Biomedical subjects
D Roncarolo
Publications and source records attributed to D Roncarolo.
A case of sesame seed-induced anaphylaxis.
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Intraoral and respiratory allergy to Eruca sativa (Cruciferae family) in a subject with sensitization to pollen allergens.
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Dot immunobinding assay for detection of mite antigens in house-dust samples.
A new test was developed specifically to detect mite antigens in house-dust. It uses a nitrocellulose dipstick spotted with specific antimite antibodies that act as a capture matrix; the same antibodies act as a detecting reagent when conjugated with colloidal dye particles. Aclotest is a 1-step assay, where a spotted dipstick is placed in a tube containing the detecting reagent and the house-dust sample. No instrumentation or previous extraction procedure of the sample is required, and the test response is visible as a colored spot, after 1 h incubation. The sensitivity and specificity of the new test were compared with those of Acarex and Der p1/Der f1 ELISA tests.
Cross-reactivity between latex and sweet pepper due to prohevein.
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Exercise-induced egg anaphylaxis.
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Monomeric chemically modified allergens: immunologic and physicochemical characterization.
Allergenic extracts (Der p, grass, and Parietaria) or single allergens such as Par j I (the major allergen of Parietaria) and ovalbumin (OA), a food allergen widely used in animal models, were chemically modified by reaction with potassium cyanate (KCNO), which transforms the epsilon-amino group of the lysine of proteinaceous allergens into ureido groups. KCNO-modified (carbamylated) allergens have low allergenic potency, as demonstrated in vitro (RAST inhibition) and in vivo (passive cutaneous anaphylaxis). When used to immunize rabbits, carbamylated allergens still induce IgG antibodies able to cross-react with native allergens (immunoblotting experiments). An interesting feature distinguishing carbamylated allergens from other chemically modified allergens is the preservation of the native monomeric dimension as demonstrated by SDS-PAGE analysis. Results are discussed from the perspective of clinical application of carbamylated allergens.
Modified par j I allergen from P judaica pollen and its rate of absorption in rats.
Polymerized allergens (allergoids) have been introduced in the immunotherapy of allergic disease in order to reduce the risk of side effects. However, their high molecular weight can be a limit, particularly when they are administered by a route involving passage through the mucosal barrier. We describe a simple procedure aimed at developing an original modified allergen with significantly less allergenic potential (intended as human IgE-binding capacity) but preserving the monomeric nature of the molecule. Par j I, the major allergen of Parietaria judaica pollen, was purified by a combination of monoclonal antibodies and affinity chromatography. Par j I allergen was then modified by reaction with potassium cyanate (KCNO), and compared with the native allergen to evaluate its allergenic potency (RAST-inhibition) and molecular weight (SDS-PAGE). Modified allergen showed significantly lower allergenic potency but kept its original molecular weight, making it particularly suitable for buccal (sublingual) administration. To study the adsorption profile, modified Par j I was radiolabeled and administered intravenously and sublingually to normal rats. The prospects for clinical application of the modified allergen are discussed.
Dot immunobinding assay as a new diagnostic test for human hydatid disease.
Bovine and human hydatid antigens collected from hepatic cysts and characterized by SDS-PAGE immunoblotting show similar patterns. The bovine hydatid antigen has been used to develop a simple and fast in vitro diagnostic assay for human hydatidosis. This method, named HA-DIA (hydatid antigen dot immunobinding assay), consists of incubation of a serum sample with a textile colloidal dye (pink) and a nitrocellulose stick to which the hydatid antigen has been bound. The presence of parasite-specific antibodies leads to dyeing of the stick reactive area, and a coloured spot appears. HA-DIA sensitivity and specificity have been studied in comparison with RAST-IgE and ELISA-IgG by testing 17 sera of patients with hydatid disease and 36 control sera from patients affected with other parasitic and non-parasitic diseases. HA-DIA showed positive results in all the patients' sera and in none of the control sera. Correlation with ELISA--IgG and RAST-IgE was significant. HA-DIA has been demonstrated to be of good predictive value, allowing a speedy diagnosis of hydatid disease. In view of its simplicity, not requiring any laboratory instruments, it is particularly suitable for large-scale field screening.
Lipid transfer protein: a pan-allergen in plant-derived foods that is highly resistant to pepsin digestion.
