Blasco, MĂČnica Piquer Gibert, MÂȘ Teresa Giner Muñoz, Marcia Dias da Costa, Ana Maria Plaza MartĂn, OP02 Characteristics of infants with food protein-induced enterocolitis syndrome and allergic proctocolitis, Ebru Arik Yilmaz, Ăzlem Cavkaytar, Betul Buyuktiryaki, Ozge Soyer, Cansin Sackesen, OP03 The clinical and immunological outcomes after consumption of baked egg by 1â5 year old egg allergic children: results of a randomised controlled trial, MerrynNetting, Adaweyah El-Merhibi, Michael Gold, PatrickQuinn, IrmeliPenttila, Maria Makrides, OP04 Oral immunotherapy for treatment of egg allergy using low allergenic, hydrolysed egg, Stavroula Giavi, Antonella Muraro, Roger Lauener, Annick Mercenier, Eugen Bersuch, Isabella M. Montagner, Maria Passioti, NicolĂČ Celegato, Selina Summermatter, Sophie Nutten, Tristan Bourdeau, Yvonne M. Vissers, Nikolaos G. Papadopoulos, OP05 Chemical modification of a peanut extract results in an increased safety profile while maintaining efficacy, Hanneke van der Kleij, Hans Warmenhoven, Ronald van Ree, Raymond Pieters, Dirk Jan Opstelten, Hans van Schijndel, Joost Smit, OP06 Administration of the yellow fever vaccine in egg allergic children, Roisin Fitzsimons, Victoria Timms, George Du Toit, ORAL ABSTRACT SESSION 2: Asthma (OP07âOP12), OP07 Previous exacerbation is the most important risk factor for future exacerbations in school-age children with asthma, S. Tolga Yavuz, Guven Kaya, Mustafa Gulec, Mehmet Saldir, Osman Sener, Faysal Gok, OP08 Comparative study of degree of severity and laboratory changes between asthmatic children using different acupuncture modalities, Nagwa Hassan, Hala Shaaban, Hazem El-Hariri, Ahmed Kamel Inas E. Mahfouz, OP09 The concentration of exhaled carbon monoxide in asthmatic children with different controlled stadium, Papp Gabor, Biro Gabor, Kovacs Csaba, OP10 Effect of vitamin D3 supplementation during pregnancy on risk of persistent wheeze in the offspring: a randomised clinical trial, Bo Chawes, Klaus BĂžnnelykke, Jakob Stokholm, Lene Heickendorff, Susanne Brix, Morten Rasmussen, Hans Bisgaard, OP11 Lung function development in childhood, Henrik Wegener Hallas, Bo Chawes, Lambang Arianto, Hans Bisgaard, OP12 Is the effect of maternal and paternal asthma different in female and male children before puberty?, Maike Pincus, Thomas Keil, Andreas Reich, Ulrich Wahn, Susanne Lau, Linus Grabenhenrich, ORAL ABSTRACT SESSION 3: Epidemiologyâgenetics (OP13âOP18), OP13 Lifestyle is associated with incidence and category of allergen sensitisation: the ALADDIN birth cohort, Sara Fagerstedt, Helena Marell Hesla, Emelie Johansson, Helen Rosenlund, Axel Mie, Annika Scheynius, Johan Alm, OP15 Maternal filaggrin mutations increase the risk of atopic dermatitis in children: an effect independent of mutation inheritance, Jorge Esparza-Gordillo, Anja Matanovic, Ingo Marenholz, Anja Bauerfeind, Klaus Rohde, Katja Nemat, Min-Ae Lee-Kirsch, Magnus Nordenskjöld, Marten C. G. Winge, Thomas Keil, Renate KrĂŒger, Susanne Lau, Kirsten Beyer, Birgit Kalb, Bodo Niggemann, Norbert HĂŒbner, Heather J. Cordell, Maria Bradley, Young-Ae Lee, OP16 Allergic multimorbidity of asthma, rhinitis and eczema in the first 2 decades of the German MAS birth cohort, Thomas Keil, Hannah Gough, Linus