Les Pariétaires

[1] - Mari A, Artale A, Bonura A, Scala E, Colombo P. Par j 2 as a parietaria sensitisation marker allergen: definition of its use in epidemiology and diagnosis. Allergy Clin Immunol Int 2005;17(Suppl. 1):205

Several allergens have been identifed, characterised and cloned from Parietaria pollen. Par j 1 and Par j 2 represent allergenic molecules recognised by patients' IgE who are usually not sensitised to other pollen sources. The recombinant Par j 2 (rPar j 2) is now available for in vitro testing (Pharmacia, Sweden). We examined a cohort of subject with pollinosis symptoms. Subjects were from urban and non-urban areas, with different exposure levels to Parietaria. Patients underwent skin test (SPT) with a previously defined panel of pollen allergenic extracts and serum samples were obtained. IgE reactivity to Parietaria judaica extract (PjE) was compared to rPar j 2 reactivity. A cohort of 2993 pollinosis subject were identified, 37.1 % had a positive SPT to a PjE. IgE to PjE were detected in 96.3 % of 709 blood samples. rPar j 2 IgE positive samples were 82.2% of 573 PjE IgE positive samples. Differences were recorded when different patient subsets were considered. 92.5 % of Parietaria mono-sensitised subjects (n= 80); SPT and IgE positive had IgE to rParj 2. Similar results were recorded when Parietaria was associated to other pollens (92%; n=289), whereas only 64.2% were positive in the group of multiple pollen sensitised subjects (MPS; n=204). Differences were recorded in geographical distribution of rPar j 2 reactive subjects. A relatively small area with statistically significant lower prevalence rPar j 2 IgE positive subjects was identified when native subjects were considered. Correlating IgE results, PjE vs rPar j 2, the best result was achieved in the mono-sensitised and oligo-sensitised cohort again if compared to the MPS patient subset. When results from mono-sensitised subjects were plotted, three further subgroups were identified. A first group with almost overlapping values for both PjE and rPar j 2, a second with PjE values greater than rPar j 2 values, and a third group with the opposite finding. Representative sera from these three subgroups have been tested for both rPar j 1 and 2. All reacted with both. rPar j 2 can be assumed as a specific marker for exposure and sensitisation to Parietaria pollen. rPar j 2 evaluation allows both a more precise geographical mapping and for identification of Parietaria sensitised subjects within MPS subset. Further studies are needed to define whether a single molecule (rPar j 2) is suitable for both diagnosis and future molecule-based immunotherapy.

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[2] - Asturias JA, Gómez-Bayón N, Eseverri JL, Martínez A. IgE epitope analysis of Par j 1 and Par j 2, the major allergens of Parietaria judaica. EAACI 21th Congress, Naples, 1-5 June, 2002, Poster n°922

Parietaria judaica pollen is the main cause of allergy in the Mediterranean area with a prevalence of 60-80% in Italy and Greece and 25-40% in Spain and Southern France. This pollen contains at least nine allergens, and two of them, named Par j 1 and Par j 2, have been cloned and sequenced. Par j 1 is a 14.7 kDa protein with a prevalence of 95% among P. judaica allergic patients. Par j 2 is a protein of 11.3 kDa which reacted with IgE of 82% of the sera of P. judaica allergic subjects. Both proteins belong to the family of protein refereed to as non-specific lipid transfer proteins (ns-LTP). In the present study, we used overlapping synthetic peptides to identify major IgE-binding epitopes of Par j 1 and Par j 2 in patients with Parietaria pollen allergy. Serum samples were obtained from 17 patients with P. judaica allergy. All patients were chosen based on their clinical history, skin prick test reactivity, specific IgE class > 2, and immunoblot reactivity to 10-14 kDa Par j 1/2. Individual peptides were synthesized on a derivatized cellulose membrane using Fmoc chemistry (SPOTs). Homology modeling of the three-dimensional structure of Par j 1 and Par j 2 was generated using the Swiss-Model server. Six individual IgE-binding epitopes, of 6-10 residues long, were identified in Par j 1 corresponding to regions: Q19-P25, C30-G38, T42-A49, P72-D79, L82-C91, and P118-E125, and named epitopes 1-I to 1-VI, respectively. Eight overlapping IgE-binding epitopes was detected in Par j 2 corresponding to regions: C14-E21, V18-K27, C30-E39, E38-Q45, K46-R55, C52-I60, K73-T80, and S91-F98 and named epitopes 2-I to 2-VIII, respectively. There were no obvious amino acid sequence motifs shared by all the IgE-binding epitopes. The lineal IgE epitopes described were represented on the 3-D structure of both allergens based on the maize ns-LTP. Reactive regions fitted with the second and third a-helix and turns 1, 2 and 4. Amino acid sequence comparisons of these IgE reactive regions showed three epitopes with high homology in both allergens. The availability of three-dimensional model in combination with the known sequences of the IgE-binding sites allows the design of monovalent allergen fragments (haptens) which would not cross-link effector-cell-bound IgE and, therefore, could be used for local blocking therapy as well as for active immunization.

