Center for Molecular Allergology

Rome, Italy

Center for Molecular Allergology

Rome, Italy
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Pomponi D.,Center for Molecular Allergology | Di Zenzo G.,Molecular and Cell Biology Laboratory | Zennaro D.,Center for Molecular Allergology | Calabresi V.,Molecular and Cell Biology Laboratory | And 5 more authors.
British Journal of Dermatology | Year: 2013

Background Bullous pemphigoid (BP) is an autoimmune skin disease in which patient autoantibodies react with BP180 and BP230 proteins. In addition to IgG, IgE has been shown to play a role in the disease. Objectives To evaluate the feasibility of detecting IgE and IgG against the immunodominant BP180 NC16A domain (BP180) using a microarray system. Methods BP180 was immobilized on an experimental version of the ISAC® microarray (Exp96). The BP study group and the controls were all tested on the commercial ISAC 103 version and on the Exp96. IgG and IgE were measured in a single run. BP180 IgG and IgE results were compared with those using an enzyme-linked immunosorbent assay (ELISA). Results All results obtained using the IgG ELISA on the 31 patients with BP were replicated with the ISAC IgG. Five of eight BP sera tested by ELISA showed similar results with ISAC IgE. Twenty-nine (94%) and 19 (61%) of the 31 patients with BP were IgG and IgE positive to BP180, respectively, whereas four (3%) and six (4%) of 138 normal donors were IgG and IgE positive, respectively. Interestingly, the levels of IgG against BP180 detected using the ISAC system were related to the disease severity. Patients with BP showed a peculiar profile of IgE recognition toward some groups of allergens, which was absent in a group of allergic individuals. A significant, higher prevalence of hen's egg recognition was observed in patients with BP who had specific IgE to BP180. Conclusions The present preliminary study indicates that the ISAC microarray system is suitable for detecting IgG and IgE autoantibodies in patients with BP. Notably, this system allows the assessment of IgE and IgG autoantibodies at the same time, could be employed for the detection of autoantibodies to other autoantigens, and allows profiling for specific IgE to allergens. What's already known about this topic? Bullous pemphigoid (BP) is a severe autoimmune blistering disease. IgG and IgE autoantibodies to skin BP180 and BP230 autoantigens are detected by enzyme-linked immunosorbent assay (ELISA) in patients with BP. IgE seem to play a role in a subset of patients with more severe disease. What does this study add? IgG and IgE autoantibodies in BP sera were detected using the immunodominant portion of BP180 immobilized on a microarray along with allergenic molecules, with almost overlapping results compared with ELISAs. Specific IgE allergen profiles differ in patients with BP and control groups. Specific BP180 IgG and IgE are rarely detected in an allergic population. © 2012 The Authors. BJD © 2012 British Association of Dermatologists.

Araujo L.M.L.,Federal University of Paraná | Rosario N.A.,Federal University of Paraná | Mari A.,Center for Molecular Allergology
Allergologia et Immunopathologia | Year: 2016

Background: Prevalence of respiratory allergic diseases has increased worldwide. Identification of the aeroallergens involved in allergenic sensitisation is important for diagnosis, treatment and prevention. Objective: To verify the molecular pattern of sensitisation to aeroallergens in patients with allergic respiratory diseases using microarray technique for specific IgE antibody detection. Methods: Cross-sectional study of 101 children with allergic rhinitis was followed in an outpatient clinic. All patients had positive skin prick tests (SPT) to at least one of the following antigens: Dermatophagoides pteronyssinus, Blomia tropicalis, Blattella germanica, Lolium multiflorum, and dog and cat epithelium. Serum specific IgE antibodies (sIgE) to mites, animal epithelia, fungi, cockroach and pollens components were determined by ImmunoCAP ISAC. Results: sIgE to group 1 and 2 mite allergens showed higher positive rates: Der p 1 (74.2%), Der p 2 (73.3%), Der f 1 (74.2%), Der f 2 (72.3%). sIgE to animal epithelia were less frequent, Can f 1, Can f 2, Can f 3 in 4.9%, 2.9%, 1.9% respectively and Fel d 1, Fel d 2, Fel d 4 in 16.8%, 0.9% and 1.9%. respectively. Sensitisation to fungi and cockroach were rare, except for Bla g 7, to which 16.8% were positive. There was no significant recognition for tree pollens group. For grass, sIgE were detected to Cyn d 1 in 16.8%, Phl p 1 and Phl p 4 in 14.8% and 12.9%, respectively. Conclusion: Knowing that the pattern of allergic sensitisation varies according to environment and population, our results reinforce the need for local studies, using molecular-based diagnosis. © 2014 SEICAP.

