Le platane

[3] - Skypala I, Calderon M, Leeds A, Durham S. Pollen-food syndrome in United Kingdom subjects - which aeroallergens are commonly involved ? Allergy 2007;62(suppl. 83):358

Pollen food syndrome (PFS) is common in individuals with birch pollen sensitivity who present with hay fever symptoms in the spring. The aim of this study was to evaluate which pollen(s), apart from birch, are associated with PFS in UK-based subjects, and whether aero-allergen sensitivities differ in those subjects with springtime hay fever with/without associated PFS. Adult subjects reporting hay fever symptoms between March and May were included in the study. All subjects underwent skin prick testing with tree (birch, oak, beech, plane and three trees ˜ birch, alder and hazel), timothy grass, mugwort, latex, cat and house dust mite (D. pteronyssinus) allergens (ALK Abelló Denmark). A diagnosis of PFS was established for each subject using a combination of results from a standardised medical assessment, prick by prick testing with suspect foods and oral food challenge. 119 subjects completed the study; 92% were sensitised to one or more aeroallergens. Although all subjects reported spring and/or summer time hay fever, only 85% of the cohort were sensitised to pollen. Grass was the commonest sensitising pollen allergen (68%), followed by birch (64%), oak and three trees (53%), beech (34%), plane (26%) and mugwort (18%). Approximately half of the cohort were sensitised to cat and/or house dust mite, but less than 10% to latex. Subjects diagnosed with PFS had a significantly greater number of positive tests for birch, oak, beech (p<0.001) and grass (p<0.05) than those without PFS, but there was no significant difference between the groups for the other aeroallergens. Polysensitisation to pollen allergens occurred in 93% of the group diagnosed with PFS compared to 37% of the group without PFS. No aeroallergen sensitivities were individually or collectively significant predictors of the diagnosis of PFS although a combination of birch, grass and oak gave the best standardised beta coefficient. UK subjects with springtime hay fever are most likely to be sensitised to birch, grass and oak. The frequency of sensitivities to these allergens is greater in those who additionally have PFS. In contrast, subjects with/without PFS do not differ in their rate of sensitisation to cat and house dust mite. Sensitisation to pollen cannot predict who with springtime hay fever will have PFS, but those with PFS are more likely to be co-sensitised to several tree and grass pollens.

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[4] - Bousquet PJ, Gallega MP, Dhivert-Donnadieu H, Demoly P. Latex is not essential in a standardized skin prick test battery. Allergy 2005;60:407-408

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[5] - Bousquet J, Cour P, Guerin B, Michel FB. Allergy in the Mediterranean area I. Pollen counts and pollinosis of Montpellier. Clin Allergy 1984;14:249-258

The climatic conditions of the Mediterranean area result in vegetation and pollen very different from that of the other parts of Europe. The pollen content of the atmosphere of Montpellier, southern France, was examined using a filter sampler which was shown to be more efficient than most of the current devices for air sampling. Pollen counts were subsequently compared with pollinosis of patients born and living in and around Montpellier. The mean annual pollen counts showed that grass pollens and Cupressaceae pollens (cypress and juniper) are the highest. Some Mediterranean pollens (Oleaceae, London plane, Parietaria) are also important. Plantain and oak pollens are also present in relatively large amounts. Grass pollen allergy was found to be present in 86.5% of pollen-allergic patients. It was followed by plantain, Parietaria, Oleaceae, London plane and Cupressaceae pollens which were allergenic in 13-36% of pollen-allergic patients. Oak and pine pollens were present in large quantities in the counts but few persons were sensitive to oak and none to pine. By contrast, some patients had positive skin tests to alfalfa, red clover, acacia and lime tree pollens though these pollens were almost absent from the counts. In a few cases local sources of these pollens could account for the positive skin tests but cross-sensitivities could also occur. In summary, pollinosis of the Northern Mediterranean area is intermediate between the southern part of the area and the other parts of Europe.

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[6] - Hunzelmann N, Baratli J, Asturias J. High prevalence of sensitization to plane allergens in Cologne. Allergy 2007;62(suppl. 83):171

Background: Platanus acerifolia pollen is an important contributor to allergic symptoms in South European and American cities, particularly in Southern Europe. To date published data only describe South-European cohorts, whereas little is known about the sensitization to plane pollen in Germany. Therefore we analyzed the frequency and pattern of sensitization to plane pollen in patients seen at the allergy outpatient clinic of the University Hospital of Cologne. Methods: 668 patients were tested by skin-prick tests (SPT). In addition, further analysis was carried out by determination of specific IgE-reactivity using the phadia ImmunoCAP system and reactivity to the major known recombinant allergens from P. Acerifolia i.e. Pla a 1, Pla a 2, plane profilin by EAST. In patients unreactive by EAST or CAP westernblot analysis was performed. Results: The prevalence of sensitization to P. acerifolia among 668 patients was 13,2% (88 patients). 7,5% (50) showed positive IgE-reactivity by SPT, 3,6% (24) by ImmunoCAP, 2,1% (14) to both. 75% of sensitized patients showed a total IgE > 100 kU/l. Furthermore, 35 patients positive by ImmunoCAP, were tested for specific IgE levels against recombinant Pla a 1, natural Pla a 2, and natural plane profilin. The test showed no clear concordance between ImmunoCAP and EAST, some of the sera had a positive CAP but no IgE-binding against purified allergens. Sera investigated by Western blotting against plane pollen extract demonstrated widespread IgE-reactivity and IgE-binding pattern. Conclusion: The results demonstrate a marked prevalence of plane sensitization in a central European city. Interestingly the complex IgE reactivity pattern found on sensitized patients parallels previous results described in southern Europe.

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[7] - Lopes Silva S, Rodrigues Alves R, Spínola Santos A, Pregal A, Mendes A, Pedro E, et al. Cypress Allergy in Portugal. J Allergy Clin Immunol 2005;115(2 suppl.):S238

RATIONALE: Since cypress allergy has been claimed as a major cause of winter rhinoconjuntivitis and asthma in the Mediterranean area, we wanted to evaluate if this extract should be included in our Unit‚s standard panel of allergens METHODS: We studied 105 sequential patients with respiratory allergy Prick tests to Cupressus sempervirens (Cs), Cupressus arizonica (Ca) and Juniperus ashei (Ja) were performed along with a standard panel of allergens including grass mix, parietaria, ragweed, plantain, olive, platanus, dust mite, cat and dog dander and fungi. Specific IgE (sIgE) was performed only in selected cases RESULTS: Among the 105 patients evaluated (65F/40M; mean age: 32.0±14.8 years), 45.7% had rhinitis, 1.9% asthma and 52.3% had both diseases. Cutaneous sensitization to cupressaceae (21,9%), was the fourth most prevalent pollen sensitization after grasses (49.5%), olive (24.8%) and parietaria (22.8%). Comparing patients sensitized to cupressaceae (C+) with non-sensitized (C-), no significant differences were found concerning sex or age distribution, clinical manifestations, total IgE or family history of atopy. Co-sensitization prevalences comparison only presented a significant statistic difference concerning Platanus (21.7% of C+ and 2.4% of C-, p<0.05). sIgE to Cs was performed in 16 patients with cutaneous sensitization and was positive in 5. Only 2 patients with cutaneous sensitization had winter related symptoms, both with indetectable sIgE CONCLUSION: Cypress cutaneous sensitization is frequent and the extract should be added to our standard panel of allergens. Specific IgE to Ca and Ja currently being performed in our population may help to establish cypress allergy real clinical relevance, still not clearly demonstrated

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[8] - Minciullo PL, Patafi M, Marotta G, Tigano V, Barresi L, Ferlazzo B, et al. Platanus acerifolia co-sensitivity in patients with parietaria judaica allergy and oral allergic syndrome. Allergy Clin Immunol Int 2005;17(Suppl. 1):250

Plane tree pollen allergy is a clinical disorder affecting particularly populations of the United States and Western Europe, but little is known about its relevant allergens. Pla a 1 and Pla a 2 are known as major allergens of Platanus acerifolia. Recently, a new allergen, called Pla a 3 has been characterized as major allergen in plane pollen allergic patients with concomitant allergy to vegetables and/or fruits in the Mediterranean area. It is a lipid transfer protein (LTP) that cross-reacts with food LTPs, in particular with lettuce, cherry and hazelnut LTP. Other previous studies described an association between the pollen of the Platanus tree and hazelnut, peach, apple, banana, peanut, celery, maize and chickpea. This association could be explained by the in vitro IgE crossreactivity detected. No data are present about a possible cross-reactivity between plane tree pollen and other plant or thee pollens. During the period between January 2004 and November 2004, 14 patients, referred to our Operative Unit for seasonal rhinitis and/or asthma symptoms, showed positivity to skin prick test to plane tree. All patients showed co-sensitivity to other pollens and in particular 13 out of 14 patients presented skin positivity to Parietaria judaica, the most important pollen in our area. Other important co-sensitivities were with Olea europea (10 patients) and grass pollen (7 patients). No monosensitivity to platanus was found. Therefore, platanus sensitivity in our area seems no to be so important and might be related to parietaria pollen. Moreover, 5 out of 14 patients with platanus sensitivity, referred an oral allergic syndrome (OAS) and showed skin positivity for vegetables and/or fruits. All of 5 patients were sensitive to peach, 4 to apple, 4 to hazelnut, 4 to almond, 4 to beet, 3 to peanut and 3 to walnut. Since in the Mediterranean area OAS occurs independently of an associated birch pollinosis and since platanus pollinosis in our area seems no to play an important role, it may be hypothesised that the skin positivity to platanus is related to fruit LTP in patients with OAS. The role of parietaria allergens in cross-reactivity with platanus and fruit LTP should be investigate.

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[9] - Mari A. Multiple pollen sensitization: a molecular approach to the diagnosis. Int Arch Allergy Immunol 2001;125:57-65

BACKGROUND: Sensitization to multiple pollen species is a frequent diagnostic event. Several allergenic molecules with a high level of homology have been identified in divergent pollen families and named panallergens. METHODS: We sought to define the criteria to evaluate the prevalence of the multiple pollen sensitization, to identify specific markers of this condition, and to correlate them with the underlying allergic disease. Patients presenting an allergic respiratory disease underwent skin testing with 23 pollens. Patients fulfilling predefined selection criteria were grouped and classified as having multiple pollen sensitization. Patients in each subgroup were tested for IgE to rBet v 2, rJun o 2, rBet v 1, rPhl p 5 and bromelain. Demographical, allergological and clinical data were recorded in the subgroup of patients with multiple pollen sensitization. RESULTS: Seventeen percent of the pollen-sensitized patients formed the multiple pollen-sensitized subgroup. These subjects were positive for most of the pollen species tested regardless of known exposure to them. None of the subjects sensitized to less than six pollen species were positive to panallergens, whereas 55% of the sera of the multiple pollen-sensitized group were positive to rBet v 2, and 15% to rJun o 2. IgE to rBet v 1 and rPhl p 5 were found positive in all the subgroups. Age, gender, bronchial asthma, oral allergy syndrome, skin test reactivity and previous specific immunotherapy differed significantly when these two subsets were considered. CONCLUSIONS: Allergy diagnosis based on allergenic molecules is crucial in the patient with multiple pollen sensitization. This condition appears to be determined by the sensitization to defined allergenic components (panallergens) rather than by pollen of multiple species as such. Detection of IgE to nonpanallergenic molecules allows to identify more relevant allergenic sources. Clinical aspects of the underlying allergic disease (e.g. asthma and oral allergy syndrome) seem to be differently related to IgE reactivity to panallergens

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[10] - Alcázar P, Cariñanos P, De Castro C, Guerra F, Moreno C, Domínguez-Vilches E, et al. Airborne plane-tree (Platanus hispanica) pollen distribution in the city of Córdoba, South-western Spain, and possible implications on pollen allergy. J Investig Allergol Clin Immunol 2004;14:238-243

Plane-trees are widely grown in Spain as ornaments. This taxon releases large amounts of pollen into the air and this pollen type seems to have proved implications on pollinosis. The aim of this study was to analyse airborne Platanus pollen content in the air of C6rdoba (South-western Spain) over the last 10 years, and its distribution in different areas of the city, as well as to consider possible implications regarding pollen allergies. Results revealed that Platanus pollen concentrations have been at their highest over the last 5 years. Within the city, the lowest pollen concentrations were in Northern areas of the city. Nevertheless, concentrations high enough to be of risk are reached over the whole city. All the patients studied presented symptoms during the Platanus pollen season. We can therefore conclude that plane pollen presents a risk for the whole city although symptoms are more intense and persistent in districts with a higher number of plane-trees. Platanus allergy is a particular problem for city-dwellers, where these trees are commonly used as ornamentals.