Grabenhenrich, Dirk Schramm, Andreas Reich, John Beschorner, Antje Schuster, Carl-Peter Bauer, Johannes Forster, Fred Zepp, Young-Ae Lee, Renate Bergmann, Karl Bergmann, Ulrich Wahn, Susanne Lau, OP17 Childhood anaphylaxis: a growing concern, Filipe Benito Garcia, InĂȘs Mota, Susana Piedade, Ăngela Gaspar, Natacha Santos, Helena PitĂ©, MĂĄrio Morais-Almeida, OP18 Indoor exposure to molds and dampness in infancy and its association to persistent atopic dermatitis in school age. Results from the Greek ISAAC II study, Athina Papadopoulou, Despina Mermiri, Elpida Xatziagorou, Ioannis Tsanakas, Stavroula Lampidi, Kostas Priftis, ORAL ABSTRACT SESSION 4: Pediatric rhinitisâimmunotherapy (OP19âOP24), OP19 Associations between residential greenness and childhood allergic rhinitis and aeroallergen sensitisation in seven birth cohorts, Elaine Fuertes, Iana Markevych, Gayan Bowatte, Olena Gruzieva, Ulrike Gehring, Allan Becker, Dietrich Berdel, Michael Brauer, Chris Carlsten, Barbara Hoffmann, Anita Kozyrskyj, Caroline Lodge, Göran Pershagen, Alet Wijga, Heinrich Joachim, OP20 Full symptom control in pediatric patients with allergic rhinitis and asthma: results of a 2-year sublingual allergen immunotherapy study, Zorica Zivkovic, Ivana Djuric-Filipovic, Jasmina JociÄ-Stevanovic, SneĆŸana Zivanovic, OP21 Nasal epithelium of different ages of atopic subjects present increased levels of oxidative stress and increased cell cytotoxicity upon rhinovirus infection, Styliani Taka, Dimitra Kokkinou, Aliki Papakonstantinou, Panagiota Stefanopoulou, Anastasia Georgountzou, Paraskevi Maggina, Sofia Stamataki, Vassiliki Papaevanggelou, Evangelos Andreakos, Nikolaos G. Papadopoulos, OP22 Cluster subcutaneous immunotherapy schedule: tolerability profile in children, Monica Piquer Gibert, Montserrat Alvaro Lozano, Jaime Lozano Blasco, Olga DomĂnguez SĂĄnchez, Rosa JimĂ©nez Feijoo, Marcia Dias da Costa, MÂȘ Teresa Giner Muñoz, Adriana Machinena Spera, Ana Maria Plaza MartĂn, OP23 Rhinitis as a risk factor for asthma severity in 11-year old children: population-based cohort study, Matea Deliu, Danielle Belgrave, Angela Simpson, Adnan Custovic, OP24 The Global Lung Function Initiative equations in airway obstruction evaluation of asthmatic children, JoĂŁo Gaspar Marques, Pedro Carreiro-Martins, Joana Belo, Sara Serranho, Isabel Peralta, Nuno Neuparth, Paula Leiria-Pinto, POSTER DISCUSSION SESSION 1: Food allergy (PD01âPD05), PD01 Allergen-specific humoral and cellular responses in children who fail egg oral immunotherapy due to allergic reactions, Marta Vazquez-Ortiz, Mariona Pascal, Ana Maria Plaza, Manel Juan, PD02 FoxP3 epigenetic features in children with cow milk allergy, Lorella Paparo, Rita Nocerino, Rosita Aitoro, Ilaria Langella, Antonio Amoroso, Alessia Amoroso, Carmen Di Scala, Roberto Berni Canani, PD04 Combined milk and egg allergy in early childhood: let them eat cake?, Santanu Maity, Giuseppina Rotiroti, Minal Gandhi, PD05 Introduction of complementary foods in relation to allergy and gut microbiota in farm and non-farm children, Karin Jonsson, Annika Ljung, Bill Hesselmar, Ingegerd Adlerbert, Hilde Brekke, Susanne Johansen, Agnes Wold, Ann-Sofie Sandberg, POSTER DISCUSSION SESSION 2: Asthma and wheeze (PD06âPD16), PD06 The association between asthma