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Passalacqua G.,University of Genoa | Melioli G.,Laboratorio Centrale Of Analisi | Bonifazi F.,Allergy Unit | Bonini S.,The Second University of Naples | And 7 more authors.
Allergy: European Journal of Allergy and Clinical Immunology

Background The IgE response is directed against specific components from an allergenic source. The traditional diagnostic methods use whole extracts, containing allergenic, nonallergenic and cross-reactive molecules. This may pose diagnostic challenges in polysensitized patients. Microarray techniques detect specific IgE against multiple molecules, but their value in term of additional information and economic saving has not been yet defined. Objective We assessed the additional diagnostic information provided by an allergen microarray in a large population of polysensitized subjects. Methods In this multicentre study, allergists were required to carefully record diagnosis and treatment of consecutive patients referred for asthma/rhinitis, using the standard methodology (history, skin prick test, IgE assay). Then, a microarray allergen assay was carried out. Clinicians were required to review their diagnosis/treatment according to microarray results. Results 318 allergic patients (30% reporting also nonrespiratory symptoms) and 91 controls were enrolled. The clinicians reported at least one additional information from the microarray in about 60% of patients, this resulting in therapeutic adjustments. In 66% of patients IgE to pan-allergens were detectable, being this clinically relevant in 38% of patients with polysensitization to pollens. Conclusion Microarray IgE assay represents an advancement in allergy diagnosis, as a third-level approach in polysensitized subjects, when the traditional diagnosis may be problematic. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd. Source

Rossi R.E.,Allergy Unit | Incorvaia C.,Allergy Pulmonary Rehabilitation | Moingeon P.,Stallergenes SA | Frati F.,Stallergenes | And 3 more authors.
Clinical and Molecular Allergy

Background: Some studies reported the possible induction of food allergy, caused by neo-sensitization to cross-reacting allergens, during immunotherapy with aeroallergens, while other studies ruled out such possibility.Objectives: The aim of this study was to evaluate the development of neo-sensitization to Pen a 1 (tropomyosin) as well as the appearance of reactions after ingestion of foods containing tropomyosin as a consequence of sublingual mite immunization.Materials and methods: Specific IgE to Tropomyosin (rPen a 1) before and after mite sublingual immunotherapy in 134 subjects were measured. IgE-specific antibodies for mite extract and recombinant allergen Pen a 1 were evaluated using the immunoenzymatic CAP system (Phadia Diagnostics, Milan, Italy).Results: All patients had rPen a 1 IgE negative results before and after mite SLIT and did not show positive shrimp extract skin reactivity and serological rPen a 1 IgE conversion after treatment. More important, no patient showed systemic reactions to crustacean ingestion.Conclusions: Patients did not show neo-sensitization to tropomyosin, a component of the extract (namely mite group 10) administered. An assessment of a patient's possible pre-existing sensitisation to tropomyosin by skin test and/or specific IgE prior to start mite extract immunotherapy is recommended. © 2010 Rossi et al; licensee BioMed Central Ltd. Source

Rossi R.E.,Rete di Allergologia Regione Piemonte | Melioli G.,Laboratorio Centrale Of Analisi | Monasterolo G.,Laboratorio Analisi Ospedale Of Savigliano E Fossano | Harwanegg C.,Genosense | And 3 more authors.
European Annals of Allergy and Clinical Immunology

Background: The micro-array techniques for the detection of specific IgE has improved the diagnostic procedures for allergic diseases. This method also allows to define sensitization profiles from an epidemiological point of view.We studied the sensitisation pattern in a population of polysensitized patients with respiratory allergy, living in a restricted geographical area in the north-west Italy. Methods: Consecutive patients with asthma/rhinitis, living in the province of Cuneo, and having at least two positive skin prick test for non related aeroallergens were studied by a microarray (Phadia, Milan Italy) which allowed to detect specific IgE against 103 different allergen components. Results: The 70 patients included had specific IgE towards a mean of 4.3 allergens/ patient (range 2-12 allergens). Concerning pollens, 63 (90%) had specific IgE to at least one genuine grass pollen allergen, 32 (45.7%) had Ole e 1 specific IgE antibodies, although olive tree is not present in the area. A relevant percentage of sensitization to mite was found (47,1%). True co-sensitisation to grass-pollen allergens/Bet v 1/Ole e 1 was observed in 15 individuals (21.4%). Pru p 1, resulted to be a sensitizing allergen in 23 patients (32.85%), 4 of whom were co-sensitised to Pru p 3 and/or Art v 3. Conclusion: A detailed knowledge of the sensitisation pattern may have relevant implications for the prescription of specific immunotherapy. Moreover, sensitisation to PR-10 (or profilin), frequently associated to oral allergy syndrome, in some cases could hide the sensitisation to LTPs which are clinically more relevant. Source