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[11] - Cosmes Martin PM, Moreno Ancillo A, Dominguez Noche C, Gutierrez Vivas A, Belmonte Soler J, Roure Nolla JM. [Sensitization to Castanea sativa pollen and pollinosis in northern Extremadura (Spain)]. Allergol Immunopathol (Madr) 2005;33:145-150

BACKGROUND: Castanea sativa pollen allergy has generally been considered to be uncommon and clinically insignificant. In our geographical area (Plasencia, Caceres, Spain) Castanea sativa pollen is a major pollen. OBJECTIVE: To determine the atmospheric fluctuations and prevalence of patients sensitized to Castanea pollen in our region and to compare this sensitization with sensitizations to other pollens. METHODS: Patients with respiratory symptoms attending our outpatient clinic for the first time in 2003 were studied. The patients underwent skin prick tests with commercial extracts of a battery of inhalants including Castanea sativa pollen. Serologic specific IgE to Castanea sativa pollen was determined using the CAP system (Pharmacia and Upjohn, Uppsala, Sweden). Airborne pollen counts in our city were obtained using Cour collection apparatus over a 4-year period (2000 to 2003). RESULTS: The most predominant pollens detected were (mean of the maximal weekly concentrations over 4 years in pollen grains/m3): Quercus 968, Poacea 660, Olea 325, Platanus 229, Pinus 126, Cupresaceae 117, Plantago 109, Alnus 41, Populus 40, Castanea 32. We studied 346 patients (mean age: 24.1 years). In 210 patients with a diagnosis of pollinosis, the percentages of sensitization were: Dactylis glomerata 80.4%, Olea europea 71.9%, Fraxinus excelsior 68%, Plantago lanceolata 62.8%, Chenopodium album 60.9%, Robinia pseudoacacia 49%, Artemisia vulgaris 43.8%, Platanus acerifolia 36.6%, Parietaria judaica 36.1%, Populus nigra 32.3%, Betula alba 27.6%, Quercus ilex 21.4%, Alnus glutinosa 20.9%, Cupressus arizonica 7.6% and Castanea sativa 7.1%. Fifteen patients were sensitized to Castanea sativa and 14 had seasonal rhinoconjunctivitis and asthma. Ten patients had serum specific IgE to Castanea pollen (maximum value: 17.4 Ku/l). Castanea pollen is present in our area in large amounts from the 23rd to the 28th weeks of the year, with a peak pollen count in the 25th week. CONCLUSIONS: The most important allergenic pollens in northern Extremadura were Poaceae, Olea europaea and Plantago sp. The prevalence of sensitization to Castanea sativa pollen was very low (7.1%). Most sensitized patients had asthma and polysensitization. Castanea sativa pollen is not a major cause of pollinosis in our area.

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[12] - Varela S, Subiza J, Subiza JL, Rodriguez R, Garcia B, Jerez M, et al. Platanus pollen as an important cause of pollinosis. J Allergy Clin Immunol 1997;100:748-754

OBJECTIVE: The existence of Platanus pollinosis is not generally accepted despite the production of very large quantities of airborne Platanus pollen in many cities of the United States and Europe. The aim of this study was to investigate if Platanus pollen really contributes to the symptoms of the patients with pollinosis in the Madrid area. METHODS: We carried out systematic skin prick testing with Platanus pollen extract on 47 patients seen in our allergy center with spring-summer pollinosis symptoms. Each patient maintained symptom score diaries before, during, and after the Platanus pollination season. The average symptom scores were calculated and compared with the Platanus pollen counts. Measurements of specific IgE by ELISA and immunoblotting also were performed in each patient. RESULTS: The Platanus skin tests were positive in 33 of the 39 patients first seen with seasonal symptoms during Platanus pollen season and only in three of the eight patients without symptoms during Platanus exposure (Fisher's exact test; p < 0.05). Twenty-two of the 33 Platanus-positive skin test patients also had a positive ELISA result. Furthermore, the average 24-hour rhinitis symptom scores of the 39 patients first seen with seasonal symptoms during March through April showed significant correlation with Platanus pollen counts (r(s) = 0.57, p < 0.05). The immunoblot results suggest that a 17 kd pollen protein could be a major allergen in patients with Platanus pollinosis. CONCLUSIONS: Platanus pollen is an important cause of pollinosis in Madrid. A protein with a molecular weight of 17 kd appeared to be its major allergen.

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[13] - Enrique E, Cisteró-Bahíma A, Bartolomé B, Alonso R, San Miguel-Moncín MM, Bartra J, et al. Platanus acerifolia pollinosis and food allergy. Allergy 2002;57:351-356

Background:In Mediterranean areas, oral allergy syndrome (OAS) occurs independently of an associated birch pollinosis; moreover, on occasions it presents with no other associated pollinosis. The aim of this study was to assess the possible association of OAS with Platanus acerifolia pollinosis. Methods:We evaluated consecutive patients seen for pollinosis in an allergy department. Seven hundred and twenty patients were selected on the basis of seasonal or perennial rhinitis, or asthma, or both. Respiratory and food allergies were studied in all patients. Clinical history was recorded and examinations and skin prick tests were performed with a battery of available common inhalant allergens and plant-derived food allergens. Specific IgE levels to P.acerifolia pollen extract and food allergens tested were measured. Molecular masses of the IgE-binding proteins and cross-reactivity among the P. acerifolia pollen and different food extracts were also determined. Results:Of the 720 patients evaluated, 61 (8.48%) were sensitized to P.acerifolia pollen. Food allergy was observed in 32 (52.45%) of the 61 patients sensitized to P.acerifolia pollen. Food allergens most frequently implicated were hazelnuts, peach, apple, peanuts, maize, chickpea and lettuce. Enzyme allergosorbent (EAST)-inhibition showed high inhibition values when P.acerifolia pollen extract was used as free phase. On the contrary low inhibition was observed when plant-derived food allergens were used as free phase and P.acerifolia pollen extract as solid phase. Conclusions:Cross-reactivity was observed among P.acerifolia pollen and plant-derived foods. OAS in these patients may have been caused by primary respiratory sensitization. [= AAAAI 58th Annual Meeting, New York, 1-6 March, 2002, Poster n°936]

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[14] - Tella R, Dall'Aglio M, San Miguel-Moncin M, Alonso R, Bartra J, Fernandez-Parra B, et al. Relationship between Platanus acerifolia pollinosis and food allergy in Barcelona Area. AAAAI 60th Annual Meeting, San Francisco, 19-23 March 2004, Poster n°480

Rationale Association of certain pollinosis with plant-derived food allergy has been demonstrated in several studies. The aim of this study was to analyze the clinical features and the prevalence of food allergy comparing patients sensitized to different pollen allergens (Platanus acerifolia versus other prevalent pollen allergies in our region). Methods From 6739 consecutive patients attending an allergy outpatient clinic during the last 10 years, 1477 suffered pollen allergies. Seventy-three (4.9%) patients suffered asthma, 919 rhinoconjunctivitis (62.2%) and 484 (32.8%) asthma and rhinoconjunctivitis. Patients were divided in a group sensitized to Platanus pollen (n=218, 14.7%), sensitized to grass (n=601, 40.6%) and sensitized to both pollens (n=122, 8.2%). We evaluated food allergy by clinical history, skin prick test +/ˆ challenge tests if proceeds in every group, and we compared which kind of food was implicated in each pollinosis. Results A 28.8% of Platanus sensitized patients suffered food allergy, while this fact only appeared in a 6.4% of the grass sensitized group (p<0.001). In the group of patients sensitized to both pollens, the prevalence of food allergy was similar to the Platanus sensitized group (27.8%). Regarding the clinical features of food allergy found in every group, systemic symptoms appeared in a 47.6% of Platanus group, a 38.9% of grass group and a 35.2% of patients from the group sensitized to both pollens. Conclusions Patients with pollinosis to Platanus could present more frequently food allergy compared with other pollinosis. Moreover, systemic reactions caused by food allergy are more frequent in patients sensitized to Platanus than in the other groups.

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[15] - Corominas M, Lleonart R, Martin C, De la Torre F. Profile of allergen pollen reactivity in patients sensitised to Olea pollen. Allergy 2008;63(suppl. 88):104

Background: Previous studies performed in our area (with low levels of Olea pollen) showed that 38% of patients with pollinosis were sensitised to Olea, although only 7% of them are monosensitised. The aim of this study is to evaluate sensitisation to olive pollen as a consequence of cross reactivity with other pollen allergens. We studied the sensitisation pollen profile of patients with positive skin prick test (SPT) to Olea and the cross reactivity with other pollen allergens. Patients and methods: Fourteen adult patients (8 males and 6 females) whith a positive SPT to Olea pollen were included. We analyzed their clinical history and SPT to common airborne allergens and allergens with a high LTP, profilin or polcalcin content. Specific serum IgE against to recombinant allergens Ole e1, Ole e 9, Salk1, Par j 1, Phl p 1, Phl p 5, Art v 1, Cup s 1, Bet v 1, Pru p 3, Mal d 4, Che a 3 (ALK-Abello) were quantified in the ADVIA Centaur platform. Results: Ten patients presented rhinitis and four asthma and rhinitis. According to SPT, 78% were positive to grasses, 64% to Parietaria, 57% to Salsola, 57% to Plantago, 50% to Platanus, 43% to Artemisia, 36% to Cupressus, 36% to peach, 21% to polcalcin, 14% to profilin and 14% to Betula. When we analyzed specific IgE against recombinant allergens, we found that 11 of the 14 patients (78%) were reactive to Ole e1 and none to Ole e 9. Only one patient had specific IgE to Che a 3 (polcalcin), and another to Che a 3 and to Mal d 4 (profilin). Ole e 1 specific IgE levels correlated with IgE levels of Phl p 1, Phl p 5, Pru p 3, Cup s 1, Che a 3, Mal d4, Pla l 1 and Bet v 1 (P<0.002). Conclusions: Most of the patients sensitised to Olea had specific IgE to Ole e 1 allergen. Profilin and polcalcin allergens are not responsible for polysensitisation present in these patients. In this group, sensitisation to Olea pollen corresponds to the great number of sensitisation present, but it is not a result of cross reactivity.

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[16] - EXPO: Determinacion de la prevalencia de sensibilizacion a polenes mediante tecnicas de diagnostico molecular. ALK-Abello Ed., 2009, 86pp.

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[17] - Goikoetxea M, Gastaminza G, Fernandez-Benitez M, Ferrer M, Sanz M. Microarray of allergenic component-resolved diagnosis in polysensitised patients. Allergy 2008;63(suppl. 88):42

Background: Component-resolved diagnosis (CRD) applied to allergy based in quantification of specific IgE against allergenic molecules emerges as a very useful tool for an acute diagnosis of sensitisations. The aim of this study was to analyze the sensitisation profile to allergenic compounds of polysensitised allergic patients diagnosed previously using standard techniques. Material and methods: In plasma from 79 atopic polysensitised patients defined as two or more sensitisations to different allergenic sources, specif IgE was measured semiquantitatively against 79 allergenic components (32 patients) and against 85 allergenic components (47 patients) using the microarray ISAC CRD 79 and 85, respectively (VBC-genomics, Phadia, Suecia). Results: 92% (73/79) of the patients were sensititized to any pollen allergen tested. The prevalence of sensitisation to any grass pollen allergen was 75% (59/79), to Betula v. Pollen allergens was 43% (34/79), to olive pollen allergens was 54% (43/79), to Platanus a. pollen allergens was 18% (14/79) and to Cupressus a. allergen was 39% (31/79). Some sensitisations are shared. All grass pollen sensitised patients showed specific IgE against Phl p 1 and 64% (38/59) of them showed also sensitisation to Phl p 5. In addition, 20% (12/59) of the patients sensitised to grass pollen major allergens (Phl p 1 and Phl p 5) were also sensitised to profillins. We also observed that 65% (28/ 43) of the patients sensitised to olive pollen allergens were sensitised to the profilin ole e 2 and 71% (24/34) of the patients sensitised to Betula v. pollen allergens were sensitised to the profillin Bet v 2. 44% (35/79) of the patients were sensitised to house dust mite, 14% (5/35) of these patients were also sensitised to all TPM tested in the microarray. From the 37% (29/79) of the patients sensitised to any LTP, most of them (23/79) were sensitised to peach LTP Pru p 3 whereas only 5 patients sensitised to Parietaria j. pollen (Par j 1 y Par j 2) LTPs. Only one patient was sensitised to all LTPs from the two different allergenic sources. In summary, we observed that 48% (38/79) of the polisensitised patients were sensitised to panallergens. Conclusion: CRD applied to allergy is a practical tool for polysensitised patients and could stablish sensitisation profile acutely. Application of specific IgE determination by microarrays technology could improve the diagnosis of allergy diseases and could modify therapeutical management in the future.