and exhaled nitric oxide is influenced by genetics and sensitisation, Björn Nordlund, Cecilia Lundholm, Villhelmina Ullemar, Marianne van Hage, Anne Ărtqvist, Catarina Almqvist, PD09 Prevalence patterns of infant wheeze across Europe, Anna Selby, Kate Grimshaw, Thomas Keil, Linus Grabenhenrich, Michael Clausen, Ruta Dubakiene, Alessandro Fiocchi, Marek Kowalski, Nikos Papadopoulos, Marta Reche, Sigurveig Sigurdardottir, Aline Sprikkleman, Paraskevi Xepapadaki, Clare Mills, Kirsten Beyer, Graham Roberts, PD10 Epidemiologic changes in recurrent wheezing infants, Herberto Jose Chong Neto, Gustavo Falbo Wandalsen, Ana Carolina Dela Bianca, Carolina Aranda, Nelson Augusto RosĂĄrio, Dirceu SolĂ©, Javier Mallol, Luis GarcĂa Marcos, PD13 A single nucleotide polymorphism in the GLCCI1 gene is associated with response to asthma treatment in children, IvanaBanic, Matija Rijavec, Davor Plavec, Peter Korosec, Mirjana Turkalj, PD14 Pollen induced asthma: Could small molecules in pollen exacerbate the protein-mediated allergic response?, Alen Bozicevic, Maria De Mieri, Matthias Hamburger, PD15 A qualitative study to understand how we can empower teenagers to better self-manage their asthma, Simone Holley, Ruth Morris, Frances Mitchell, Rebecca Knibb, Susan Latter, Christina Liossi, Graham Roberts, PD16 Polymorphism of endothelial nitric oxide synthase (eNOS) gene among Egyptian children with bronchial asthma, Mostafa M. M. Hassan, POSTER DISCUSSION SESSION 3: MechanismsâEpidemiology (PD17âPD21), PD17 Pregnancy outcomes in relation to development of allergy in a Swedish birth cohort, Malin Barman, Anna Sandin, Agnes Wold, Ann-Sofie Sandberg, PD18 Evolution of the IgE response to house dust mite molecules in childhood, Daniela Posa, Serena Perna, Carl-Peter Bauer, Ute Hoffmann, Johannes Forster, Fred Zepp, Antje Schuster, Ulrich Wahn, Thomas Keil, Susanne Lau, Kuan-Wei Chen, Yvonne Resch, Susanne Vrtala, Rudolf Valenta, Paolo Maria Matricardi, PD19 Antibody recognition of nsLTP-molecules as antigens but not as allergens in the German-MAS birth cohort, Olympia Tsilochristou, Alexander Rohrbach, Antonio Cappella, Stephanie Hofmaier, Laura Hatzler, Carl-Peter Bauer, Ute Hoffmann, Johannes Forster, Fred Zepp, Antje Schuster, RaffaeleDâAmelio, Ulrich Wahn, Thomas Keil, Susanne Lau, Paolo Maria Matricardi, PD20 Early life colonization with Lactobacilli and Staphylococcus aureus oppositely associates with the maturation and activation of FOXP3+ CD4 T-cells, Sophia Björkander, Maria A. Johansson, Gintare Lasaviciute, Eva Sverremark-Ekström, PD21 Genome-wide meta-analysis identifies 7 susceptibility loci involved in the atopic march, Ingo Marenholz, Jorge Esparza-Gordillo, Franz RĂŒschendorf, Anja Bauerfeind, David P. Strachan, Ben D. Spycher, Hansjörg Baurecht, Patricia Margaritte-Jeannin, Annika SÀÀf, Marjan Kerkhof, Markus Ege, Svetlana Baltic, Melanie C Matheson, Jin Li, Sven Michel, Wei Q. Ang, Wendy McArdle, Andreas Arnold, Georg Homuth, Florence Demenais, Emmanuelle Bouzigon, Cilla SöderhĂ€ll, Göran Pershagen, Johan C. de Jongste, Dirkje S Postma, Charlotte Braun-FahrlĂ€nder, Elisabeth Horak, Ludmila M. Ogorodova, Valery P. Puzyrev, Elena Yu Bragina, Thomas J Hudson, Charles Morin, David L Duffy, Guy B Marks, Colin F Robertson, Grant W