Zennaro D.,Center for Molecular Allergology | Scala E.,Center for Molecular Allergology | Pomponi D.,Center for Molecular Allergology | Caprini E.,Laboratory of Molecular Oncology | And 4 more authors.
Clinical and Experimental Immunology | Year: 2012

Immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) is a rare syndrome due to a mutation in the forkhead box protein 3 gene (FOXP3) leading to an impaired regulatory T cell (T reg) activity associated both with skewed T helper type 2 (Th2) response and autoreactive phenomena. The purpose of this study was to describe a combined proteomics and genomics approach to comprehensively evaluate clinical and immunological phenotypes of patients affected by IPEX. T cell receptor (TCR)-Vβ repertoire and peripheral blood lymphocytes phenotype from three brothers affected by IPEX were studied by flow cytometry. Specific immunoglobulin (Ig)E were evaluated by means of an allergenic molecules microarray [immuno solid-phase allergen chip (ISAC)]. Total RNA was extracted and hybridized to Affymetrix oligonucleotide arrays to obtain quantitative gene-expression levels. No FOXP3 protein was detectable within CD127 -CD25 highCD4 + T cells from peripheral blood. A T cell-naive phenotype (CD62L +CD45R0 -) associated with a reduction of both CD26 and CD7 expression and a TCR-Vβ 8 and 22 family expansions were found. B lymphocytes were mainly CD5 + (B1) cells expressing a naive phenotype (tcl1 +CD27 -). The three IPEX patients had severe food allergy and specific IgE reactivity to cow's milk allergens, a hen's egg allergen and a wheat allergen. Gene expression profile analysis revealed a dysregulation associated mainly with Th1/Th2 pathways. The multiplexing evaluation reported in this study represents a comprehensive approach in the assessment of genetic conditions affecting the immune system such as the IPEX syndrome, paving the way for the development of diagnostic tools to improve the standard clinical and immunological profiling of the disease. © 2011 The Authors. Clinical and Experimental Immunology © 2011 British Society for Immunology.

Egger M.,Christian Doppler Laboratory | Alessandri C.,Center for Molecular Allergology | Wallner M.,Christian Doppler Laboratory | Briza P.,University of Salzburg | And 4 more authors.
PLoS ONE | Year: 2011

Background: Hen's egg allergy ranks among the most frequent primary food allergies in children. We aimed to investigate sensitization profiles of egg allergic patients and compare in vitro IgE reactivities of eggs from ancient chicken breeds (Araucana and Maran) with those from conventional laying hen hybrids. Methodology: Egg allergic children (n = 25) were subjected to skin prick test, double blind placebo controlled food challenge, and sensitization profiles to Gal d 1-5 were determined by allergen microarray. IgE binding and biological activity of eggs from different chicken breeds were investigated by immunoblot, ELISA, and mediator release assays. Principal Findings: We found that Gal d 1 and Gal d 2 are generally major egg allergens, whereas Gal d 3-5 displayed high sensitization prevalence only in patients reacting to both, egg white and yolk. It seems that the onset of egg allergy is mediated by egg white allergens expanding to yolk sensitization in later stages of disease. Of note, egg white/yolk weight ratios were reduced in eggs from Auraucana and Maran chicken. As determined in IgE immunoblots and mass analysis, eggs from ancient chicken breeds did not differ in their protein composition. Similar IgE-binding was observed for all egg white preparations, while an elevated allergenicity was detected in egg yolk from Araucana chicken. Conclusion/Significance: Our results on allergenicity and biological activity do not confirm the common assumption that aboriginal food might be less allergenic. Comprehensive diagnosis of egg allergy should distinguish between reactivity to hen's egg white and yolk fractions to avoid unnecessary dietary restrictions to improve life quality of the allergic child and its family. © 2011 Egger et al.