Montalto E.,Messina University | Mangraviti S.,Laboratorio Centrale Of Analisi | Costa G.,Messina University | Carrega P.,Laboratorio Of Immunologia Clinica E Sperimentale | And 8 more authors.
Immunology Letters

Seroma is a frequent complication of breast cancer surgery, the etiology of which remains indefinite. It represents a subcutaneous accumulation of fluid frequently reported after surgical procedures such as axillary lymph node dissection.Despite previous studies have associated seroma fluid to an inflammatory exudate, the surgical removal of draining lymph nodes may indicate that seroma might not represent a mere exudate but rather an accrual of lymph drained from tributary tissues. To verify this hypothesis, seromas were collected at different intervals of time in patients operated upon for axillary lymph node removal. Fluids were analyzed in details by flow cytometry and biochemical assays for their cellular content and for their molecular features and relevant cytokine content.Lymphocytes and other peculiar blood mononuclear cells were present, while erythrocytes, platelets and granulocytes were absent or extremely rare. The protein concentration resulted lower (median 64%) than in peripheral blood. However, specific proteins related to locoregional tissues resulted highly concentrated (e.g. up to 500% for ferritin and 300% for lactate deydrogenase and exclusive presence of interleukin-6) whereas all enzymes and proteins synthesized in the liver or other organs (e.g. alkaline phosphatase, ALT, γGT, prealbumin, transferrin, ceruloplasmin, C3 and C4, α2 macroglobulin from liver; apolipoproteins from liver and gut; amylase and lipase from pancreas) were represented in reduced concentrations, thus ruling out that seroma proteins derive directly from blood serum. As a whole, this comprehensive cytological and molecular analysis provided evidences that seroma is constituted by serum ultrafiltrated-derived extracellular fluid of regions located upstream of removed lymph nodes. This fluid is then enriched by proteins and cells collected in the drained regions. Remarkably, seroma fluids collected in the same patient at different time points (up to 50 days following surgery) displayed similar biochemical features, clearly indicating that fluid composition was not significantly affected by post-surgical locoregional flogosis. Finally, the period of seroma formation indicates that lymph accumulates in the axillary region during the interval of time needed for afferent lymphatic vessels to re-anastomose with the efferent ducts.Therefore, seroma fluid represents a font of biological material suitable for investigating the biology of breast cancer, healing tissues and lymph. © 2010 Elsevier B.V. Source

Mistrello G.,Lofarma S.p.A | Agazzi A.,Laboratorio Centrale Of Analisi | Melioli G.,Laboratorio Centrale Of Analisi
European Annals of Allergy and Clinical Immunology

Limonium tataricum (Lt) is a plant belonging to the family of Plumbaginaceae. The role of this family and in particular, that of dried flowers (but not of the pollen) in occupational allergy has already been described.We have observed a farmer with asthma occurring in the presence of fresh flowers. Standard methacoline test demonstrated that the patient was a true asthmatic. The allergenicity of Lt pollen was thus investigated. Skin prick tests (SPT) were carried out using both standard allergens and the Lt extract and the patient's mucosal reactivity was evaluated by nasal provocation test with the pollen extract. In vitro studies were also performed on the patient's serum by evaluating routine specific anti-allergen IgE on raw extracts and on Microarray Allergen Chip (ISAC). Finally, the raw extract of the fresh Lt pollen was also used in ELISA inhibition test, immunoblotting and Basophil Activation Test (BAT). The specific sensitization was demonstrated by Skin Prick test and nasal provocation test. The sensitization was also confirmed by specific IgE and by in vitro activation of basophils in the presence of the pollen. By using RAST inhibition test, the presence of cross-reactivity with other pollens was ruled out. According to our results, Lt extracts contain an allergenic activity not only as dried flowers, but also as fresh pollen. For its role in occupational asthma, this allergen should be included in any allergy screening at least in farmers or in the flower industry employers. Source

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