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[18] - Fernández-Rivas M, González-Mancebo E, Rodríguez-Pérez R, Benito C, Sánchez-Monge R, Salcedo G, et al. Clinically relevant peach allergy is related to peach lipid transfer protein, Pru p 3, in the Spanish population. J Allergy Clin Immunol 2003;112:789-795

BACKGROUND: Sensitization to peach and related Rosaceae fruits without clinical expression is commonly observed as the result of the extensive cross-reactivity of IgE antibodies directed toward lipid transfer proteins (LTPs), Bet v 1 homologues, profilins, and carbohydrate determinants . OBJECTIVE: We aimed to study whether there are any clinical or immunologic differences between patients allergic to peach and those who have a current clinically irrelevant sensitization to this fruit . METHODS: One hundred subjects with adverse reactions to peach were evaluated by medical history, skin prick tests with fresh peach and purified peach LTP (Pru p 3), and specific IgE determinations to peach, rBet v 1, and rBet v 2 (birch profilin). Clinical reactivity to peach was established by double-blind, placebo-controlled food challenges. The clinical characteristics and the in vivo and in vitro tests were compared between allergic and nonallergic patients . RESULTS: Peach allergy was confirmed in 76 patients and ruled out in 16; 2 patients dropped out, and the study was not conclusive in 6 individuals (placebo reactors). Pollen allergy was found in 76% of the allergic patients and in 100% of the nonallergic patients. Positive responses to Pru p 3, rBet v 1, and rBet v 2 were observed in 62%, 7%, and 34% of patients allergic to peach, respectively. The sensitization rate to Pru p 3 was significantly higher among subjects allergic than nonallergic to peach (62% vs 31%, P =.02). IgE responses to rBet v 2 were more frequent among subjects allergic to pollen, but no difference was observed in the presence or absence of peach allergy . CONCLUSIONS: Pru p 3 is the major allergen of peach in our population, and the IgE response to this allergen is related to the clinical expression of peach allergy. Sensitization to profilin is observed in those patients with an associated pollen allergy but does not appear to be related to the clinical reactivity to peach.

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[19] - Alemán A, Sastre J, Quirce S, de las Heras M, Carnés J, Fernández-Caldas E, et al. Allergy to kiwi: A double-blind, placebo-controlled food challenge study in patients from a birch-free area. J Allergy Clin Immunol 2004;113:543-550

Background Allergy to kiwi fruit is being increasingly reported, but it has never been evaluated by means of a double-blind, placebo-controlled food challenge (DBPCFC) study. Objective : We sought to assess kiwi allergy on the basis of a DBPCFC and identify the patterns of allergen recognition in sensitized patients from a birch-free area. Method s : Forty-three patients with allergy symptoms who were sensitized to kiwi were evaluated by means of clinical history, skin tests, IgE determinations, and DBPCFCs. The pattern of allergen recognition was assessed by means of IgE immunoblotting. Sequence analysis of IgE-binding bands was performed by using Edman degradation. Result s : DBPCFCs were performed in 33 patients; 4 patients had experienced severe anaphylaxis, and 6 patients declined informed consent. DBPCFC results were positive in 23 patients and negative in 10 patients. The most frequent clinical manifestation was oral allergy syndrome. Twenty-one percent of the patients were not allergic to pollen. Forty-six percent of patients experienced systemic symptoms, and this happened with higher frequency in patients not allergic to pollen (100%). Twenty-eight percent of the patients were sensitized to latex. The IgE-binding bands in kiwi extract more frequently recognized by patient sera were those of 30, 24, 66, and 12 kd, and they could not be associated with any pattern of kiwi-induced allergic reactions. Conclusion : The results provide evidence that kiwi allergy is not a homogeneous disorder because several clinical subgroups can be established. No definite allergen-recognition pattern was associated with the type of allergic reactions to kiwi. One of 5 patients with kiwi allergy was not allergic to pollen, and these patients had the highest risk of systemic reactions to kiwi.

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[20] - Figueroa J, Blanco C, Dumpiérrez AG, Almeida L, Ortega N, Castillo R, et al. Mustard allergy confirmed by double-blind placebo-controlled food challenges: clinical features and cross-reactivity with mugwort pollen and plant-derived foods. Allergy 2005;60:48-55

BACKGROUND: Mustard IgE-mediated allergy is supposed to be a rare cause of food allergy, and its clinical features and cross-reactivities have not been fully elucidated . METHODS: A prospective study was carried out, recruiting mustard allergic patients, and paired control subjects. A clinical questionnaire was administered, and skin-prick tests (SPT) with panels of aeroallergens and foods, serum extraction for in vitro tests and double-blind placebo-controlled food challenges (DBPCFC) were performed . RESULTS: Thirty-eight mainly adult patients, with 10.5% reporting systemic anaphylaxis, were included in the study [age (mean +/- SD): 21.9 +/- 8.6 years]. DBPCFC were performed in 24 patients, being positive in 14 cases (58.3%). Patients with positive outcome showed significantly greater mustard SPT than those with negative outcome (8.2 +/- 3.7 vs 5.3 +/- 2.4 mm, P <0.05), and the receiver-operating characteristic (ROC) curve analysis yielded a cut-off value for mustard commercial SPT of 8 mm, with a specificity of 90% (95% CI, 55.5-98.3), and a sensitivity of 50% (95% CI, 23.1-76.9). A significant association between mustard hypersensitivity and mugwort pollen sensitization was found (97.4% of patients), with partial cross-reactivity demonstrated by UniCAP System inhibition assays. All patients showed sensitization to other members of Brassicaceae family, and cross-reactivity among them was also confirmed. Moreover, significant associations with nut (97.4%), leguminous (94.7%), corn (78.9%), and Rosaceae fruit (89.5%) sensitizations were also shown. Around 40% of these food sensitizations were symptomatic, including food-dependent exercise-induced anaphylaxis in six patients . CONCLUSIONS: Mustard allergy is a not-uncommon disorder that can induce severe reactions. Significant associations with mugwort pollinosis and several plant-derived food allergies are demonstrated, suggesting a new mustard-mugwort allergy syndrome. A relationship between this syndrome and food-dependent exercise-induced anaphylaxis is also reported.

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[21] - Asturias JA, Ibarrola I, Eraso E, Arilla MC, Martinez A. The major Platanus acerifolia pollen allergen Pla a 1 has sequence homology to invertase inhibitors. Clin Exp Allergy 2003;33:978-985

BACKGROUND: Sycamores or plane trees are an important source of airborne allergens in many cities of the United States and Western Europe. Pla a 1 has been described as a major allergen from Platanus acerifolia (London plane tree) . OBJECTIVE: To clone and characterize the cDNA for Pla a 1 and to express the recombinant protein . METHODS: Pla a 1 was isolated by cationic exchange, gel filtration, and reverse-phase chromato-graphies. Pla a 1 cDNA was cloned by reverse transcription followed by polymerase chain reaction, using amino acid sequences from tryptic peptides of the allergen. The Pla a 1 encoding sequence has been subcloned into the pKN172 expression vector and expressed in Escherichia coli as a non-fusion protein. Purified recombinant protein has been tested for its IgE-binding capacity in immunoblot, immunoblot inhibition, and ELISA . RESULTS: Pla a 1 reacted with serum IgE from 35 of the 42 (83.3%) Platanus-allergic patients studied and represented 60% of the total IgE-binding capacity of the P. acerifolia pollen extract. The allergen displayed 43% sequence identity to a grape invertase inhibitor and showed a predicted secondary structure characteristic of all-alpha proteins. Serological analysis revealed that both natural and recombinant forms of Pla a 1 displayed similar IgE-binding capacity . CONCLUSIONS: Pla a 1 belongs to a new class of allergens related to proteinaceous invertase inhibitors. Recombinant Pla a 1 binds IgE in vitro like its natural counterpart and, therefore, it can be useful for specific diagnosis and structural studies.

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[22] - Asturias JA, Ibarrola I, Amat P, Tella R, Malet A, Cisteró-Bahíma A, et al. Purified allergens vs. complete extract in the diagnosis of plane tree pollen allergy. Clin Exp allergy 2006;36:1505-1512

BACKGROUND: Plane tree pollen allergy is a clinical disorder affecting human population in cities of Europe, North America, South Africa, and Australia . OBJECTIVE: To compare IgE-reactivity of the natural and recombinant forms of two major plane allergens, Pla a 1 and Pla a 2, with the reactivity of Platanus acerifolia pollen extract . METHODS: Forty-seven patients with P. acerifolia allergy, 15 of them monosensitized, and 24 control subjects were included in the study. Natural Pla a 1 and Pla a 2 were purified by standard chromatographic methods and recombinant proteins were expressed in Escherichia coli. Skin prick test and determination of specific IgE were performed with commercial P. acerifolia extract and natural and recombinant purified allergens . RESULTS: Pla a 1 and Pla a 2 were responsible for 79% of the IgE-binding capacity against P. acerifolia pollen extract. A high correlation has been found between the IgE response to nPla a 1 (R = 0.80; P < 0.001) or nPla a 2 (R = 0.79; P < 0.001) vs. P. acerifolia extract as well as between natural and recombinant Pla a 1 (R = 0.89; P < 0.001). Skin testing showed no significant differences between extract and nPla a 2, whereas a higher reactivity was found with nPla a 1. In contrast, rPla a 1 revealed markedly reduced sensitivity in comparison with extract by skin prick test and specific IgE. The sensitivity of the mix Pla a 1+Pla a 2 was 100% and 87.5% for monosensitized and polysensitized patients, respectively, with no false-positive reactions detected. Conclusion Pla a 1 and Pla 2 are sufficient for a reliable diagnosis of P. acerifolia in most patients and induce comparable skin test reactivity as a whole extract.

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[23] - Asturias JA, Ibarrola I, Eraso E, Arilla MC, Martinez A. The major Platanus acerifolia pollen allergen Pla a 1 has sequence homology to invertase inhibitors. Clin Exp Allergy 2003;33:978-985

BACKGROUND: Sycamores or plane trees are an important source of airborne allergens in many cities of the United States and Western Europe. Pla a 1 has been described as a major allergen from Platanus acerifolia (London plane tree) . OBJECTIVE: To clone and characterize the cDNA for Pla a 1 and to express the recombinant protein . METHODS: Pla a 1 was isolated by cationic exchange, gel filtration, and reverse-phase chromato-graphies. Pla a 1 cDNA was cloned by reverse transcription followed by polymerase chain reaction, using amino acid sequences from tryptic peptides of the allergen. The Pla a 1 encoding sequence has been subcloned into the pKN172 expression vector and expressed in Escherichia coli as a non-fusion protein. Purified recombinant protein has been tested for its IgE-binding capacity in immunoblot, immunoblot inhibition, and ELISA . RESULTS: Pla a 1 reacted with serum IgE from 35 of the 42 (83.3%) Platanus-allergic patients studied and represented 60% of the total IgE-binding capacity of the P. acerifolia pollen extract. The allergen displayed 43% sequence identity to a grape invertase inhibitor and showed a predicted secondary structure characteristic of all-alpha proteins. Serological analysis revealed that both natural and recombinant forms of Pla a 1 displayed similar IgE-binding capacity . CONCLUSIONS: Pla a 1 belongs to a new class of allergens related to proteinaceous invertase inhibitors. Recombinant Pla a 1 binds IgE in vitro like its natural counterpart and, therefore, it can be useful for specific diagnosis and structural studies.

