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L Farioli

Publications and source records attributed to L Farioli.

At least 19 recordsLinked to original sources

Evidence for a lipid transfer protein as the major allergen of apricot.

BACKGROUND: Apricots are widely grown in Europe, and allergic reactions are becoming more common, especially oral allergy syndrome. Apricot belongs to the botanical subfamily of Prunoideae, which includes peach, the major allergen of which was identified as a 9-kd protein, a lipid transfer protein (LTP). OBJECTIVE: The aim of the study was to evaluate the IgE reactivity pattern to an apricot extract in subjects with allergic reactions to apricot, as demonstrated by a positive oral challenge response. METHODS: Thirty patients were investigated. All the patients displayed oral allergy syndrome (2 with systemic reactions) to apricot, with positive open food challenge responses, skin prick test responses, and serum-specific IgE antibodies to apricot. The IgE reactivity pattern to apricot extract was identified by using SDS-PAGE and immunoblotting. The major allergen, a 9-kd protein, was then purified by HPLC and characterized by periodic acid-Schiff stain, isoelectric point determination, and N-terminal amino acid sequencing. RESULTS: The sera from all patients allergic to apricot recognized the 9-kd protein, whereas none of the other allergens, with molecular weights from 15 to 80 kd, acted as a major allergen. The 9-kd allergen has an isoelectric point of 8.7 and is not glycosylated. Determination of the N-terminal 34 amino acid sequence showed that it belongs to the LTP family, with a 94% homology with the LTP from peach. IgE blotting of the apricot extract was completely inhibited by the 9-kd purified LTP from peach. CONCLUSIONS: The major allergen of apricot is an LTP, which is highly cross-reactive with the LTP from peach.

Adolescent↗

Technological processes to decrease the allergenicity of peach juice and nectar.

Among vegetable foods peach (Prunus persica) has been recognized as a significant cause of allergy. The protein, which is considered to be the major peach allergen, has been named Pru p 1. Because peaches are consumed both as fresh fruits and after processing to obtain peach juice, nectar, jam, syrupy peach, etc., research was carried out to identify a technological process for production of hypo- or nonallergenic peach-based products. SDS-PAGE and immunoblotting analysis of extracts prepared from four commercial peach nectars showed that the Pru p 1 was not removed, and neither was its allergenic activity decreased by technological treatments carried out for nectar production. Some treatments oriented toward a removal of or, at least, a decrease in the allergenic power were assumed and verified at laboratory scale. A variable considered was heat treatment at 121 degrees C for 10 and 30 min: this treatment was not able to decrease the allergenicity of the Pru p 1 protein. Furthermore, the protein band was still present even after 60-min reaction with two different acidic proteases. The two technological treatments that were found to decrease the major allergen of peach were chemical lye peeling of fruits and ultrafiltration of juice through membranes with suitable cutoff. On the basis of the results obtained from this research, a processing flow sheet was defined to obtain hypoallergenic or probably nonallergenic limpid juices and nectars. These products may represent, besides finished foods, intermediates to obtain various products after addition of further ingredients such as pectins, sugars, and fiber.

Beverages↗

The maize major allergen, which is responsible for food-induced allergic reactions, is a lipid transfer protein.

BACKGROUND: Cereals are the most important nutritional component in the human diet. Food-induced allergic reactions to these substances therefore have serious implications, and exhaustive diagnosis is required. Such diagnosis is still difficult because of the incomplete knowledge about major cereal allergens. In particular, few food-induced allergic reactions to maize have been reported, and no information on the allergenic proteins is available. OBJECTIVE: Having observed several anaphylactic reactions to maize, we planned a study to identify maize major allergens and cross-reactivity with other cereals, as well as to peach because the majority of patients also reacted to Prunoideae fruits. METHODS: Twenty-two patients with systemic symptoms after maize ingestion and positive skin prick test responses and serum-specific IgE antibodies to maize were selected. The IgE-reactivity pattern was identified by SDS-PAGE and immunoblotting. The major allergen identified was then purified by HPLC and characterized by mass spectrometry, determination of the isoelectric point value, and N-terminal amino acid sequencing. RESULTS: Sera from 19 (86%) of the 22 patients recognized a 9-kd protein, thus confirming this as the maize major allergen. This protein had an isoelectric point of greater than 9, a molecular mass of 9047.0 d, and no glycosylation. Determination of its N-terminal sequence showed that it was a lipid transfer protein (LTP). By using immunoblotting-inhibition experiments, we demonstrated that the LTP cross-reacts completely with rice and peach LTPs but not with wheat or barley LTPs. N-terminal sequence of the 16-kd allergen (recognized by 36% of patients) showed it to be the maize inhibitor of trypsin. This protein cross-reacts completely with grass, wheat, barley, and rice trypsin inhibitors. CONCLUSION: The major allergen of maize is an LTP with a molecular weight of 9 kd that is highly homologous with the peach LTP, the major allergen of the Prunoideae subfamily.