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[3] - Bonura A, Artale A, Marino M, Amoroso S, Marcucci F, Geraci D, et al. Cross-reactivity between Parietaria species using the major rPar j 1 and rPar j 2 allergens. Allergy Asthma Proc 2006;27:378-382

Parietaria pollen is one of the most important outdoor allergenic sources in all the Mediterranean countries, with a large number of subjects showing a positive skin prick test to both P. judaica and P. officinalis pollen extracts. A cross-reactivity between the two species has been already reported although few data are known at the molecular level. Twenty-five consecutive patients with Parietaria pollen allergy were selected on the basis of their clinical history. Skin prick test to P. judaica and officinalis extracts was performed. In vitro IgE measurement to both allergenic sources was performed by using a quantitative assay. ELISA inhibition experiments were made by using the rParj1 and rParj2 allergens. All the patients showed a positive skin prick test to both Parietaria species. Quantitative IgE measurement showed similar antibody concentration for P. judaica and P. officinalis extracts. ELISA inhibition experiments demonstrated that the cross-reactivity between the two species was due to the presence of the Parj1 and Parj2 allergens in the extracts with a high conserved IgE epitopes content. We conclude that P. judaica and P. officinalis pollens contain highly cross-reactive species-specific major allergens useful for the diagnosis and therapy of both allergenic sources.

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[4] - Amoresano A, Pucci P, Duro G, Colombo P, Costa MA, Izzo V, et al. Assignment of disulphide bridges in Par j 2.0101, a major allergen of Parietaria judaica pollen. Biol Chem 2003;384:1165-1172

Par j 2.0101, a major allergen of the Parietaria judaica pollen, was expressed in E. coli, purified to homogeneity and fully characterised both at the structural and the functional level. The recombinant rPar j 2.0101 protein showed an allergenic activity in histamine release, skin prick tests and capacity to bind IgE, almost identical to that of the native allergens purified from aqueous pollen extract. The complete pattern of S-S bridges of rPar j 2.0101 was determined by enzymatic digestion with endoproteinase Lys-C followed by mass spectrometric analysis of the resulting peptide mixtures. The eight cysteines occurring in the allergenic protein were found to be paired into the following four disulphides: Cys35-Cys83, Cys45-Cys60, Cys61-Cys106 and Cys81-Cys121. This structural information probes Par j 2.0101 to attain a 3-D fold consistent with that of the non-specific lipid transfer protein (ns-LTP) family and it represents an effective molecular basis to develop modified antigens by selective site-directed mutagenesis for immunotherapy.

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[5] - Bousquet J, Hewitt B, Guerin B, Dhivert H, Michel FB. Allergy in the Mediterranean area. II: Cross-allergenicity among Urticaceae pollens (Parietaria and Urtica). Clin Allergy 1986;16:57-64

Pollens of related species usually share common antigens and are cross-reacting but this is not always the case. The cross-allergenicity of two species of Urticaceae pollens (Parietaria and Urtica, nettle) was sought by means of RAST-inhibition and isoelectric focusing. A preliminary experiment in forty-two patients investigated by means of skin tests and RAST showed that most patients were either allergic to Parietaria or nettle and that both reactivities were uncommonly associated. The results of RAST-inhibition confirmed the total absence of cross-reactivity between these two related species. IEF patterns indicated that most proteins are in the acidic range of pH and did not reveal large discrepancies. Parietaria and Urtica pollens, although belonging to the same family and having close microscopic patterns, have very different allergenic activities. This study demonstrates that cross-allergenicity cannot be predicted.