Pfiffner P.,University of Bern | Stadler B.M.,University of Bern | Rasi C.,Center for Molecular Allergology | Scala E.,Center for Molecular Allergology | And 2 more authors.
Allergy: European Journal of Allergy and Clinical Immunology | Year: 2012

Background and objective: Using an in silico allergen clustering method, we have recently shown that allergen extracts are highly cross-reactive. Here we used serological data from a multi-array IgE test based on recombinant or highly purified natural allergens to evaluate whether co-reactions are true cross-reactions or co-sensitizations by allergens with the same motifs. Methods: The serum database consisted of 3142 samples, each tested against 103 highly purified natural or recombinant allergens. Cross-reactivity was predicted by an iterative motif-finding algorithm through sequence motifs identified in 2708 known allergens. Results: Allergen proteins containing the same motifs cross-reacted as predicted. However, proteins with identical motifs revealed a hierarchy in the degree of cross-reaction: The more frequent an allergen was positive in the allergic population, the less frequently it was cross-reacting and vice versa. Co-sensitization was analyzed by splitting the dataset into patient groups that were most likely sensitized through geographical occurrence of allergens. Interestingly, most co-reactions are cross-reactions but not co-sensitizations. Conclusions: The observed hierarchy of cross-reactivity may play an important role for the future management of allergic diseases. © 2011 John Wiley & Sons A/S.

Mari A.,Center for Molecular Allergology | Ciardiello M.A.,National Research Council Italy | Tamburrini M.,National Research Council Italy | Rasi C.,Center for Molecular Allergology | Palazzo P.,Center for Molecular Allergology
Expert Review of Proteomics | Year: 2010

Conventional and innovative strategies can be exploited to identify and characterize new allergenic proteins. With the aim of obtaining suggestions for future improvements, this article describes our attempt to understand and describe some of the advantages and pitfalls of the methodologies and procedures often used in this field. The analysis includes the protein extract preparation, starting from the allergenic source, the separation of the proteins contained in a mixture and the detection, identification and characterization of IgE-binding molecules. Classic and emerging proteomic technologies, including mass spectrometry-based methodologies, Edman degradation procedure, microarray-based techniques and bioinformatics search strategies, have been explored. A comparative analysis of biochemistry-based proteomics and molecular biology strategies has also been given. © 2010 Expert Reviews Ltd.

Egger M.,Christian Doppler Laboratory | Hauser M.,Christian Doppler Laboratory | Mari A.,Center for Molecular Allergology | Ferreira F.,Christian Doppler Laboratory | Gadermaier G.,Christian Doppler Laboratory
Current Allergy and Asthma Reports | Year: 2010

Nonspecific lipid transfer proteins (LTPs) are important allergens in fruits, vegetables, nuts, pollen, and latex. Despite their wide distribution throughout the plant kingdom, their clinical relevance is largely confined to the Mediterranean area. As they can sensitize via the gastrointestinal tract, LPTs are considered true food allergens, and IgE reactivity to LTPs is often associated with severe systemic symptoms. Although Pru p 3 represents the predominant LTP in terms of patients' IgE recognition, the contribution of pollen LTPs in primary sensitization cannot be ruled out. Due to structural homology, LTPs from different allergen sources are generally IgE cross-reactive. However, sensitization profiles among allergic patients are extremely heterogeneous, and individual cross-reactivity patterns can be restricted to a single LTP or encompass many different LTPs. Molecule-based approaches in allergy research and diagnosis are important for better understanding of LTP allergy and could assist clinicians with providing adequate patient-tailored advice. © 2010 Springer Science+Business Media, LLC.