Adolescent↗

Clinical role of a lipid transfer protein that acts as a new apple-specific allergen.

BACKGROUND: Allergy to apple is commonly associated with birch pollinosis because the two share homologous allergens. However, some patients have apple allergy but no birch pollinosis, suggesting that there are allergens that do not cross-react with birch. OBJECTIVE: The aim of the study was to evaluate the IgE reactivity pattern to an apple extract in subjects with allergic reactions to apple, with and without birch hay fever. METHODS: Forty-three patients with oral allergy syndrome for apple and positive open food challenge, skin prick test, and serum specific IgE antibodies to apple were admitted to the study. Thirty-two had birch pollinosis (documented by specific IgE for birch) and 11 were not allergic to birch. The IgE reactivity pattern to apple extract was identified by SDS-PAGE and immunoblotting. The consistent allergen, a 9-kd protein, was then purified by HPLC and characterized by periodic acid-Schiff staining, isoelectric point, and N-terminal amino acid sequencing. RESULTS: The sera from 28% of patients allergic to apple with birch pollinosis, but from all patients allergic only to apple, recognized the 9-kd protein. This protein has an isoelectric point of 7.5 and is not glycosylated. Determination of its partial amino acid sequence showed that it belongs to the family of lipid transfer proteins, which act as major allergens in Prunoideae fruits. CONCLUSIONS: These results indicate that a lipid transfer protein is an important allergen in patients allergic to apple but not to birch pollen. The prevalent IgE reactivity to this allergen in subjects with no birch pollinosis and the physicochemical characteristics of this protein suggest that sensitization may occur through the oral route.

Adolescent↗

The major allergen of peach (Prunus persica) is a lipid transfer protein.

BACKGROUND: Allergy to fresh fruits and vegetables is mostly observed in subjects with pollinosis, especially from birch, because of cross-reacting allergens in vegetable foods and pollens. However, allergic reactions to fruits, specifically Rosaceae fruits, have been reported in subjects without pollinosis. OBJECTIVE: This study evaluated the pattern of IgE reactivity, identifying the allergen responsible in 2 groups of patients with oral allergy syndrome to peach with or without birch pollinosis. METHODS: The allergenic components of peach were detected by SDS-PAGE and immunoblotting. The major peach allergen was purified by HPLC with a cation-exchange column followed by gel filtration chromatography. Its IgE-binding capacity and its homology with the protein of the crude extract were demonstrated by immunoblotting inhibition techniques. To better characterize this allergen, periodic acid-Schiff stain and isoelectrofocusing were used. The amino acid sequencing was done with a gas-phase sequencer. RESULTS: SDS-PAGE and immunoblotting of the 15 patients allergic to peach, 8 without and 7 with birch pollinosis, showed that they all recognized a protein with a molecular weight of 9 kd. This was the only allergen recognized by patients not sensitized to pollen, whereas the birch pollen-sensitive patients had IgE binding to other allergenic proteins at higher molecular weights. The purified 9-kd protein retained its IgE-binding capacity, was negative to periodic acid-Schiff stain, and had an isoelectric point value of greater than 9. A search in the Swiss Prot Bank showed this was a lipid transfer protein, belonging to a group of molecules involved in the defensive system of plants. CONCLUSIONS: The major allergen of peach is a 9-kd protein belonging to the group of lipid transfer proteins. This is the only allergen recognized by patients allergic to peach but not sensitized to birch pollen.

Adolescent↗

Complete amino acid sequence determination of the major allergen of peach (Prunus persica) Pru p 1.

The major protein allergen of peach (Prunus persica), Pru p 1, has recently been identified as a lipid transfer protein (LTP). The complete primary structure of Pru p 1, obtained by direct amino acid sequence and liquid chromatography-mass spectrometry (LC-MS) analyses with the purified protein, is described here. The protein consists of 91 amino acids with a calculated molecular mass of 9178 Da. The amino acid sequence contains eight strictly conserved cysteines, as do all known LTPs, but secondary structure predictions failed to classify the peach 9 kDa protein as an 'all-alpha type', due to the high frequency of amino acids (nine prolines) disrupting alpha helices. Although the sequence similarity with maize LTP is only 63%, out of the 25 amino acids forming the inner surface of the tunnel-like hydrophobic cavity in maize ns-LTP 16 are identical and 7 similar in the peach homolog, supporting the hypothesis of a similar function.