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[6] - 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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[7] - Hemery ML, Verdier R, Daban P, Sellier N, Dujols P, Demoly P. Sensibilisation aux gants en latex poudrés: une prévalence élevée chez le personnel hospitalier. Presse Med 2005;34:1363-1369

INTRODUCTION: The prevalence of latex allergies in industrial countries has skyrocketed since the 1980s. Between 2.6 and 22% of hospital workers are diagnosed with latex allergy, which has been recognized as an occupational hazard in France since 1997. AIM: To assess the prevalence of latex allergy among Montpellier University Hospital Center staff . METHODS: From 1999 to 2002, we interviewed and conducted skin-prick tests on 537 hospital staff members from different departments and with different degrees of exposure to airborne latex allergens . RESULTS: Symptoms while using natural-rubber latex gloves (most often contact dermatitis) were reported by 88 (16.4%) staff members. Overall work-related allergic symptoms included rhinitis, reported by 65 (12.1%), contact urticaria by 28 (5.2%), and bronchial symptoms by 16 (1.1%). In all, sensitization to latex was identified in 7.1% of our staff, and this percentage was higher (11.3%) in units where latex gloves were used more often. Sensitization was associated with high latex exposure and atopy . CONCLUSION: This high rate of latex sensitization and the risk factors are similar to those already published. Based on this study, we have eliminated powdered latex gloves, as have many other hospitals.

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[8] - Belver MT, Caballero MT, Contreras J, Cabañas R, Sierra E, Madero R et al. Associations Among Pollen Sensitizations From Different Botanical Species in Patients Living in the Northern Area of Madrid. J Investig Allergol Clin Immunol 2007;17:157-159

OBJECTIVE: To determinate the existence of associations among sensitizations to antigens produced by pollen grains of different botanical species as assessed by skin prick tests in patients with respiratory disorders . METHODS: Six hundred twenty nine consecutive patients living in the northern area of Madrid who underwent clinical evaluation because of rhinoconjunctivitis, and/or asthma were studied. All patients were tested with a skin prick test using a battery of inhalants including pollens, dust mites, molds and danders. The exploratory multivariate technique of Multiple Correspondence Analysis was used to compare the homogeneity of sensitizations between groups. Of the 629 patients, 459 (73.0%) had positive skin prick tests to pollen and were selected as the study group . RESULTS: The most prevalent pollen sensitization was to Gramineae pollen (83.7%) followed by Oleaceae sensitisation (75.8%). Multiple Correspondence Analysis revealed the existence of an association among pollen sensitizations, showing that they clustered two groups: sensitizations to Gramineae, Oleaceae, Cupressaceae, Chenopodiaceae, Plantaginaceae (group I), and sensitizations to Betulaceae, Platanaceae, Compositae (group II). Sensitization to Parietaria was not included in any of the sensitization groups and showed an independent behaviour . CONCLUSION: Pollen sensitizations in our area cluster into two association groups which have not previously been reported.

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[9] - Mari A, Artale A, Bonura A, Scala E, Colombo P. Par j 2 as a parietaria sensitisation marker allergen: definition of its use in epidemiology and diagnosis. Allergy Clin Immunol Int 2005;17(Suppl. 1):205

Several allergens have been identifed, characterised and cloned from Parietaria pollen. Par j 1 and Par j 2 represent allergenic molecules recognised by patients' IgE who are usually not sensitised to other pollen sources. The recombinant Par j 2 (rPar j 2) is now available for in vitro testing (Pharmacia, Sweden). We examined a cohort of subject with pollinosis symptoms. Subjects were from urban and non-urban areas, with different exposure levels to Parietaria. Patients underwent skin test (SPT) with a previously defined panel of pollen allergenic extracts and serum samples were obtained. IgE reactivity to Parietaria judaica extract (PjE) was compared to rPar j 2 reactivity. A cohort of 2993 pollinosis subject were identified, 37.1 % had a positive SPT to a PjE. IgE to PjE were detected in 96.3 % of 709 blood samples. rPar j 2 IgE positive samples were 82.2% of 573 PjE IgE positive samples. Differences were recorded when different patient subsets were considered. 92.5 % of Parietaria mono-sensitised subjects (n= 80); SPT and IgE positive had IgE to rParj 2. Similar results were recorded when Parietaria was associated to other pollens (92%; n=289), whereas only 64.2% were positive in the group of multiple pollen sensitised subjects (MPS; n=204). Differences were recorded in geographical distribution of rPar j 2 reactive subjects. A relatively small area with statistically significant lower prevalence rPar j 2 IgE positive subjects was identified when native subjects were considered. Correlating IgE results, PjE vs rPar j 2, the best result was achieved in the mono-sensitised and oligo-sensitised cohort again if compared to the MPS patient subset. When results from mono-sensitised subjects were plotted, three further subgroups were identified. A first group with almost overlapping values for both PjE and rPar j 2, a second with PjE values greater than rPar j 2 values, and a third group with the opposite finding. Representative sera from these three subgroups have been tested for both rPar j 1 and 2. All reacted with both. rPar j 2 can be assumed as a specific marker for exposure and sensitisation to Parietaria pollen. rPar j 2 evaluation allows both a more precise geographical mapping and for identification of Parietaria sensitised subjects within MPS subset. Further studies are needed to define whether a single molecule (rPar j 2) is suitable for both diagnosis and future molecule-based immunotherapy.