Montgomery, Bill Musk, Philip J Thompson, Nicholas G. Martin, Alan James, Patrick Sleiman, Elina Toskala, Elke Rodriguez, Regina Fölster-Holst, Andre Franke, Wolfgang Lieb, Christian Gieger, Andrea Heinzmann, Ernst Rietschel, Thomas Keil, Sven Cichon, Markus M Nöthen, Craig E Pennell, Peter D Sly, Carsten O Schmidt, Anja Matanovic, Valentin Schneider, Matthias Heinig, Norbert HĂŒbner, Patrick G. Holt, Susanne Lau, Michael Kabesch, Stefan Weidinger, Hakon Hakonarson, Manuel AR Ferreira, Catherine Laprise, Maxim B. Freidin, Jon Genuneit, Gerard H Koppelman, Erik MelĂ©n, Marie-HĂ©lĂšne Dizier, A. John Henderson, Young Ae Lee, POSTER DISCUSSION SESSION 4: Food allergyâAnaphylaxis (PD22âPD26), PD22 Atopy patch test in food protein induced enterocolitis caused by solid food, Purificacion GonzĂĄlez-Delgado, Esther CaparrĂłs, Fernando Clemente, Begoña Cueva, Victoria M. Moreno, Jose Luis Carretero, Javier FernĂĄndez, PD23 Watermelon allergy: a novel presentation, Kate Swan, George Du Toit, PD24 A pilot study evaluating the usefulness of a guideline template for managing milk allergy in primary care, Mudiyur Gopi, Tim Smith, Edara Ramesh, Arun Sadasivam, PD26 Efficacy and safety of cowâs milk oral immunotherapy protocol, InĂȘs Mota, Filipe Benito Garcia, Susana Piedade, Angela Gaspar, Graça Sampaio, Cristina ArĂȘde, LuĂs Miguel Borrego, Graça Pires, Cristina Santa-Marta, MĂĄrio Morais-Almeida, POSTER DISCUSSION SESSION 5: Prevention and treatmentâAllergy (PD27âPD36), PD27 Allergy-protection by the lactic acid bacterium Lactococcus lactis G121: mode-of-action as revealed in a murine model of experimental allergy, Stephanie Brand, Karina Stein, Holger Heine, Marion Kauth, PD29 The relationship between quality of life and morning salivary cortisol after acute bronchiolitis in infancy, Leif Bjarte Rolfsjord, Egil Bakkeheim, Johan Alm, HĂ„vard Ove Skjerven, Kai-HĂ„kon Carlsen, Jon Olav Hunderi, Teresa LĂžvold Berents, Petter Mowinckel, Karin C. LĂždrup Carlsen, PD30 Randomised trial of the efficacy of MP29-02* compared with fluticasone propionate nasal spray in children aged â„6 years to <12 years with allergic rhinitis, Ulrich Wahn, Ullrich Munzel, William Berger, PD31 10 mg of oral bilastine in 2 to 11 years old children has similar exposure to the adult therapeutic dose (20 mg), Ulrich Wahn, RomĂĄn Valiente, Valvanera Vozmediano, John C. Lukas, MĂłnica RodrĂguez, PD33 Daily symptoms, nocturnal symptoms, activity limitations and reliever therapies during the three steps of IOEASMA programme: a comparison, Sebastiano Guarnaccia, Luigi Vitale, Ada Pluda, Emanuele DâAgata, Denise Colombo, Stefano Felici, Valeria Gretter, Susanna Facchetti, Gaia Pecorelli, Cristina Quecchia, PD34 Sensitisation to an inert aeroallergen in weaning rats and longstanding disease, in a sensitisation-tolerant and easily tolerisable rodent strain, George Guibas, Evangelia Spandou, Spyridon Megremis, Peter West, Nikolaos Papadopoulos, PD35 Bacterial and fungi exposure in school and allergic sensitisation in children, JoĂŁo Cavaleiro Rufo, Joana Madureira, InĂȘs PaciĂȘncia, LĂvia Aguiar, PatrĂcia PadrĂŁo, Mariana Pinto, LuĂs Delgado, Pedro Moreira, JoĂŁo Paulo Teixeira, Eduardo Oliveira Fernandes, AndrĂ© Moreira, PD36 Comparative study of allergy rhinitis between two