BACKGROUND: Lipid transfer proteins (LTPs) are stable and highly conserved proteins of around 10 kD. They have recently been identified as allergens in fruits of the Rosaceae family. OBJECTIVE: The aim of this study was to investigate whether the highly conserved structure of LTPs justifies a designation as a true pan-allergen, and to study the role of protein stability in allergenicity. METHODS: Thirty-eight patients with a positive skin prick test to Rosaceae fruit extracts were characterized by interviews and skin prick tests. To investigate IgE cross-reactivity between Rosaceae and non-Rosaceae LTPs, RAST and RAST inhibition as well as ELISA and ELISA inhibition were performed, using whole food extracts and purified natural and recombinant LTPs. To address the role of protein stability in the allergenicity of LTP, fruit extracts and LTPs were digested with pepsin. RESULTS: IgE antibodies to Rosaceae LTPs cross-reacted with a broad range of non-Rosaceae vegetable foods. Inhibition studies with purified natural and recombinant LTPs confirmed the role of LTP in this cross-reactivity. Many of the patients with this type of cross-reactive IgE antibodies had a clinical food allergy. In contrast to the typical birch Rosaceae cross-reactive patients, the oral allergy syndrome was frequently accompanied by more severe and systemic reactions. IgE reactivity to LTP was shown to be resistant to pepsin treatment of the allergen. CONCLUSION: LTP is a true pan-allergen with a degree of cross-reactivity comparable to profilin. Due to its extreme resistance to pepsin digestion, LTP is a potentially severe food allergen.
Cross-reactivity between IgE-binding proteins from Anisakis simplex and Dermatophagoides pteronyssinus.
An association was found between Anisakis simplex (As) and Dermatophagoides pteronyssinus (Dp) sensitization. One recent study shows a cross-reactivity between As and Dp and tropomyosin (tr)is suspected as being one of the proteins responsible of this cross-reaction. The aim of our study was: 1) to confirm the cross-reactivity between Dp and As; 2) to determine the importance of tr in this cross reaction. SDS-PAGE analysis of Dp and As (metabolic and somatic) extracts was carried out. Then an IgE immunoblotting test using serum from a patient who had specific IgE only to Dp and As and immunoblotting inhibition experiments using Dp extract and tr as inhibitors were performed. We found that patients serum reacted: 1) against larval As antigens with a molecular weight (mw) of 25 kilodalton (kD) and a mw > than 100 kD, 2) against various metabolic As antigens with a mw > than 100 kD, a mw ranging approximately from 35 to 50 kD, and a mw around 20 kD, and 3) against Dp proteins with mw between 35 and 55 kD. Preincubation of patient's serum with Dp extract caused the disappearance of reactivity against antigens with a mw > than 100 kD in both larval and metabolic As extracts and against proteins with mw ranging approximately from 35 to 50 kD in the metabolic As extract. Preincubation of patients serum with As extract caused the disappearance of reactivity against antigens with mw between 35 and 55 kD in the Dp extract. Pre-incubation of patients serum with tr did not induce any change in the immunoblotting profile. The results show that 1) cross-reactive components between Dp and As are some proteins with a mw ranging approximately from 35 to 50 kD and with a mw > than 100 kD, and 2) tr is not involved in cross-reactivity between As and Dp.
Anaphylaxis to pine nuts and immunological cross-reactivity with pine pollen proteins.
Despite the wide use of pine nuts, the fruit of Pinus pinea, only a few reports of allergic reactions to them have been published. We present herein a case of food allergy to pine nuts in a patient who showed no clinical symptoms to pine pollen despite the presence in her serum of specific IgE antibodies. In order to verify whether the reaction against pine nuts was IgE mediated, specific IgE against pine nuts and pollen were evaluated by skin-prick test, prick by prick and RAST. Immunoblotting and immunoblotting-inhibition were used to evaluate the allergenic components of both extracts and their cross-reactivity. Prick by prick with fresh pine nuts and RAST with pine nut and pine pollen extracts showed that the patient had high levels of specific IgE against both extracts. Immunoblotting experiments showed the presence in serum of IgE antibodies against several components in pine nuts and pollen. Immunoblotting-inhibition experiments demonstrated the presence of some cross-reacting components. These data confirm the existence of food allergy induced by pine nuts. This sensitization to pine nuts developed with no symptoms of pine pollinosis. Development of pollinosis may require a longer time of exposure to allergens. Based on the cross-reactivity between pine nut and pine pollen extracts, cosensitization to these two allergens could be possible.