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[24] - Palazzo P, Scala E, Bernardi ML, Mari A. Immunochemical and diagnostic impact of CCD-IgE reactivity on microarrayed allergens. Allergy 2007;62(suppl. 83):152-153

Background: The asparagine-linked carbohydrate moieties of plant and insect glycoproteins are the structural basis of what is known as IgE cross-reactive carbohydrate determinants (CCD). About 20% or more of allergic patients have been reported to produce specific anti-glycan IgE. CCD do not appear to cause clinical symptoms. We sought to establish the interference of anti-CCD IgE on a proteomic microarray system having both native and recombinant spotted allergens, including several glycoproteins. Methods: A population of 6155 subjects suspected for having an IgE-mediated disease has been screened using the multiplexed ISAC system (ISAC CRD 79b, VBC-Genomics, Austria). A serum was considered positive for CCD reactivity when a positive IgE reaction to Ana c 2 (bromelain) was detected. Twelve CCD-IgE positive subjects, having high IgE titers against Ana c 2, were tested on two more singleplex systems: CAP (Phadia, Sweden) and Immulite (Siemens, CA). IgE reactivity was detected for known glycoproteins rarely acting as true allergens, Ana c 2 and horseradish peroxidase, and to a purified glycopeptide from bromelain (MUXF3). A CCD-IgE adsorption was performed by using immobilized MUXF3 on CAP, then tested on ISAC system. Results: 4.2 % of the microarray tested population resulted positive for Ana c 2. A cluster reactivity was recorded for known glycoproteins, Ana c 2, Api m 1, but also for allergens never described for their glycan side chain IgE reactivity: Hor v 17, and Pla a 2. Allergens with a possible IgE reactivity for either CCD or protein epitopes (Lol p 1, Ole e 1, Cup a 1, Art v 1) were differently involved in this cluster reactivity. All the 12 tested subjects were recorded positive for tested glycoproteins and glycans on both in vitro systems. IgE adsorption of the tested serum exerted a complete inhibition of IgE reactivity to Ana c 2, Hor v 17, Api m 1, Pla a 2, Cup a 1, and Ole e 1. The lack of inhibition of other glycoallergens was scored as a prevalent IgE protein epitope reactivity. Conclusions: CCD-IgE are detected on microarrayed glycosilated allergens. MUXF3 from bromelain represent a suitable marker for CCD-IgE reactivity, and could be spotted on microarray system to detect specific CCD-IgE. To overcome the problem of CCD-IgE reactivity non-glycosylated recombinant glycoallergens could be used. At the moment we suggest to use a two step approach by adsorption or inhibition with MUXF3 glycopeptide or a highly purified glycoprotein.

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[25] - Asturias JA, Ibarrola I, Amat P, Tella R, Malet A, Cisteró-Bahíma A, et al. Purified allergens vs. complete extract in the diagnosis of plane tree pollen allergy. Clin Exp allergy 2006;36:1505-1512

BACKGROUND: Plane tree pollen allergy is a clinical disorder affecting human population in cities of Europe, North America, South Africa, and Australia . OBJECTIVE: To compare IgE-reactivity of the natural and recombinant forms of two major plane allergens, Pla a 1 and Pla a 2, with the reactivity of Platanus acerifolia pollen extract . METHODS: Forty-seven patients with P. acerifolia allergy, 15 of them monosensitized, and 24 control subjects were included in the study. Natural Pla a 1 and Pla a 2 were purified by standard chromatographic methods and recombinant proteins were expressed in Escherichia coli. Skin prick test and determination of specific IgE were performed with commercial P. acerifolia extract and natural and recombinant purified allergens . RESULTS: Pla a 1 and Pla a 2 were responsible for 79% of the IgE-binding capacity against P. acerifolia pollen extract. A high correlation has been found between the IgE response to nPla a 1 (R = 0.80; P < 0.001) or nPla a 2 (R = 0.79; P < 0.001) vs. P. acerifolia extract as well as between natural and recombinant Pla a 1 (R = 0.89; P < 0.001). Skin testing showed no significant differences between extract and nPla a 2, whereas a higher reactivity was found with nPla a 1. In contrast, rPla a 1 revealed markedly reduced sensitivity in comparison with extract by skin prick test and specific IgE. The sensitivity of the mix Pla a 1+Pla a 2 was 100% and 87.5% for monosensitized and polysensitized patients, respectively, with no false-positive reactions detected. Conclusion Pla a 1 and Pla 2 are sufficient for a reliable diagnosis of P. acerifolia in most patients and induce comparable skin test reactivity as a whole extract.

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[26] - Asturias JA, Ibarrola I, Amat P, Tella R, Malet A, Cisteró-Bahíma A, et al. Purified allergens vs. complete extract in the diagnosis of plane tree pollen allergy. Clin Exp allergy 2006;36:1505-1512

BACKGROUND: Plane tree pollen allergy is a clinical disorder affecting human population in cities of Europe, North America, South Africa, and Australia . OBJECTIVE: To compare IgE-reactivity of the natural and recombinant forms of two major plane allergens, Pla a 1 and Pla a 2, with the reactivity of Platanus acerifolia pollen extract . METHODS: Forty-seven patients with P. acerifolia allergy, 15 of them monosensitized, and 24 control subjects were included in the study. Natural Pla a 1 and Pla a 2 were purified by standard chromatographic methods and recombinant proteins were expressed in Escherichia coli. Skin prick test and determination of specific IgE were performed with commercial P. acerifolia extract and natural and recombinant purified allergens . RESULTS: Pla a 1 and Pla a 2 were responsible for 79% of the IgE-binding capacity against P. acerifolia pollen extract. A high correlation has been found between the IgE response to nPla a 1 (R = 0.80; P < 0.001) or nPla a 2 (R = 0.79; P < 0.001) vs. P. acerifolia extract as well as between natural and recombinant Pla a 1 (R = 0.89; P < 0.001). Skin testing showed no significant differences between extract and nPla a 2, whereas a higher reactivity was found with nPla a 1. In contrast, rPla a 1 revealed markedly reduced sensitivity in comparison with extract by skin prick test and specific IgE. The sensitivity of the mix Pla a 1+Pla a 2 was 100% and 87.5% for monosensitized and polysensitized patients, respectively, with no false-positive reactions detected. Conclusion Pla a 1 and Pla 2 are sufficient for a reliable diagnosis of P. acerifolia in most patients and induce comparable skin test reactivity as a whole extract.

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[27] - Ibarrola I, Arilla MC, Martinez A, Asturias JA. Identification of a polygalacturonase as a major allergen (Pla a 2) from Platanus acerifolia pollen. J Allergy Clin Immunol 2004;113:1185-1191

Background Planetree pollen allergy is a clinical disorder affecting human populations in cities of the United States and Western Europe, but little is known about its relevant allergens. Objective : We sought to purify, characterize, and clone the 43-kd allergen from Platanus acerifolia. Method s : P acerifolia pollen extract was fractionated by using ion-exchange and gel-permeation chromatography. Analyses were carried out by using ELISA, SDS-PAGE, isoelectrofocusing, and immunoblotting. Partial amino acid sequence was obtained by means of Edman sequencing of cyanogen bromideˆdigested peptides. Specific cDNA was cloned by using reverse transcription, followed by PCR, with amino acid sequences from peptides of the allergen. Result s : The allergen isolated from P acerifolia pollen, Pla a 2, is a glycoprotein with an observed molecular mass of 43 kd and an isoelectric point value of 9.3. It is involved in the allergic responses of 84% of patients with planetree-induced pollinosis and represented 52% of the total IgE-binding capacity of the P acerifolia extract. Pla a 2 displays polygalacturonase (PG) activity, being the first PG with functional enzyme activity from an angiosperm plant pollen described as an allergen. The cDNA allergen sequence codified for a 372-residue protein with 56% and 42% sequence identity to PGs from pollen and fruits, respectively. Western blot analysis showed that Pla a 2 is present in pollen and stems and has IgG cross-reactivity with a PG from tomato and pectate lyases from Cupressaceae pollen. Conclusion : Pla a 2, a major allergen of P acerifolia pollen with PG activity has been purified, characterized, and cloned.

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[28] - Lauer I, San Miguel-Moncín M, Hartz C, Cistero-Bahima A, Conti A, Vieths S, et al. The lipid transfer protein (LTP) from Platanus acerifolia pollen, Pla a 3, is a major allergen and cross-reacts with food LTPs. EAACI 23th Congress, Amsterdam, 12-16 June, 2004, Poster n°1001

Background: Plane trees such as Platanus acerifolia are an important source of airborne allergens in many cities of Europe. Until now, only two major allergens of plane pollen, Pla a 1 and Pla a 2, have been characterised. In the Mediterranean area food-allergic patients are frequently sensitised to non-specific LTPs (nsLTPs). Sensitisation to food LTPs is not related to birch pollen allergy, and patients suffer from more severe reactions, in addition to OAS. An association between plane tree pollen allergy and plant food allergy was described, but the cross-reacting allergens have not been identified. The aim of this study was identification and characterisation of the nsLTP from plane pollen, and to investigate its immunological relationship to food LTPs from lettuce, cherry and hazelnut. Methods: Plane pollen extract was fractionated by ion exchange chromatography. Plane pollen LTP was identified by N-terminal amino acid sequence analysis. The prevalence of IgE reactivity to plane LTP was determined by immunoblotting using 45 Spanish plane pollen allergic patients with various plant food allergies and 14 plane pollen sensitised patients from Germany. In addition, sensitisation to LTPs from nuts, fruits and vegetables and their cross-reactivity to plane LTP was evaluated by immunoblotting and IgE inhibition experiments. Results: A 10 kDa IgE-reactive protein from plane pollen was partially purified and identified by N-terminal amino acid sequencing as nsLTP (Pla a 3). The N-terminal sequence of Pla a 3 showed the highest aa-ID of 58% with that of Lac s 1 (lettuce LTP). Pla a 3 was cross-reactive with food LTPs. Pla a 3 showed a prevalence of IgE reactivity of 66.6% (30/45) in Spanish plane pollen allergic patients. In contrast, Pla a 3 sensitisation was not relevant in plane pollen sensitised patients from Germany. We observed only partial correlation between sensitisation to plane pollen LTP and LTPs from lettuce, cherry and hazelnut. On the one hand, there were Spanish patients sensitised to food LTPs without plane pollen allergy, on the other hand, food LTP allergic patients without sensitisation to Pla a 3 were found. Conclusions: We have characterised the plane LTP as major allergen in plane pollen allergic patients with concomitant allergy to plant-derived foods recruited in the Mediterranean area. In Spanish patients sensitisation to food LTPs is not associated exclusively to the plane pollen LTP.

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[29] - Asturias JA, Ibarrola I, Amat P, Tella R, Malet A, Cisteró-Bahíma A, et al. Purified allergens vs. complete extract in the diagnosis of plane tree pollen allergy. Clin Exp allergy 2006;36:1505-1512

BACKGROUND: Plane tree pollen allergy is a clinical disorder affecting human population in cities of Europe, North America, South Africa, and Australia . OBJECTIVE: To compare IgE-reactivity of the natural and recombinant forms of two major plane allergens, Pla a 1 and Pla a 2, with the reactivity of Platanus acerifolia pollen extract . METHODS: Forty-seven patients with P. acerifolia allergy, 15 of them monosensitized, and 24 control subjects were included in the study. Natural Pla a 1 and Pla a 2 were purified by standard chromatographic methods and recombinant proteins were expressed in Escherichia coli. Skin prick test and determination of specific IgE were performed with commercial P. acerifolia extract and natural and recombinant purified allergens . RESULTS: Pla a 1 and Pla a 2 were responsible for 79% of the IgE-binding capacity against P. acerifolia pollen extract. A high correlation has been found between the IgE response to nPla a 1 (R = 0.80; P < 0.001) or nPla a 2 (R = 0.79; P < 0.001) vs. P. acerifolia extract as well as between natural and recombinant Pla a 1 (R = 0.89; P < 0.001). Skin testing showed no significant differences between extract and nPla a 2, whereas a higher reactivity was found with nPla a 1. In contrast, rPla a 1 revealed markedly reduced sensitivity in comparison with extract by skin prick test and specific IgE. The sensitivity of the mix Pla a 1+Pla a 2 was 100% and 87.5% for monosensitized and polysensitized patients, respectively, with no false-positive reactions detected. Conclusion Pla a 1 and Pla 2 are sufficient for a reliable diagnosis of P. acerifolia in most patients and induce comparable skin test reactivity as a whole extract.