populations: children vs. adults, Adriana Izquierdo Dominguez, Antonio Valero, Joaquim Mullol, Alfonso Del Cuvillo, Javier Montoro, Ignacio Jauregui, Joan Bartra, Ignacio Davila, Marta Ferrer, Joaquin Sastre, POSTER VIEWING SESSION 1: InflammationâGeneticsâImmunologyâDermatology (PP01âPP09), PP01 Immune profile in late pregnancy: immunological markers in atopic asthmaticwomen as risk factors for atopy in the progeny, Catarina Martins, Jorge Lima, Maria JosĂ© Leandro, GlĂłria Nunes, Jorge Cunha Branco, HĂ©lder Trindade, Luis Miguel Borrego, PP02 The impact of neonatal sepsis on development of allergic diseases, Secil Conkar, Mehtap Kilic, Canan Aygun, Recep Sancak, PP03 Clinical overview of selective IgE deficiency in childhood, Athina Papadopoulou, Eleni Tagalaki, Lambros Banos, Anna Vlachou, Fotini Giannoula, Despina Mermiri, PP04 Inverse relationship between serum 25(ÎÎ) vitamin D3 and total IgE in children and adolescence, Athina Papadopoulou, Stavroula Lampidi, Marina Pavlakou, Maria Kryoni, Kostas Makris, PP05, PP06, PP07 Asthma control questionnaire and specific IgE in children, Snezhina Lazova, Guergana Petrova, Dimitrinka Miteva, Penka Perenovska, PP08 Features of chronic urticaria of adolescents, Aliya Klyucharova, Olesya Skorohodkina, PP09 Cutaneous mastocytosis in children: a clinical analysis of 8 cases in Greece, Dimitra Koumaki, Alkisti Manousaki, Maria Agrapidi, Lida Iatridou, Omima Eruk, Konstantinos Myridakis, Emmanouil Manousakis, Vasiliki Koumaki, POSTER VIEWING SESSION 2: Food allergyâAnaphylaxis (PP10âPP47), PP10 Prognostic factors in egg allergy, Maria Dimou, Maria Ingemansson, Gunilla Hedlin, PP11 Evaluation of the efficacy of an amino acid-based formula in infants who are intolerant to extensively hydrolysed protein formula, Nitida Pastor, Delphine de Boissieu, Jon Vanderhoof, Nancy Moore, Kaitlin Maditz, PP12 Anaphylaxis and epinephrine auto-injector use: a survey of pediatric trainees, Adeli Mehdi, Shaza Elhassan, Carolin Beck, Ahmed Al-Hammadi, PP13 Anaphylaxis in children: acute management in the Emergency Department, Ioana Maris, Ronan OâSullivan, Jonathan Hourihane,, PP14 Understanding Cumbrian schools preparedness in managing children at risk of anaphylaxis in order to provide training and support which will create healthy and safe environments for children with allergies, George Raptis, Louise Michaelis, PP15 A new valid and reliable parent and child questionnaire to measure the impact of food protein enterocolitis syndrome on children: the FPIES Quality of Life Questionnaire (FPIESQL), Parent and Child Short Form, Audrey DunnGalvin, Matthew Greenhawt, Carina Venter, Jonathan Hourihane, PP16 An in-depth case study investigation of the experiences of teenagers and young adults in growing up and living with food allergy with emphasis on coping, management and risk, support, and social and self-identity, Evelyn OâRegan, Duncan Cronin, Jonathan Hourihane, Anna OâReilly, Audrey DunnGalvin, PP17 Cowâs milk protein allergy in Constantine. A retrospective study of 62 cases between 1996 and 2013, Foued Abdelaziz, Dounia Khelifi-Touhami, Nihad Selim, Tahar Khelifi-Touhami, PP18, PP19 Cowâs milk and egg oral immunotherapy in children older than 5 years, Pablo Merida, Ana MÂȘ Plaza, Juan Heber