Allergens↗

Sensitization to the major allergen of Brazil nut is correlated with the clinical expression of allergy.

BACKGROUND: Only a few studies have investigated the clinical role of food allergens, especially the relationship between sensitization to a given allergen and occurrence of adverse reactions when eating the relevant food item. OBJECTIVE: This study evaluated the clinical role of the allergens of Brazil nut by comparing the patterns of IgE binding in sera from 11 patients with anaphylaxis after eating Brazil nuts with those from 10 subjects with no symptoms to this food item. Both groups had specific IgE to Brazil nut. METHODS: Allergens in the in-house extract of Brazil nut were identified by SDS-PAGE/immunoblotting, the major allergen was purified by HPLC, and its N-terminal sequence was determined by a protein sequencer. RESULTS: SDS-PAGE/immunoblotting detected a number of allergenic components with molecular weights ranging from 4 to 58 kd. All sera from symptomatic patients recognized a 9-kd allergen corresponding (as established by amino acid sequencing) to a 2S albumin already described as a major allergen of Brazil nut, whereas the other allergens each bound IgE from less than 50% of sera. No sera from asymptomatic subjects showed IgE binding to the 9-kd allergen, but they did recognize components from 25 to 58 kd, which are minor allergens. CONCLUSIONS: These findings indicate that the allergen underlying clinical reactions to Brazil nut is a 2S albumin of 9 kd and that in vitro reactivity to this allergen identifies subjects who react in vivo to ingestion of this food.

2S Albumins, Plant↗

Identification of actinidin as the major allergen of kiwi fruit.

BACKGROUND: Allergic reactions to fruits and vegetables are among the most frequent food allergies in adults. Kiwi fruit (Actinidia chinensis) is commonly involved, causing local mucosal, systemic, or both types of symptoms by an IgE-mediated mechanism. In a previous study on 30 patients allergic to kiwi, we identified a major allergen of 30 kd against which all sera tested clearly reacted. Other allergens were detected at 12, 24, and 28 kd. OBJECTIVE: The aim of this study was to fully characterize the major kiwi fruit allergen of 30 kd. METHODS: Allergens were separated and purified by high-performance liquid chromatography with anion-exchange columns. The purity of the single proteins was checked by sodium dodecylsulfate-polyacrylamide gel electrophoresis, and their allergenicity was checked by immunoblotting with a pool of sera from patients allergic to kiwi. The allergens were characterized by isoelectrofocusing and amino acid sequencing, and periodic acid-Schiff stain was used to detect glycoproteins. RESULTS: Proteins of 30, 28, 24, and 17 kd were purified by high-performance liquid chromatography. IgE binding indicated the 30 kd protein, which showed an isoelectric point of 3.5, as the major allergen of kiwi. Determination of its partial amino acid sequence and comparison with the Swiss Protein Bank showed that this was actinidin, the main protein component of kiwi. The 24 and 28 kd proteins had the same N-terminal sequence, which did not correspond to any known protein. The 17 kd protein had a blocked N-terminal sequence. CONCLUSIONS: These results demonstrate that the major allergen of kiwi fruit, Act c 1, is actinidin, a proteolytic enzyme belonging to the class of thiol-proteases. Two other allergens of 24 and 28 kd appear identical on amino acid sequencing.

Allergens↗

Identification of the allergenic components of kiwi fruit and evaluation of their cross-reactivity with timothy and birch pollens.