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[30] - Lauer I, San Miguel-Moncín M, Hartz C, Cistero-Bahima A, Conti A, Vieths S, et al. The lipid transfer protein (LTP) from Platanus acerifolia pollen, Pla a 3, is a major allergen and cross-reacts with food LTPs. EAACI 23th Congress, Amsterdam, 12-16 June, 2004, Poster n°1001

Background: Plane trees such as Platanus acerifolia are an important source of airborne allergens in many cities of Europe. Until now, only two major allergens of plane pollen, Pla a 1 and Pla a 2, have been characterised. In the Mediterranean area food-allergic patients are frequently sensitised to non-specific LTPs (nsLTPs). Sensitisation to food LTPs is not related to birch pollen allergy, and patients suffer from more severe reactions, in addition to OAS. An association between plane tree pollen allergy and plant food allergy was described, but the cross-reacting allergens have not been identified. The aim of this study was identification and characterisation of the nsLTP from plane pollen, and to investigate its immunological relationship to food LTPs from lettuce, cherry and hazelnut. Methods: Plane pollen extract was fractionated by ion exchange chromatography. Plane pollen LTP was identified by N-terminal amino acid sequence analysis. The prevalence of IgE reactivity to plane LTP was determined by immunoblotting using 45 Spanish plane pollen allergic patients with various plant food allergies and 14 plane pollen sensitised patients from Germany. In addition, sensitisation to LTPs from nuts, fruits and vegetables and their cross-reactivity to plane LTP was evaluated by immunoblotting and IgE inhibition experiments. Results: A 10 kDa IgE-reactive protein from plane pollen was partially purified and identified by N-terminal amino acid sequencing as nsLTP (Pla a 3). The N-terminal sequence of Pla a 3 showed the highest aa-ID of 58% with that of Lac s 1 (lettuce LTP). Pla a 3 was cross-reactive with food LTPs. Pla a 3 showed a prevalence of IgE reactivity of 66.6% (30/45) in Spanish plane pollen allergic patients. In contrast, Pla a 3 sensitisation was not relevant in plane pollen sensitised patients from Germany. We observed only partial correlation between sensitisation to plane pollen LTP and LTPs from lettuce, cherry and hazelnut. On the one hand, there were Spanish patients sensitised to food LTPs without plane pollen allergy, on the other hand, food LTP allergic patients without sensitisation to Pla a 3 were found. Conclusions: We have characterised the plane LTP as major allergen in plane pollen allergic patients with concomitant allergy to plant-derived foods recruited in the Mediterranean area. In Spanish patients sensitisation to food LTPs is not associated exclusively to the plane pollen LTP.

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[31] - Asturias JA, Ibarrola I, Amat P, Tella R, Malet A, Cisteró-Bahíma A, et al. Purified allergens vs. complete extract in the diagnosis of plane tree pollen allergy. Clin Exp allergy 2006;36:1505-1512

BACKGROUND: Plane tree pollen allergy is a clinical disorder affecting human population in cities of Europe, North America, South Africa, and Australia . OBJECTIVE: To compare IgE-reactivity of the natural and recombinant forms of two major plane allergens, Pla a 1 and Pla a 2, with the reactivity of Platanus acerifolia pollen extract . METHODS: Forty-seven patients with P. acerifolia allergy, 15 of them monosensitized, and 24 control subjects were included in the study. Natural Pla a 1 and Pla a 2 were purified by standard chromatographic methods and recombinant proteins were expressed in Escherichia coli. Skin prick test and determination of specific IgE were performed with commercial P. acerifolia extract and natural and recombinant purified allergens . RESULTS: Pla a 1 and Pla a 2 were responsible for 79% of the IgE-binding capacity against P. acerifolia pollen extract. A high correlation has been found between the IgE response to nPla a 1 (R = 0.80; P < 0.001) or nPla a 2 (R = 0.79; P < 0.001) vs. P. acerifolia extract as well as between natural and recombinant Pla a 1 (R = 0.89; P < 0.001). Skin testing showed no significant differences between extract and nPla a 2, whereas a higher reactivity was found with nPla a 1. In contrast, rPla a 1 revealed markedly reduced sensitivity in comparison with extract by skin prick test and specific IgE. The sensitivity of the mix Pla a 1+Pla a 2 was 100% and 87.5% for monosensitized and polysensitized patients, respectively, with no false-positive reactions detected. Conclusion Pla a 1 and Pla 2 are sufficient for a reliable diagnosis of P. acerifolia in most patients and induce comparable skin test reactivity as a whole extract.

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[32] - Leduc-Brodard V, Inacio F, Jaquinod M, Forest E, David B, Peltre G. Characterization of Dac g 4, a major basic allergen from Dactylis glomerata pollen. J Allergy Clin Immunol 1996;98:1065-1072

Monoclonal antibodies were produced against Dac g 4, a purified major basic allergen from Dactylis glomerata pollen. Their ability to be used for immunopurification of Dac g 4 was studied on a BIAcore apparatus (Pharmacia). The allergen was purified by affinity chromatography with one monoclonal antibody. Its precise molecular mass, 59,185 +/- 30 d, was determined by mass spectrometry. Its isoelectric point is 10.4. Sodium dodecylsulfate-polyacrylamide gel electrophoresis and immunoblotting showed that Dac g 4-related proteins of similar molecular mass were detected in the majority of allergenic grass pollen species. By double-site ELISAs, we have estimated that Dac g 4 represents about 6% of the total proteins from a water-soluble extract. One monoclonal antibody (mAb H) recognized a 60 kd cross-reactive protein in other grass pollens, though none in any of the tree or weed pollens tested. Inhibition studies of IgE antibody binding to Dac g 4 with pollen extracts confirmed the presence of cross-reactive allergens in Secale cereale, Lolium perenne, Festuca elatior, Holcus lanatus, Bromus arvensis, Poa pratense, Hordeum sativum, and Phleum pratense.

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[33] - Arilla MC, Ibarrola I, Garcia R, de la Hoz B, Martinez A, Asturias JA. Quantification of the Major Allergen from Cypress (Cupressus arizonica) Pollen, Cup a 1, by Monoclonal Antibody-Based ELISA. Int Arch Allergy Immunol 2004;134:10-16

BACKGROUND: Cypress pollen allergy is an important cause of rhinoconjunctivitis and asthma in Mediterranean countries. Cypress allergenic extracts are difficult to produce since they have low protein and high carbohydrate content, thus accurate standardization of them is essential to guarantee their quality. The aim of this study is to develop a sandwich ELISA for the quantification of Cup a 1, the major allergen of cypress (Cupressus arizonica) pollen extract . METHODS: Monoclonal antibodies directed to purified Cup a 1 were produced. Two of them (9C7 as capture antibody and 3D2 as the tracer) were selected to develop a quantitative sandwich ELISA. This ELISA was subsequently evaluated and compared with other techniques . RESULTS: The described ELISA is very sensitive with a detection limit of 8.7 ng/ml and a practical working range of 62.5-1,000 ng/ml. The assay is also highly reproducible with intra-assay and interassay coefficients of variation of less than 10%. The purified Cup a 1, used as standard, presents pectate lyase enzymatic activity. The assay also detected Cup a 1-like proteins in pollen from other Cupressaceae. A good correlation was obtained between Cup a 1 content of 12 C. arizonica pollen extracts and their IgE-binding activity . CONCLUSIONS: The described Cup a 1 ELISA is sensitive, specific and reproducible and can be used for the quantification of Cup a 1 in C. arizonica and other related pollen extracts. It also provides a reliable indication of the allergenic activity of the whole cypress pollen extract.

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[34] - Arilla MC, Ibarrola I, Mir A, Monteseirin J, Conde J, Martinez A, et al. Development of a Sandwich-Type ELISA for Measuring Pla a 1, the Major Allergen of Platanus acerifolia Pollen. Int Arch Allergy Immunol 2005;138:127-133

Abstract Background:Platanus acerifolia is an important cause of pollinosis in Western European cities. Pla a 1, a nonglycosylated 18-kDa protein with a prevalence of 80%, is a major allergen in P. acerifolia pollen extracts. Our aim was to develop a Pla a 1-specific ELISA to quantify this protein in allergenic extracts and preparations for clinical use. Methods: Pla a 1 was purified by cation exchange at pH 7.0, gel filtration, and anion exchange chromatography at pH 10.0. Monoclonal (mAb) and polyclonal antibodies were obtained by immunizing mice and rabbits with nPla a 1. One (5C1) of the 13 mAb obtained was used as capture antibody at 5 mug/ml and biotin-labeled specific polyclonal antiserum at 0.63 mug/ml served for detection. Results: The prevalence of Pla a 1-specific IgE to purified Pla a 1 among 47 P. acerifolia-allergic patients was 79%. The Pla a 1-ELISA developed has a linear range of 3-25 ng/ml, high sensitivity with a detection limit of 0.5 ng/ml and is highly specific as none of the 24 pollen, mite, mol d, and plant food extracts tested gave positive results. The assay could quantify Pla a 1-like proteins in other planetree pollen extracts. A good correlation was obtained between Pla a 1 content of 11 P. acerifolia pollen extracts (average content 0.69% of the total protein) and their IgE-binding activity. Conclusions: The described two-site sandwich ELISA to measure Pla a 1 is useful for standardization of planetree pollen extracts intended for clinical use.

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[35] - Pazouki N, Sankian M, Nejadsattari T, Khavari-Nejad R, Varasteh AR. Oriental plane pollen allergy: Identification of allergens and cross-reactivity between relevant species. Allergy Asthma Proc 2008;29:622-628

Pollen from different tree and grass species represent the greatest inducers of type I allergy worldwide. Oriental plane trees, as Platanus orientalis, are an important source of airborne allergens in cities of the southwest Asia and southeast Europe. This study was aimed to identify relevant allergens of P. orientalis pollen and to ascertain whether P. orientalis allergens have cross-reactivity with related plane trees, such as Platanus acerifolia and Platanus occidentalis pollen components. Nineteen patients with a clinical history of reaction to P. orientalis pollen and a positive skin-prick test (SPT) to P. orientalis pollen extract were included in this study. Identification of IgE-binding proteins in Platanus pollen extracts was elucidated by immunoblotting using sera from P. orientalis pollen-sensitive patients. Cross-reactivity studies among P. orientalis allergens and relevant species was evaluated by immunoblot-inhibition and ELISA inhibition assays. All the patients were polysensitive and exhibited positive SPT to grass and tree pollen allergens. The IgE-binding pattern of P. orientalis pollen extract was well defined. The inhibition experiments showed that the capacity of P. occidentalis was greater than P. acerifolia pollen extract in preventing the reactivity of immobilized P. orientalis pollen extract with the IgE antibodies of patients. The 28-kDa molecule was the most frequent allergen among nine IgE-binding proteins of P. orientalis pollen. The IgE-reactive components of P. orientalis pollen showed a higher level of cross-reactivity with P. occidentalis pollen in comparison with P. acerifolia pollen.

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[36] - Pazouki N, Sankian M, Leung PT, Nejadsattari T, Khavari-Nejad RA, Varasteh AR. Identification of cyclophilin as a novel allergen from Platanus orientalis pollens by mass spectrometry. J Biosci Bioeng 2009;107:215-217

Oriental plane trees are an important source of airborne allergens in cities of southwest Asia. In spite of extensive studies on Platanus acerifolia allergy, there are no reports on the molecular characterization of pollen allergens from Platanus orientalis trees. In this study, a newly recognized member of cyclophilin family with a molecular weight of 18 kDa was identified as being partly responsible for IgE reactivity of P. orientalis pollen extract.