Castellanos, Adrianna Machinena, Montserrat Alvaro Lozano, Jaime Lozano, Olga Dominguez, Monica Piquer, Rosa Jimenez, MÂȘ Teresa Giner, PP20 Professionalsâ awareness of management of Cowâs Milk Protein Allergy (CMPA) in North Wales Hospitals, Konstantinos Kakleas, Manohar Joishy, Wendmu Maskele, Huw R. Jenkins, PP21, PP22 Anaphylaxis: the great unknown for teachers. Presentation of a protocol for schools, Mercedes Escarrer, AgustĂn Madroñero, Maria Teresa Guerra, Juan Carlos Julia, Juan Carlos Cerda, Javier Contreras, Eulalia Tauler, Maria Jesus Vidorreta, Ana Rojo, Silvia Del Valle, PP23 Challenges facing children with food allergies and their parents in out of school activity sectors, Niamh Flynn, PP24 A review of food challenges at a Regional Irish Centre, Gary Foley, Carol Harmon, John Fitzsimons, PP25 The use of epinephrine in infants with anaphylaxis, Krasimira Baynova, Ăvila Maria Del Robledo, Labella Marina, PP26, PP27, PP28 Motherâs psychological state predicts the expression of symptoms in food allergic children, Aaron Cortes, Alicia Sciaraffia, Angela Castillo, PP29 The correlation between sIgE towards tree nuts and birch pollen in a Danish Pediatric Allergy Clinic, Nanna Juel-Berg, Kirsten Skamstrup Hansen, Lars KĂŠrgaard Poulsen, PP30 Food allergy in children: evaluation of parentsâ use of online social media, Andreia Florina Nita, Ioana Valentina Nenciu, Adina Lazar, Dumitru Oraseanu, PP31 The impact of food allergy on quality of life: FAQLQ questionnaire, Rita Aguiar, Anabela Lopes, Maria J. Paes, AmĂ©lia S. Santos, M. A. Pereira-Barbosa, PP32 An unexpected cause of anaphylaxis: potato, Hatice Eke Gungor, Salih Uytun, Umit Murat Sahiner, Yasemin Altuner Torun, PP33 Is it clinical phenotype of allergic diseases determined by sensitisation to food?, Mirjana Zivanovic, Marina AtanaskoviÄ-MarkoviÄ, PP34, PP35 Prescribing adrenaline auto-injectors in children in 2014: the data from regional pediatricians, Tina Vesel, Mihaela Nahtigal, Andreja Obermayer-Temlin, Eva Ć oster KriĆŸnik, Mirjana Maslar, Ruben Bizjak, Marjeta TomĆĄiÄ-Matic, Sonja Posega-Devetak, Maja Skerbinjek-Kavalar, Mateja PredaliÄ, Tadej AvÄin, PP36 Who should have an adrenaline autoinjector? Adherence to the European and French guidelines among 121 allergists from the Allergy Vigilance Network, Guillaume Pouessel, Etienne Beaudouin, Anne M. Moneret-Vautrin, Antoine Deschildre, Allergy Vigilance Network, PP37 Anaphylaxis by Anacardium Occidentale, Marta Viñas, BartolomĂ© Borja, Nora HernĂĄndez, MÂȘ JosĂ© Castillo, Adriana Izquierdo, Marcel Ibero, PP38 Anaphylaxis with honey in a child, S. Tolga Yavuz, Ali Gungor, Betul Buyuktiryaki, Ozan Koc, Can Naci Kocabas, Faysal Gok, PP39 Evaluation of courses adopted to children on prevention, recognition and management of anaphylaxis, Tina Vesel, Mihaela Nahtigal, PP40 Symptomatic dust mites and shrimp allergy: three pediatric case reports, Filipa Almeida, Susana Lopes, Cristina Madureira, TĂąnia Lopes, Fernanda Carvalho, PP41 Poor identification rates of nuts by high risk individuals: a call for improved education and support for families, Camille Heming, Emily Garrett, Adam Blackstock, Santanu Maity, Rahul Chodhari, PP42 DAFALL: database of food allergies in the Czech Republic, Simona Belohlavkova, Eliska Kopelentova, Petr