BACKGROUND: Only a few food allergens have as yet been identified, mainly because of the difficulty of obtaining a sufficient number of patients who are clinically sensitized to a given food. This is more feasible in the case of the oral allergy syndrome (OAS), a common form of food allergy, which is especially prevalent in patients with pollinosis. OBJECTIVE: We designed a study to identify the allergens of kiwi fruit (Actinidia chinensis) by analyzing the sera of patients with OAS for kiwi and to examine the cross-reactivity of these allergens with timothy and birch pollen allergens. METHODS: Twenty-seven patients with OAS for kiwi, a positive skin prick test response and serum IgE antibody to kiwi, and a positive open kiwi challenge test result and three patients who had OAS with severe systemic symptoms, which excluded a challenge test, were included in this study. The different polypeptide components of an extract of fresh kiwi were separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis and analyzed by IgE immunoblotting with sera from these patients. Cross-reactivity with the two pollen extracts was assessed by inhibition of the immunoblots with pooled and individual patients' sera. RESULTS: Twelve IgE-binding components with molecular weights ranging from 12 to 64 kd were identified in the kiwi extract, but only a 30 kd component acted as major allergen, being recognized by sera of 100% of these patients. Inhibition of kiwi immunoblots with timothy and birch pollen extracts demonstrated strong cross-reactivity with some of the kiwi allergens, suggesting complete identity between certain food and pollen allergens; whereas others, particularly the 30 kd allergen, were only partially inhibited, suggesting much weaker cross-reactivity. CONCLUSIONS: Kiwi fruit contains a large number of allergens widely cross-reacting with allergens in grass and birch pollen extracts. Nevertheless, the major allergen at 30 kd appears to be specific for kiwi.

Allergens↗

Allergenic cross-reactivity among peach, apricot, plum, and cherry in patients with oral allergy syndrome: an in vivo and in vitro study.

BACKGROUND: Oral allergy syndrome in response to fruits and vegetables frequently occurs as clusters of hypersensitivity to members of the same botanical family, for which the immunologic basis lies in a number of common allergens, most of them still unidentified. OBJECTIVE: This study was designed to assess the in vivo and in vitro cross-reactivity between fruits of the Prunoideae subfamily (i.e., peach, cherry, apricot, and plum) and to identify their major allergens and the cross-reactivity of the peach extract with grass and birch pollen. METHODS: The in vivo study was conducted by skin prick tests and open food challenges with fresh fruits in 23 patients with oral allergy syndrome for peach and positive skin prick test and RAST results for the other Prunoideae. In vitro sodium dodecylsulfate-polyacrylamide gel electrophoresis was followed by immunoblotting and immunoblotting-inhibition. RESULTS: A 13 kd component was identified as the only major allergen common to all the Prunoideae, the other major allergens were found at 14 kd in peach and at 30 kd in cherry. Immunoblotting inhibition showed wide cross-reactivity within the Prunoideae, whereas grass and birch pollen partially inhibited the peach blotting. CONCLUSIONS: Clinical cross-reactivity to Prunoideae is essentially due to a common 13 kd IgE-binding component, which seems to be the most important major allergen of this subfamily, not shared with grass and birch pollen.

Allergens↗

A double-blind, placebo-controlled study of immunotherapy with an alginate-conjugated extract of Parietaria judaica in patients with Parietaria hay fever.

A double-blind, placebo-controlled study was conducted to evaluate the efficacy and safety of immunotherapy (IT) with a partially purified alginate-conjugated extract of Parietaria judaica (Conjuvac Parietaria, Dome/Hollister-Stier) in patients suffering from rhinoconjunctivitis caused by Parietaria pollen. Eighteen patients (10 women, 8 men, mean age 35 years) received active treatment and 17 (10 women, 7 men, mean age 42.5 years) received placebo. Actively treated patients had significantly lower nasal symptom/medication scores (running nose P = 0.0087 and sneezing P = 0.048) during the Parietaria pollen season. Significant decreases in specific skin (P < 0.01), nasal (P < 0.05), and conjunctival (P < 0.01) reactivity to the Parietaria extract and significant increases of specific IgG (P < 0.001), IgG1 (P < 0.001), and IgG4 (P < 0.001) in actively treated patients, but not in placebo, were found. IT was well tolerated, the active extract inducing five mild systemic reactions (four rhinitis and one urticaria) and placebo two (rhinitis). A significant correlation was found between low skin reactivity and high specific IgG (P = 0.0002) and IgG4 (P = 0.036). These findings indicate that IT with a partially purified P. judaica extract is an effective and safe treatment for Parietaria pollen allergy. The correlation between low immediate skin reactivity and high specific IgG and IgG4 suggests that, at least in the studied cutaneous model, these antibodies may exert a blocking effect.

Adjuvants, Immunologic↗

IgE-mediated allergy from vegetable allergens.

Allergy to fresh fruits and vegetables is IgE-mediated. Its main clinical features are local, such as oral pruritus and swelling of lips and tongue but systemic symptoms such as urticaria, asthma, or anaphylactic shock may occur. Clinical associations with allergic rhinitis due to cross-reactive antigens of pollens and foods are frequent.

Adolescent↗

Clinical evaluation of CAP System and RAST in the measurement of specific IgE.