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[37] - Hunzelmann N, Baratli J, Asturias J. High prevalence of sensitization to plane allergens in Cologne. Allergy 2007;62(suppl. 83):171

Background: Platanus acerifolia pollen is an important contributor to allergic symptoms in South European and American cities, particularly in Southern Europe. To date published data only describe South-European cohorts, whereas little is known about the sensitization to plane pollen in Germany. Therefore we analyzed the frequency and pattern of sensitization to plane pollen in patients seen at the allergy outpatient clinic of the University Hospital of Cologne. Methods: 668 patients were tested by skin-prick tests (SPT). In addition, further analysis was carried out by determination of specific IgE-reactivity using the phadia ImmunoCAP system and reactivity to the major known recombinant allergens from P. Acerifolia i.e. Pla a 1, Pla a 2, plane profilin by EAST. In patients unreactive by EAST or CAP westernblot analysis was performed. Results: The prevalence of sensitization to P. acerifolia among 668 patients was 13,2% (88 patients). 7,5% (50) showed positive IgE-reactivity by SPT, 3,6% (24) by ImmunoCAP, 2,1% (14) to both. 75% of sensitized patients showed a total IgE > 100 kU/l. Furthermore, 35 patients positive by ImmunoCAP, were tested for specific IgE levels against recombinant Pla a 1, natural Pla a 2, and natural plane profilin. The test showed no clear concordance between ImmunoCAP and EAST, some of the sera had a positive CAP but no IgE-binding against purified allergens. Sera investigated by Western blotting against plane pollen extract demonstrated widespread IgE-reactivity and IgE-binding pattern. Conclusion: The results demonstrate a marked prevalence of plane sensitization in a central European city. Interestingly the complex IgE reactivity pattern found on sensitized patients parallels previous results described in southern Europe.

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[38] - Malandain H, Giroux F, Cano Y. The influence of carbohydrate structures present in common allergen sources on specific IgE results. Eur Ann Allergy Clin Immunol 2007;39:216-220

BACKGROUND: Cross-reactive carbohydrate determinants (CCD) are well known interferants in specific IgE assays (sIgE). Glyco-epitopes are not restricted to CCD and extracts used to prepare in vitro tests contain many other glycoproteins able to bind glycan-specific IgE. The overall amounts of IgE-bindable glycan structures in allergen sources are unknown . OBJECTIVE: We aimed at quantifying the influence of N-glycan structures on IgE reactivity to commonly tested allergen sources . METHODS: IgE reactivity to 51 allergen extracts, one purified natural allergen and 10 recombinant allergens was measured on Phadia UniCAP system using 2 sera demonstrating significant levels of glycan-related IgE reactivity. Immobilized bromelain and horseradish peroxidase (HRP) were used to capture N-glycan-specific IgE from these sera. Residual IgE reactivity was measured for 42 allergen sources and 4 recombinant/purified allergens . RESULTS: An obviously excessive number of positive CAP-results were obtained with both sera, especially for plant-based allergen sources. Capture of glycan-specific IgE led to a decrease of serum IgE ractivity, variable among allergen sources and between sera. Among others, peanut results were proven largely interfered by the presence of glycan-specific IgE. Unexpectedly some allergen sources showed a slight influence of glycan-related reactivity, such as cockroach, mosquito, mussel, shrimp and domestic mites . CONCLUSION: In patients sensitized to pollens or to Hymenoptera venoms sIgE results should be interpreted with caution. One cannot substract the result of a glyco-reporter test (bromelain and/or HRP) in order to compute glycan-free slgE results for common allergen sources like peanuts. As long as the demonstration of a significant role for glycan structures in clinical allergic reactions is lacking, a simple pre-treatment able to discard glycan-specific IgE from serum would be useful to improve accuracy of in vitro diagnostic tests.

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[39] - Tella R, Dall'Aglio M, San Miguel-Moncin M, Alonso R, Bartra J, Fernandez-Parra B, et al. Relationship between Platanus acerifolia pollinosis and food allergy in Barcelona Area. AAAAI 60th Annual Meeting, San Francisco, 19-23 March 2004, Poster n°480

Rationale Association of certain pollinosis with plant-derived food allergy has been demonstrated in several studies. The aim of this study was to analyze the clinical features and the prevalence of food allergy comparing patients sensitized to different pollen allergens (Platanus acerifolia versus other prevalent pollen allergies in our region). Methods From 6739 consecutive patients attending an allergy outpatient clinic during the last 10 years, 1477 suffered pollen allergies. Seventy-three (4.9%) patients suffered asthma, 919 rhinoconjunctivitis (62.2%) and 484 (32.8%) asthma and rhinoconjunctivitis. Patients were divided in a group sensitized to Platanus pollen (n=218, 14.7%), sensitized to grass (n=601, 40.6%) and sensitized to both pollens (n=122, 8.2%). We evaluated food allergy by clinical history, skin prick test +/ˆ challenge tests if proceeds in every group, and we compared which kind of food was implicated in each pollinosis. Results A 28.8% of Platanus sensitized patients suffered food allergy, while this fact only appeared in a 6.4% of the grass sensitized group (p<0.001). In the group of patients sensitized to both pollens, the prevalence of food allergy was similar to the Platanus sensitized group (27.8%). Regarding the clinical features of food allergy found in every group, systemic symptoms appeared in a 47.6% of Platanus group, a 38.9% of grass group and a 35.2% of patients from the group sensitized to both pollens. Conclusions Patients with pollinosis to Platanus could present more frequently food allergy compared with other pollinosis. Moreover, systemic reactions caused by food allergy are more frequent in patients sensitized to Platanus than in the other groups.

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[40] - Miralles JC, Caravaca F, Guillén F, Lombardero M, Negro JM. Cross-reactivity between Platanus pollen and vegetables. Allergy 2002;57:146-149

Background: Several associations have been described between tree and plant pollens and certain foods. The objective of this study is to verify whether there is cross-reactivity between Platanus pollen and vegetable origin foods. Methods: We selected 56 patients allergic to vegetable foods and subjected them to cutaneous tests with aeroallergens and vegetable foods. A statistical analysis was performed to evaluate the association of Platanus pollen with foods and with other aeroallergens. Later, a specific IgE determination was performed as well as a RAST (radioallergosorbent) inhibition experiment, to verify the existence of cross-reactivity in vitro. Results: In the cutaneous tests we found a positive correlation between Platanus pollen and hazelnut, peanut, banana and celery. The results of the RAST inhibition experiment indicate an important cross-reactivity between the pollen of Platanus acerifolia and hazelnut and banana fruit, and an intermediate cross-reactivity with celery and peanut. Conclusion: We have described an association between the pollen of the Platanus tree and some vegetable foods such as hazelnut, banana, peanut and celery. This association could be explained by the in vitro IgE cross-reactivity detected.

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[41] - Enrique E, Alonso R, Bartolome B, San Miguel-Moncin M, Bartra J, Fernandez-Parra B, et al. IgE reactivity to profilin in platanus acerifolia pollen-sensitized subjects with plant-derived food allergy. J Investig Allergol Clin Immunol 2004;14:335-342

BACKGROUND: The presence of profilin-specific IgE antibodies is a cause of cross-reactivity between botanically-unrelated allergen sources. Recently, the association between Platanus acerifolia pollinosis and plant-derived food allergy has been described. The aim of this study was to ascertain whether the P. acerifolia profilin is involved in such cross-reactivity. METHODS: Twenty-three patients suffering from Platanus acerifolia pollinosis and plant-derived food allergy were evaluated in an allergy department. Specific IgE levels to P. acerifolia pollen, P. acerifolia profilin and food extracts were measured. Molecular masses of IgE-binding proteins were calculated by Western blotting and cross-reactivity studies among P. acerifolia profilin and different food extracts were evaluated by Enzyme AllergoSorbent Test (EAST)-inhibition assays. Also, EAST-inhibition assays with the two known P. acerifolia allergens, Pla a 1 and Pla a 2, were performed. RESULTS: Surprisingly, a high IgE-binding prevalence (90%) of P. acerifolia profilin was found. EAST-inhibition showed high inhibition values when Platanus acerifolia pollen extract was used as free phase and plant-derived food extracts as solid phase, whereas the other way round showed low inhibition values. IgE reactivity to profilin was studied using a pool of patient sera, by EAST-inhibition assays with hazelnut, apple peel, peanut, chickpea and peanut extracts as solid phase and no inhibition was obtained when P. acerifolia profilin was used as inhibitor phase. The same results were obtained when purified Pla a 1 and Pla a 2 were also used as inhibitor phase. CONCLUSIONS: The clinical association observed between Platanus acerifolia pollen and plant-derived food could be explained by the in vitro IgE cross-reactivity detected by EAST-inhibition. However, it appears that neither P. acerifolia profilin nor the two major allergens described (Pla a 1 and Pla a 2) can explain such a strong cross-reactivity.

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[42] - Schocker F, Lüttkopf D, Scheurer S, Petersen A, Cisteró-Bahima A, Enrique E, et al. Recombinant lipid transfer protein Cor a 8 from hazelnut: A new tool for in vitro diagnosis of potentially severe hazelnut allergy. J Allergy Clin Immunol 2004;113:141-147

BACKGROUND: Cor a 1.04 has been identified as the major hazelnut allergen in 65 European patients with positive double-blind, placebo-controlled food challenge results to hazelnut. Recently, the 11S globulin Cor a 9 was shown to be a pollen-independent hazelnut allergen in the United States, whereas preliminary data suggest the lipid transfer protein (LTP) as an important birch pollen-unrelated hazelnut allergen in Europe . OBJECTIVE: We sought to recruit a group of European patients allergic to hazelnut without birch pollen allergy and to identify and clone the major food allergen(s) in this study population . METHODS: We recruited 26 such Spanish patients, including 10 patients with anaphylaxis. IgE immunoblotting was performed with hazelnut extract. Hazelnut LTP Cor a 8 was cloned by using a PCR strategy, purified, and subjected to IgE immunoblotting. Recombinant Cor a 8, rCor a 1.0401, and rCor a 2 (profilin) were further investigated by means of enzyme allergosorbent test. Immunoblot inhibition experiments were used to compare the immunologic properties of natural and recombinant LTP . RESULTS: A 9-kd major allergen was identified in hazelnut extract. Cloning, sequencing, heterologous expression, and inhibition experiments identified it as an LTP. The prevalence of specific IgE antibody reactivity to LTP was 62% in hazelnut extract and 77% when recombinant LTP was tested by means of immunoblotting. IgE immunoblot inhibition with hazelnut extract showed that natural Cor a 8 and rCor a 8 shared identical epitopes. Only one patient had positive reactivity to Cor a 1.04, and no patients had positive reactivity to Cor a 2. Two sera bound to high-molecular-weight allergens. The LTP was denominated as Cor a 8 and submitted to the allergen database of the World Health Organization/International Union of Immunological Societies Allergen Nomenclature Subcommittee . CONCLUSIONS: Cor a 8 is a relevant allergen for a majority of Spanish patients with hazelnut allergy that can cause severe allergic reactions.

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[43] - San Miguel-Moncin M, Krail M, Scheurer S, Enrique E, Alonso R, Conti A, et al. Lettuce anaphylaxis: identification of a lipid transfer protein as the major allergen. Allergy 2003;58:511-517

BACKGROUND: Allergy to plant-derived foods is associated with birch pollinosis in central and northern Europe. Symptoms elicited are usually limited to the oropharyngeal system. By contrast, in the Mediterranean area, allergy to the same foods manifests more frequently with systemic reactions caused by nonspecific lipid transfer proteins (nsLTP), independently of an associated pollinosis . OBJECTIVE: We sought to investigate the pattern of immunoglobulin E (IgE) binding protein bands implicated in lettuce allergy, in particular the presence of an nsLTP . METHODS: Consecutive lettuce allergic patients were selected. Determination of serum-specific IgE, immunoblot, and inhibition experiments were performed in order to study the pattern of IgE binding proteins and the potential cross-reactivity to pollens. Inhibition studies with recombinant allergens were conducted to identify the lettuce allergens. The major allergen was subjected to N-terminal amino acid sequencing . RESULTS: Fourteen patients were diagnosed as being allergic to lettuce. All were sensitized to Platanus pollen. Ten of them showed specific IgE to a lettuce protein of 9-kDa. The IgE binding to this protein was completely inhibited by the cherry-LTP and peach extract. The N-terminal sequence of the 9-kDa protein showed a high degree of amino acid sequence identity to other nsLTPs. A clear partial cross-reactivity was observed between lettuce-LTP and Platanus-pollen extract . CONCLUSIONS: An LTP has been demonstrated to be a major allergen in patients suffering from lettuce allergy.