Visek, Ivana Setinova, Ivana Svarcova, PP43 Serological cross-reactivity between grass and wheat is not only caused by profilins and CCDs, Sigrid Sjölander, Nora Nilsson, Malin Berthold, Helena Ekoff, Gunilla Hedlin, Magnus Borres, Caroline Nilsson, PP44 Oil body associated proteins in children with nuts allergy. Allergens to consider in IgE-mediated nuts allergy, Loreto GonzĂĄlez DomĂnguez, Cristina Muñoz Archidona, Ana Moreira Jorge, Sergio Quevedo Teruel, Teresa Bracamonte Bermejo, Miriam Castillo FernĂĄndez, Fernando Pineda de la Losa, Luis Ăngel EcheverrĂa Zudaire, PP45, PP46 Protective effect of helicobacter pylori infection against food allergy in children, Olga Vrani, Antigone Mavroudi, Maria Fotoulaki, Maria Emporiadou, Kleomenis Spiroglou, Ioannis Xinias, PP47 Anaphylaxis pathway: A road tryp-tase to success?, Helyeh A. Sadreddini, Mia Warnes, Donna Traves, POSTER VIEWING SESSION 3: Miscell
Evaluation of a quality improvement intervention to reduce anastomotic leak following right colectomy (EAGLE): pragmatic, batched stepped-wedge, cluster-randomized trial in 64 countries
Background
Anastomotic leak affects 8 per cent of patients after right colectomy with a 10-fold increased risk of postoperative death. The EAGLE study aimed to develop and test whether an international, standardized quality improvement intervention could reduce anastomotic leaks.
Methods
The internationally intended protocol, iteratively co-developed by a multistage Delphi process, comprised an online educational module introducing risk stratification, an intraoperative checklist, and harmonized surgical techniques. Clusters (hospital teams) were randomized to one of three arms with varied sequences of intervention/data collection by a derived stepped-wedge batch design (at least 18 hospital teams per batch). Patients were blinded to the study allocation. Low- and middle-income country enrolment was encouraged. The primary outcome (assessed by intention to treat) was anastomotic leak rate, and subgroup analyses by module completion (at least 80 per cent of surgeons, high engagement; less than 50 per cent, low engagement) were preplanned.
Results
A total 355 hospital teams registered, with 332 from 64 countries (39.2 per cent low and middle income) included in the final analysis. The online modules were completed by half of the surgeons (2143 of 4411). The primary analysis included 3039 of the 3268 patients recruited (206 patients had no anastomosis and 23 were lost to follow-up), with anastomotic leaks arising before and after the intervention in 10.1 and 9.6 per cent respectively (adjusted OR 0.87, 95 per cent c.i. 0.59 to 1.30; P = 0.498). The proportion of surgeons completing the educational modules was an influence: the leak rate decreased from 12.2 per cent (61 of 500) before intervention to 5.1 per cent (24 of 473) after intervention in high-engagement centres (adjusted OR 0.36, 0.20 to 0.64; P < 0.001), but this was not observed in low-engagement hospitals (8.3 per cent (59 of 714) and 13.8 per cent (61 of 443) respectively; adjusted OR 2.09, 1.31 to 3.31).
Conclusion
Completion of globally available digital training by engaged teams can alter anastomotic leak rates. Registration number: NCT04270721 (http://www.clinicaltrials.gov)