We investigated the diagnostic value of a new in vitro test, Pharmacia CAP System (Pharmacia Diagnostics AB, Uppsala, Sweden), for the quantitative measurement of allergen-specific IgE antibodies by comparison with RAST in 2 groups of patients, 71 atopic and 48 non-atopic. In the last 20 years RAST has supplied a good diagnostic tool, but this test presents some problems, the main one being sensitivity. The new test has a solid phase able to bind even very small amounts of specific IgE and an anti-IgE tracer with very low cross-reactivity with other immunoglobulins, thus presenting more favourable conditions. From the analysis of our results, Pharmacia CAP System gave higher sensitivity (94% compared to 88% of RAST) with no loss of specificity (96% for both tests). The reliability of these results is ensured by the proper selection of patients who were all suffering from pollinosis and were clinically diagnosed as certainly hypersensitive to a single pollen. A positive trend was found between severity of asthma and levels of specific IgE for timothy. Pharmacia CAP System appears to identify a larger number of atopic patients than RAST.

Adolescent↗

A double-blind study of hyposensitization with an alginate-conjugated extract of Dermatophagoides pteronyssinus (Conjuvac) in patients with perennial rhinitis. II. Immunological aspects.

In a 2-year double-blind placebo controlled study an immunological evaluation was carried out on 33 patients (15 males, 18 females, mean age 29.2 years) with mite-induced perennial rhinitis who were submitted to specific immunotherapy (IT) with an alginate-conjugated extract of D. pteronyssinus. The behaviour of IgE, IgG, IgG1 and IgG4 antibodies specific to D. pteronyssinus and its major allergen Der p1 was characterized by assessment of their changes in serum, and changes in IgG in nasal secretions during the treatment. The placebo-treated patients did not show any significant variation in the levels of specific antibodies, while in the actively treated patients we found: a statistically significant decrease (P less than 0.005) of specific IgE, a statistically significant increase of specific IgG (P less than 0.005), IgG1 (P less than 0.005) and IgG4 (P less than 0.005) in serum and a statistically significant increase (P less than 0.001) of specific IgG in nasal secretions. The IgG response showed an early relative predominance of the IgG1 subclass and a late absolute predominance of IgG4 subclass, that confirmed the model of IgG4 restriction in prolonged allergen stimulation. No correlation was found between immunological and clinical data.

Adjuvants, Immunologic↗

[Allergy to Hymenoptera venom in children. Diagnosis and therapy].

We studied 52 children, 3 to 15 years of age, suffering from allergic reactions following Hymenoptera stings. The diagnosis of allergy was based on history, results of skin tests and RAST performed by purified venoms of Apis mellifera, Vespula species and Polistes species. The history of children with multiple stings of the same insect showed an essential favourable prognosis, thus suggesting to submit to specific immunotherapy only children with severe reactions. For these patients it is necessary to perform therapy only in specialized Centres, because of the high frequency of adverse reactions which we observed in 33% of our treated patients.

Adolescent↗

[Epidemiological and clinical study on bee venom allergy among beekeepers].

A randomized population of 222 beekeepers from Lombardy (203 males, 19 females, of mean age 42.5 years) was studied to determine the frequency of allergic reactions to bee sting. The type of reactions, the clinical evolution at the subsequent stings and the risk factors concerning the development of allergy (presence of venom specific IgE, number of stings in a year, atopy, age) were evaluated. It was found that 170 beekeepers never presented reactions to stings while 52 (23.4%) showed allergic reactions consisting in 31 large local reactions and 21 systemic reactions; of these, 3 (5.7%) were life-threatening. In the group of beekeepers with allergic reactions at the subsequent stings, 26 (50%) showed a spontaneous loss of reactivity, 16 (30.8%) presented persistent, but unchanged in severity, reactions and 10 (19.2%) had a worsening of symptoms. Specific honey bee venom IgE levels (measured by means of RAST) were significantly lower in immune beekeepers when compared with the group with allergic reactions (p less than 0.01) and in beekeepers with previous allergy when compared to the ones with persistent reactions (p less than 0.05). We also found significant differences about the number of stings received in a year by beekeepers with persistent allergic reactions (17.5 stings), beekeepers with previous allergy (89.8 stings) and immune subjects (126.9 stings). On the contrary, no significant differences were detected about the age and the presence of atopy. These results suggest that practice of bee-keeping induces a relatively high incidence of allergic reactions but with a trend to the spontaneous improvement of symptoms and a low incidence of severe reactions.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