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[10] - Gadermaier G, Dedic A, Obermeyer G, Frank S, Himly M, Ferreira F. Biology of weed pollen allergens. Curr Allergy Asthma Rep 2004;4:391-400

Weeds represent a heterogeneous group of plants, usually defined by no commercial or aesthetic value. Important allergenic weeds belong to the plant families Asteraceae, Amaranthaceae, Urticaceae, Euphorbiaceae, and Plantaginaceae. Major allergens from ragweed, mugwort, feverfew, pellitory, goosefoot, Russian thistle, plantain, and Mercurialis pollen have been characterized to varying degrees. Four major families of proteins seem to be the major cause of allergic reactions to weed pollen: the ragweed Amb a 1 family of pectate lyases; the defensin-like Art v 1 family from mugwort, feverfew, and probably also from sunflower; the Ole e 1-like allergens Pla l 1 from plantain and Che a 1 from goosefoot; and the nonspecific lipid transfer proteins Par j 1 and Par j 2 from pellitory. As described for other pollens, weed pollen also contains the panallergens profilin and calcium-binding proteins, which are responsible for extensive cross-reactivity among pollen-sensitized patients.

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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] - 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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[13] - Figueredo E, Cuesta-Herranz J, De-Miguel J, Lazaro M, Sastre J, Quirce S, et al. Clinical characteristics of melon (Cucumis melo) allergy. Ann Allergy Asthma Immunol 2003;91:303-308

BACKGROUND: Although melon is a frequent allergy-eliciting fruit, allergic reactions to melon have rarely been reported. OBJECTIVE: To evaluate and describe the clinical characteristics of melon allergy in melon-allergic patients. MATERIALS AND METHODS: We evaluated patients allergic to melon and a control group of patients allergic to pollen. The diagnosis of melon allergy was based on a convincing clinical history, positive skin test results (prick-by-prick test), and positive results on oral challenge tests to melon. RESULTS: A total of 161 patients were included in the study: 66 in the melon allergy group and 95 in the pollen control group. The melon allergy group included 35 female and 31 male patients with a mean age of 26.6 +/- 2.7 years (range, 5-61 years). Although all patients had oral symptoms, 13 (19.7%) of the patients had extraoral symptoms and none experienced generalized urticaria or anaphylaxis. Excluding other Cucurbitaceae fruits, peach, fig, and kiwi most frequently elicited positive skin test results and symptoms. Up to 23% of melon-allergic patients had a concomitant latex sensitization. Melon allergy was especially linked to pollen allergy, since all the melon-allergic patients were also allergic to pollen. Some differential features with respect to the pollen allergy control group were a higher prevalence of asthma (odds ratio [OR], 2.13; P < 0.05) and a statistical increase in the frequency of sensitization to several tree and weed pollens, including Ulmus (OR, 42.8) and Ambrosia (OR, 22.4). CONCLUSION: The most important conditions linked to melon allergy are pollen allergy (100%), allergy to other nonrelated fruits, mainly peach (up to 62%), and latex sensitivity (up to 23%). Some differential features of the pollinosis in melon allergy were a higher prevalence of asthma and a higher frequency of sensitization to several weed and tree pollens.