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[44] - Miralles JC, Caravaca F, Guillén F, Lombardero M, Negro JM. Cross-reactivity between Platanus pollen and vegetables. Allergy 2002;57:146-149

Background: Several associations have been described between tree and plant pollens and certain foods. The objective of this study is to verify whether there is cross-reactivity between Platanus pollen and vegetable origin foods. Methods: We selected 56 patients allergic to vegetable foods and subjected them to cutaneous tests with aeroallergens and vegetable foods. A statistical analysis was performed to evaluate the association of Platanus pollen with foods and with other aeroallergens. Later, a specific IgE determination was performed as well as a RAST (radioallergosorbent) inhibition experiment, to verify the existence of cross-reactivity in vitro. Results: In the cutaneous tests we found a positive correlation between Platanus pollen and hazelnut, peanut, banana and celery. The results of the RAST inhibition experiment indicate an important cross-reactivity between the pollen of Platanus acerifolia and hazelnut and banana fruit, and an intermediate cross-reactivity with celery and peanut. Conclusion: We have described an association between the pollen of the Platanus tree and some vegetable foods such as hazelnut, banana, peanut and celery. This association could be explained by the in vitro IgE cross-reactivity detected.

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[45] - Enrique E, Cisteró-Bahíma A, Bartolomé B, Alonso R, San Miguel-Moncín MM, Bartra J, et al. Platanus acerifolia pollinosis and food allergy. Allergy 2002;57:351-356

Background:In Mediterranean areas, oral allergy syndrome (OAS) occurs independently of an associated birch pollinosis; moreover, on occasions it presents with no other associated pollinosis. The aim of this study was to assess the possible association of OAS with Platanus acerifolia pollinosis. Methods:We evaluated consecutive patients seen for pollinosis in an allergy department. Seven hundred and twenty patients were selected on the basis of seasonal or perennial rhinitis, or asthma, or both. Respiratory and food allergies were studied in all patients. Clinical history was recorded and examinations and skin prick tests were performed with a battery of available common inhalant allergens and plant-derived food allergens. Specific IgE levels to P.acerifolia pollen extract and food allergens tested were measured. Molecular masses of the IgE-binding proteins and cross-reactivity among the P. acerifolia pollen and different food extracts were also determined. Results:Of the 720 patients evaluated, 61 (8.48%) were sensitized to P.acerifolia pollen. Food allergy was observed in 32 (52.45%) of the 61 patients sensitized to P.acerifolia pollen. Food allergens most frequently implicated were hazelnuts, peach, apple, peanuts, maize, chickpea and lettuce. Enzyme allergosorbent (EAST)-inhibition showed high inhibition values when P.acerifolia pollen extract was used as free phase. On the contrary low inhibition was observed when plant-derived food allergens were used as free phase and P.acerifolia pollen extract as solid phase. Conclusions:Cross-reactivity was observed among P.acerifolia pollen and plant-derived foods. OAS in these patients may have been caused by primary respiratory sensitization. [= AAAAI 58th Annual Meeting, New York, 1-6 March, 2002, Poster n°936]

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[46] - Enrique E, Alonso R, Bartolome B, San Miguel-Moncin M, Bartra J, Fernandez-Parra B, et al. IgE reactivity to profilin in platanus acerifolia pollen-sensitized subjects with plant-derived food allergy. J Investig Allergol Clin Immunol 2004;14:335-342

BACKGROUND: The presence of profilin-specific IgE antibodies is a cause of cross-reactivity between botanically-unrelated allergen sources. Recently, the association between Platanus acerifolia pollinosis and plant-derived food allergy has been described. The aim of this study was to ascertain whether the P. acerifolia profilin is involved in such cross-reactivity. METHODS: Twenty-three patients suffering from Platanus acerifolia pollinosis and plant-derived food allergy were evaluated in an allergy department. Specific IgE levels to P. acerifolia pollen, P. acerifolia profilin and food extracts were measured. Molecular masses of IgE-binding proteins were calculated by Western blotting and cross-reactivity studies among P. acerifolia profilin and different food extracts were evaluated by Enzyme AllergoSorbent Test (EAST)-inhibition assays. Also, EAST-inhibition assays with the two known P. acerifolia allergens, Pla a 1 and Pla a 2, were performed. RESULTS: Surprisingly, a high IgE-binding prevalence (90%) of P. acerifolia profilin was found. EAST-inhibition showed high inhibition values when Platanus acerifolia pollen extract was used as free phase and plant-derived food extracts as solid phase, whereas the other way round showed low inhibition values. IgE reactivity to profilin was studied using a pool of patient sera, by EAST-inhibition assays with hazelnut, apple peel, peanut, chickpea and peanut extracts as solid phase and no inhibition was obtained when P. acerifolia profilin was used as inhibitor phase. The same results were obtained when purified Pla a 1 and Pla a 2 were also used as inhibitor phase. CONCLUSIONS: The clinical association observed between Platanus acerifolia pollen and plant-derived food could be explained by the in vitro IgE cross-reactivity detected by EAST-inhibition. However, it appears that neither P. acerifolia profilin nor the two major allergens described (Pla a 1 and Pla a 2) can explain such a strong cross-reactivity.

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[47] - Fernández-Rivas M, Bolhaar S, González-Mancebo E, Asero R, van Leeuwen A, Bohle B, et al. Apple allergy across Europe: How allergen sensitization profiles determine the clinical expression of allergies to plant foods. J Allergy Clin Immunol 2006;118:481-488

BACKGROUND: Allergy to a plant food can either result from direct sensitization to that food or from primary sensitization to pollen, latex, or another food . OBJECTIVE: We sought to investigate the primary sensitizers in apple allergy across Europe, the individual allergens involved, and whether these differences determine the clinical presentation . METHODS: Patients (n = 389) with positive case histories and skin prick test responses to fresh apple were selected in the Netherlands, Austria, Italy, and Spain. Skin prick tests and RASTs to a panel of pollens and plant foods were performed, as well as RASTs to Bet v 1 and the apple allergens Mal d 1, 2, 3, and 4 . RESULTS: In the Netherlands, Austria, and Italy apple allergy is mild (>90% isolated oral symptoms) and related to birch pollinosis and sensitization to Bet v 1 and its apple homologue, Mal d 1, which has an odds ratio of local reactions of 2.85 (95% CI, 1.47-5.55). In Spain apple allergy is severe (>35% systemic reactions) and related to peach allergy and sensitization to Mal d 3 (nonspecific lipid transfer protein), which has an odds ratio of systemic reactions of 7.76 (95% CI, 3.87-15.56) . CONCLUSION: The analysis of individual apple allergens in a clinical context has provided insight into the sensitization pathway and into the intrinsic risk an allergen bears to induce mild or severe food allergy. CLINICAL IMPLICATIONS: Information on the sensitization pathway is essential to develop preventive strategies in food allergy. The application of individual food allergens with a known intrinsic risk will improve the prognostic value of diagnostic tests.

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[48] - Figueroa J, Blanco C, Dumpiérrez AG, Almeida L, Ortega N, Castillo R, et al. Mustard allergy confirmed by double-blind placebo-controlled food challenges: clinical features and cross-reactivity with mugwort pollen and plant-derived foods. Allergy 2005;60:48-55

BACKGROUND: Mustard IgE-mediated allergy is supposed to be a rare cause of food allergy, and its clinical features and cross-reactivities have not been fully elucidated . METHODS: A prospective study was carried out, recruiting mustard allergic patients, and paired control subjects. A clinical questionnaire was administered, and skin-prick tests (SPT) with panels of aeroallergens and foods, serum extraction for in vitro tests and double-blind placebo-controlled food challenges (DBPCFC) were performed . RESULTS: Thirty-eight mainly adult patients, with 10.5% reporting systemic anaphylaxis, were included in the study [age (mean +/- SD): 21.9 +/- 8.6 years]. DBPCFC were performed in 24 patients, being positive in 14 cases (58.3%). Patients with positive outcome showed significantly greater mustard SPT than those with negative outcome (8.2 +/- 3.7 vs 5.3 +/- 2.4 mm, P <0.05), and the receiver-operating characteristic (ROC) curve analysis yielded a cut-off value for mustard commercial SPT of 8 mm, with a specificity of 90% (95% CI, 55.5-98.3), and a sensitivity of 50% (95% CI, 23.1-76.9). A significant association between mustard hypersensitivity and mugwort pollen sensitization was found (97.4% of patients), with partial cross-reactivity demonstrated by UniCAP System inhibition assays. All patients showed sensitization to other members of Brassicaceae family, and cross-reactivity among them was also confirmed. Moreover, significant associations with nut (97.4%), leguminous (94.7%), corn (78.9%), and Rosaceae fruit (89.5%) sensitizations were also shown. Around 40% of these food sensitizations were symptomatic, including food-dependent exercise-induced anaphylaxis in six patients . CONCLUSIONS: Mustard allergy is a not-uncommon disorder that can induce severe reactions. Significant associations with mugwort pollinosis and several plant-derived food allergies are demonstrated, suggesting a new mustard-mugwort allergy syndrome. A relationship between this syndrome and food-dependent exercise-induced anaphylaxis is also reported.

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[49] - Ballmer-Weber BK, Scheurer S, Fritsche P, Enrique E, Cistero-Bahima A, Haase T, et al. Component-resolved diagnosis with recombinant allergens in patients with cherry allergy. J Allergy Clin Immunol 2002;110:167-173

BACKGROUND: In pollen-related food allergy, extracts for skin prick tests (SPTs) are often not standardized, and the test reliability is affected by false-negative reactions . OBJECTIVE: We sought to evaluate a panel of recombinant allergens (RAs) derived from one allergenic food for use in component-resolved in vivo diagnosis, taking cherry as a model food . METHODS: Seventy-nine subjects were included in the study: 24 Swiss patients (group 1) with a positive double-blind placebo-controlled food challenge result to cherries, 23 patients with birch pollen allergy but without cherry allergy (group 2), 23 nonatopic subjects (group 3), and 9 Spanish patients with a history of a cherry allergy (group 4). SPTs were performed in duplicate by using recombinant cherry allergens (Bet v 1-related allergen: recombinant (r) Pru av 1; profilin: rPru av 4; and lipid transfer protein: rPru av 3) in concentrations of 10, 50, and 100 microg/mL. Furthermore, IgE reactivity to rPru av 1, rPru av 4, and rPru av 3 was assessed by means of immunoblot analysis . RESULTS: SPT responses with rPru av 1, rPru av 4, and rPru av 3 were positive in 92%, 17%, and 4% of the patients in group 1; in 74%, 30%, and 0% of the patients in group 2; in 0%, 22%, and 89% of the patients in group 4; and negative for all nonatopic subjects (group 3). Thus the sensitivity of a positive SPT response to at least one of the 3 RAs was 96%. The specificities, negative predictive values, and positive predictive values with the 3 RAs were 100%, 96%, and 100% if calculated in relation to the nonatopic control group but 17%, 79%, and 60% when calculated in relation to the control group with birch pollen allergy. The correlation between SPT and immunoblotting results was excellent. Sensitization to rPru av 3 was associated with more severe symptoms than sensitization to rPru av 1 . CONCLUSIONS: SPTs with RAs proved to be highly sensitive for diagnosis of cherry allergy. Component-resolved in vivo diagnosis with standardized amounts of stable RAs allows us to determine sensitization patterns directly, to correlate them with severity of clinical symptoms, and to analyze geographic differences.