Twaroch T.E.,Medical University of Vienna | Focke M.,Medical University of Vienna | Civaj V.,Medical University of Vienna | Weber M.,Medical University of Vienna | And 7 more authors.
Journal of Allergy and Clinical Immunology | Year: 2011

Background: Trees of the family Oleaceae (olive and ash) are important allergen sources in Mediterranean countries, Northern and Central Europe, and North America. The major olive pollen allergen Ole e 1 represents the majority of allergenic epitopes in olive pollen and cross-reacts with Fra e 1, the major ash pollen allergen. Objective: We sought to develop a safe vaccine for the treatment of Oleaceae pollen allergy. Methods: We synthesized 5 peptides ranging from 32 to 36 amino acids, which covered the whole sequence of Ole e 1. The IgE and T-cell reactivity of the peptides was compared with that of Ole e 1 by means of dot blot experiments, as well as ELISA, and in proliferation assays. Rabbits were immunized with non-IgE-reactive, keyhole limpet hemocyanin-coupled peptides or Ole e 1. The reactivity of the IgG antibodies with Ole e 1 and their ability to inhibit IgE binding to nOle e 1 was evaluated by means of ELISA. Results: Only the C-terminal Ole e 1 peptide showed IgE binding, whereas the other peptides were nonallergenic. Immunization of rabbits with Ole e 1-derived peptides bound to the carrier molecule keyhole limpet hemocyanin induced in rabbits the production of Ole e 1-specific IgG antibodies, which cross-reacted with Fra e 1, and inhibited olive and ash pollen-sensitized patients' IgE binding to Ole e 1. Conclusion: Two non-IgE-binding peptides with low T-cell reactivity from the N-terminus of Ole e 1 were identified that might represent safe vaccine candidates for immunotherapy of Oleaceae pollen allergy. © 2011 American Academy of Allergy, Asthma & Immunology.

Mari A.,Center for Molecular Allergology | Alessandri C.,Center for Molecular Allergology | Bernardi M.L.,Center for Molecular Allergology | Ferrara R.,Center for Molecular Allergology | And 2 more authors.
Current Allergy and Asthma Reports | Year: 2010

IgE-mediated allergic diseases are among the most prevalent diseases worldwide. The use of extracts in the skin test and the additional use of IgE testing still represent the current basis for the diagnostic work-up. During the past 30 years, knowledge of the molecular structure of allergens has increased dramatically, and the characterization and production of allergenic molecules, as natural purified compounds or recombinant products, is allowing us to approach the allergy diagnostic work-up differently. Much of this is based on the adoption of microtechnology since the first release of a biochip for IgE detection. Its use has prompted the development of new concepts linked to the diagnosis of allergic diseases. This review describes the background of allergy diagnosis and the tools currently used for specific IgE detection. It gives insight into the most recent advancement in the field of biotechnology leading to allergenic molecule availability, microtechnology leading to the routine use of protein biochips for IgE detection, and how they should be combined with information technology. © 2010 Springer Science+Business Media, LLC.

Mari A.,Center for Molecular Allergology | Scala E.,Center for Molecular Allergology | Alessandri C.,Center for Molecular Allergology
Current Opinion in Allergy and Clinical Immunology | Year: 2011

Purpose of Review: To define the role of IgE-microarray testing in atopic dermatitis where the multifunctional environmental factors implicated in the pathogenesis of the disease deserve a complex and exhaustive approach for the finest definition of relevant allergenic triggers, if any. Recent Findings: Allergenic factors involved in atopic dermatitis have been reported in inducing atopic dermatitis. Environmental triggers, such as aeroallergens, food allergens, bacterial, and fungal antigen, and the presence of IgE to self-antigens have been described as well. The usefulness of the microarray testing has been preliminary described. Summary: As far as the number of potential triggers or worsening factors in atopic dermatitis is quite broad, we foresee a crucial role for the IgE microarray testing. The microarray testing is helpful in defining all additional reactivity the atopic dermatitis patient could have acquired in her/his allergic life, mostly related to inhalant allergens. Nevertheless, interpretation needs skills and thus this new technology should rather be reserved for the allergologist as it may lead to false conclusions if broadly used in general medicine. IgE microarray testing gives trustable results to define the nonallergic atopic dermatitis form because of the broad and comprehensive negative IgE testing. © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins.

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