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[50] - Lauer I, San Miguel-Moncin M, Abel T, Foetisch K, Hartz C, Fortunato D, et al. Identification of a plane pollen lipid transfer protein (Pla a 3) and its immunological relation to the peach lipid-transfer protein, Pru p 3. Clin Exp Allergy 2007;37:261-269

Background An association between plane tree pollen allergy and plant food allergy has been described, but the cross-reacting allergens have not yet been identified. The aim of this study was the identification of homologous non-specific lipid-transfer proteins (nsLTPs) in plane pollen, and to investigate its immunological relationship with the peach LTP, Pru p 3. Methods Three different patient groups were recruited in Spain: 22 plane pollen-allergic patients without food allergy (A), 36 plane pollen-allergic patients with peach allergy (B) and 10 peach-allergic patients without plane pollen allergy (C). Proteins from plane pollen extract were fractionated by ion-exchange and reversed-phase chromatography. Further methods applied were N-terminal amino acid sequence analysis, immunoblotting, enzyme allergosorbent test, CAP and basophil histamine release assays. Results A 10 kDa IgE-reactive protein was purified from plane pollen and identified as nsLTP. Pla a 3 was characterized as a minor allergen (27.3%) in plane pollen-allergic patients without food allergy (A) and as a major allergen in plane pollen-allergic patients with peach allergy (B) showing a prevalence of IgE-reactivity of 63.8%. Group B contained patients sensitized to Pru p 3 without IgE-reactivity to plane-LTP (16.6%). By contrast, Pla a 3 IgE-reactive patients without sensitization to Pru p 3 could be found (16.6%). The sera of patients sensitized to both LTPs (50%), Pla a 3 and Pru p 3, showed different biological activity in histamine release assay: depending on individual patient's sera tested, Pla a 3 showed a similar, a stronger or a weaker allergenic potency in comparison with Pru p 3. Conclusions Plane LTP is a major allergen in plane pollen-allergic patients with peach allergy recruited in the Mediterranean area. The results of histamine release tests and different IgE-binding profiles pointed towards the existence of species-specific IgE epitopes. Likewise, no general conclusion on the sensitizer could be made.

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[51] - Lauer I, San Miguel-Moncín M, Hartz C, Cistero-Bahima A, Conti A, Vieths S, et al. The lipid transfer protein (LTP) from Platanus acerifolia pollen, Pla a 3, is a major allergen and cross-reacts with food LTPs. EAACI 23th Congress, Amsterdam, 12-16 June, 2004, Poster n°1001

Background: Plane trees such as Platanus acerifolia are an important source of airborne allergens in many cities of Europe. Until now, only two major allergens of plane pollen, Pla a 1 and Pla a 2, have been characterised. In the Mediterranean area food-allergic patients are frequently sensitised to non-specific LTPs (nsLTPs). Sensitisation to food LTPs is not related to birch pollen allergy, and patients suffer from more severe reactions, in addition to OAS. An association between plane tree pollen allergy and plant food allergy was described, but the cross-reacting allergens have not been identified. The aim of this study was identification and characterisation of the nsLTP from plane pollen, and to investigate its immunological relationship to food LTPs from lettuce, cherry and hazelnut. Methods: Plane pollen extract was fractionated by ion exchange chromatography. Plane pollen LTP was identified by N-terminal amino acid sequence analysis. The prevalence of IgE reactivity to plane LTP was determined by immunoblotting using 45 Spanish plane pollen allergic patients with various plant food allergies and 14 plane pollen sensitised patients from Germany. In addition, sensitisation to LTPs from nuts, fruits and vegetables and their cross-reactivity to plane LTP was evaluated by immunoblotting and IgE inhibition experiments. Results: A 10 kDa IgE-reactive protein from plane pollen was partially purified and identified by N-terminal amino acid sequencing as nsLTP (Pla a 3). The N-terminal sequence of Pla a 3 showed the highest aa-ID of 58% with that of Lac s 1 (lettuce LTP). Pla a 3 was cross-reactive with food LTPs. Pla a 3 showed a prevalence of IgE reactivity of 66.6% (30/45) in Spanish plane pollen allergic patients. In contrast, Pla a 3 sensitisation was not relevant in plane pollen sensitised patients from Germany. We observed only partial correlation between sensitisation to plane pollen LTP and LTPs from lettuce, cherry and hazelnut. On the one hand, there were Spanish patients sensitised to food LTPs without plane pollen allergy, on the other hand, food LTP allergic patients without sensitisation to Pla a 3 were found. Conclusions: We have characterised the plane LTP as major allergen in plane pollen allergic patients with concomitant allergy to plant-derived foods recruited in the Mediterranean area. In Spanish patients sensitisation to food LTPs is not associated exclusively to the plane pollen LTP.

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[52] - Skypala I, Calderon M, Leeds A, Durham S. Pollen-food syndrome in United Kingdom subjects - which aeroallergens are commonly involved ? Allergy 2007;62(suppl. 83):358

Pollen food syndrome (PFS) is common in individuals with birch pollen sensitivity who present with hay fever symptoms in the spring. The aim of this study was to evaluate which pollen(s), apart from birch, are associated with PFS in UK-based subjects, and whether aero-allergen sensitivities differ in those subjects with springtime hay fever with/without associated PFS. Adult subjects reporting hay fever symptoms between March and May were included in the study. All subjects underwent skin prick testing with tree (birch, oak, beech, plane and three trees ˜ birch, alder and hazel), timothy grass, mugwort, latex, cat and house dust mite (D. pteronyssinus) allergens (ALK Abelló Denmark). A diagnosis of PFS was established for each subject using a combination of results from a standardised medical assessment, prick by prick testing with suspect foods and oral food challenge. 119 subjects completed the study; 92% were sensitised to one or more aeroallergens. Although all subjects reported spring and/or summer time hay fever, only 85% of the cohort were sensitised to pollen. Grass was the commonest sensitising pollen allergen (68%), followed by birch (64%), oak and three trees (53%), beech (34%), plane (26%) and mugwort (18%). Approximately half of the cohort were sensitised to cat and/or house dust mite, but less than 10% to latex. Subjects diagnosed with PFS had a significantly greater number of positive tests for birch, oak, beech (p<0.001) and grass (p<0.05) than those without PFS, but there was no significant difference between the groups for the other aeroallergens. Polysensitisation to pollen allergens occurred in 93% of the group diagnosed with PFS compared to 37% of the group without PFS. No aeroallergen sensitivities were individually or collectively significant predictors of the diagnosis of PFS although a combination of birch, grass and oak gave the best standardised beta coefficient. UK subjects with springtime hay fever are most likely to be sensitised to birch, grass and oak. The frequency of sensitivities to these allergens is greater in those who additionally have PFS. In contrast, subjects with/without PFS do not differ in their rate of sensitisation to cat and house dust mite. Sensitisation to pollen cannot predict who with springtime hay fever will have PFS, but those with PFS are more likely to be co-sensitised to several tree and grass pollens.

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[53] - Enrique E, Cisteró-Bahíma A, Bartolomé B, Alonso R, San Miguel-Moncín MM, Bartra J, et al. Platanus acerifolia pollinosis and food allergy. Allergy 2002;57:351-356

Background:In Mediterranean areas, oral allergy syndrome (OAS) occurs independently of an associated birch pollinosis; moreover, on occasions it presents with no other associated pollinosis. The aim of this study was to assess the possible association of OAS with Platanus acerifolia pollinosis. Methods:We evaluated consecutive patients seen for pollinosis in an allergy department. Seven hundred and twenty patients were selected on the basis of seasonal or perennial rhinitis, or asthma, or both. Respiratory and food allergies were studied in all patients. Clinical history was recorded and examinations and skin prick tests were performed with a battery of available common inhalant allergens and plant-derived food allergens. Specific IgE levels to P.acerifolia pollen extract and food allergens tested were measured. Molecular masses of the IgE-binding proteins and cross-reactivity among the P. acerifolia pollen and different food extracts were also determined. Results:Of the 720 patients evaluated, 61 (8.48%) were sensitized to P.acerifolia pollen. Food allergy was observed in 32 (52.45%) of the 61 patients sensitized to P.acerifolia pollen. Food allergens most frequently implicated were hazelnuts, peach, apple, peanuts, maize, chickpea and lettuce. Enzyme allergosorbent (EAST)-inhibition showed high inhibition values when P.acerifolia pollen extract was used as free phase. On the contrary low inhibition was observed when plant-derived food allergens were used as free phase and P.acerifolia pollen extract as solid phase. Conclusions:Cross-reactivity was observed among P.acerifolia pollen and plant-derived foods. OAS in these patients may have been caused by primary respiratory sensitization. [= AAAAI 58th Annual Meeting, New York, 1-6 March, 2002, Poster n°936]

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[54] - Crespo JF, Retzek M, Fötisch K, Sierra-Maestro E, Cid-Sanchez AB, Pascual CY, et al. Germin-like protein Cit s 1 and profilin Cit s 2 are major allergens in orange (Citrus sinensis) fruits. Mol Nutr Food Res 2006;50:282-290

Oranges are clinically relevant allergenic foods. To date, orange allergens have not been characterized in detail. The study is aimed at analyzing the sensitization profile in orange-sensitized subjects with and without clinical allergy, and to identify orange allergens. Fifty-six sensitized subjects with self-reported reactions to orange were grouped into reactors (anaphylaxis or multiple episodes of immediate reactions and/or positive challenge tests) and non-reactors (negative open food challenge tests). Allergens were characterized by IgE immunoblotting, N-terminal sequencing, IgE-inhibition assays, and mediator release assays were performed to determine the allergenic potency of orange profilin. Of 56 subjects, 23 were classified as orange allergic showing mainly an oral allergy syndrome. Of 23 subjects classified as orange allergic, 22 were sensitized to profilin, Cit s 2. In patients with mono-sensitization to profilin in vitro histamine releases up to 75% from basophils were induced using orange extract and purified plant profilins. Of the allergic patients 78% were sensitized to germin-like protein, Cit s 1. Both allergens showed retained IgE reactivity in heat-processed orange juice. Interestingly, subjects with and without clinical allergy showed a comparable sensitization profile. Profilin and germin-like proteins are major orange allergens. The potential clinical relevance of orange profilin was indicated by its strong capacity to release histamine from basophils. However, a predominant sensitization to both allergens in subjects without symptoms also indicates a high frequency of clinically insignificant sensitization.

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[55] - Lauer I, San Miguel-Moncin M, Abel T, Foetisch K, Hartz C, Fortunato D, et al. Identification of a plane pollen lipid transfer protein (Pla a 3) and its immunological relation to the peach lipid-transfer protein, Pru p 3. Clin Exp Allergy 2007;37:261-269

Background An association between plane tree pollen allergy and plant food allergy has been described, but the cross-reacting allergens have not yet been identified. The aim of this study was the identification of homologous non-specific lipid-transfer proteins (nsLTPs) in plane pollen, and to investigate its immunological relationship with the peach LTP, Pru p 3. Methods Three different patient groups were recruited in Spain: 22 plane pollen-allergic patients without food allergy (A), 36 plane pollen-allergic patients with peach allergy (B) and 10 peach-allergic patients without plane pollen allergy (C). Proteins from plane pollen extract were fractionated by ion-exchange and reversed-phase chromatography. Further methods applied were N-terminal amino acid sequence analysis, immunoblotting, enzyme allergosorbent test, CAP and basophil histamine release assays. Results A 10 kDa IgE-reactive protein was purified from plane pollen and identified as nsLTP. Pla a 3 was characterized as a minor allergen (27.3%) in plane pollen-allergic patients without food allergy (A) and as a major allergen in plane pollen-allergic patients with peach allergy (B) showing a prevalence of IgE-reactivity of 63.8%. Group B contained patients sensitized to Pru p 3 without IgE-reactivity to plane-LTP (16.6%). By contrast, Pla a 3 IgE-reactive patients without sensitization to Pru p 3 could be found (16.6%). The sera of patients sensitized to both LTPs (50%), Pla a 3 and Pru p 3, showed different biological activity in histamine release assay: depending on individual patient's sera tested, Pla a 3 showed a similar, a stronger or a weaker allergenic potency in comparison with Pru p 3. Conclusions Plane LTP is a major allergen in plane pollen-allergic patients with peach allergy recruited in the Mediterranean area. The results of histamine release tests and different IgE-binding profiles pointed towards the existence of species-specific IgE epitopes. Likewise, no general conclusion on the sensitizer could be made.

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Les allergènes de platane

Diagnostic d’une pollinose au platane

Platane et allergie alimentaire

Rubriques

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