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[14] - Mari A, Artale A, Bonura A, Scala E, Colombo P. Par j 2 as a parietaria sensitisation marker allergen: definition of its use in epidemiology and diagnosis. Allergy Clin Immunol Int 2005;17(Suppl. 1):205

Several allergens have been identifed, characterised and cloned from Parietaria pollen. Par j 1 and Par j 2 represent allergenic molecules recognised by patients' IgE who are usually not sensitised to other pollen sources. The recombinant Par j 2 (rPar j 2) is now available for in vitro testing (Pharmacia, Sweden). We examined a cohort of subject with pollinosis symptoms. Subjects were from urban and non-urban areas, with different exposure levels to Parietaria. Patients underwent skin test (SPT) with a previously defined panel of pollen allergenic extracts and serum samples were obtained. IgE reactivity to Parietaria judaica extract (PjE) was compared to rPar j 2 reactivity. A cohort of 2993 pollinosis subject were identified, 37.1 % had a positive SPT to a PjE. IgE to PjE were detected in 96.3 % of 709 blood samples. rPar j 2 IgE positive samples were 82.2% of 573 PjE IgE positive samples. Differences were recorded when different patient subsets were considered. 92.5 % of Parietaria mono-sensitised subjects (n= 80); SPT and IgE positive had IgE to rParj 2. Similar results were recorded when Parietaria was associated to other pollens (92%; n=289), whereas only 64.2% were positive in the group of multiple pollen sensitised subjects (MPS; n=204). Differences were recorded in geographical distribution of rPar j 2 reactive subjects. A relatively small area with statistically significant lower prevalence rPar j 2 IgE positive subjects was identified when native subjects were considered. Correlating IgE results, PjE vs rPar j 2, the best result was achieved in the mono-sensitised and oligo-sensitised cohort again if compared to the MPS patient subset. When results from mono-sensitised subjects were plotted, three further subgroups were identified. A first group with almost overlapping values for both PjE and rPar j 2, a second with PjE values greater than rPar j 2 values, and a third group with the opposite finding. Representative sera from these three subgroups have been tested for both rPar j 1 and 2. All reacted with both. rPar j 2 can be assumed as a specific marker for exposure and sensitisation to Parietaria pollen. rPar j 2 evaluation allows both a more precise geographical mapping and for identification of Parietaria sensitised subjects within MPS subset. Further studies are needed to define whether a single molecule (rPar j 2) is suitable for both diagnosis and future molecule-based immunotherapy.

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

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[16] - Holgate ST, Jackson L, Watson HK, Ganderton MA. Sensitivity to Parietaria pollen in the Southampton area as determined by skin-prick and RAST tests. Clin Allergy 1988;18:549-556

The pollen of Parietaria species is a well-recognized and important inhalant allergen in the Mediterranean area. Parietaria judaica (Pellitory-of-the-Wall) is native to the U.K., flowering from June to September, but is not usually considered to be of any clinical importance by U.K. allergists. We skin tested 62 patients with a clinical history of summer seasonal respiratory symptoms and a control group of 11 patients with perennial respiratory symptoms only. Each was skin tested in duplicate with extracts of grass pollen, birch pollen, Parietaria pollen, Dermatophagoides pteronyssinus, Cladosporium, Alternaria, nettle pollen and negative and positive controls, and serum samples were collected for RAST assays for Parietaria and nettle. Eight of the 62 patients in the main group showed skin reactivity to Parietaria. Five of these eight had never visited the Mediterranean area and therefore it is possible that sensitization occurred in the U.K. Thirteen of the 62 patients were skin reactive to nettle but there was no correlation between skin reactivity to Parietaria and nettle. This supports a recent report that, despite their close botanical relationship, no antigenic cross-reactivity exists between the two species. No correlation was seen between skin reactivity and serum RAST activity to Parietaria or nettle. It is not known whether exposure to Parietaria pollen contributes to the seasonal symptoms of the patients found to be skin reactive. None of the 11 patients in the control group was skin reactive to Parietaria.

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[17] - Rossi RE, Monasterolo G, Coco G, Operti D. Possible Relationship between Systemic Side Effects and Sensitization to rPar j 2 in Allergic Patients Submitted to an Ultra-Rush (20 min) Sublingual Immunotherapy and Selected by Component Resolved Diagnosis. Int Arch Allergy Immunol 2005;138:105-110

BACKGROUND: The pollen of Parietaria spp., a weed of the Urticaceae family, is a major cause of respiratory allergy in the Mediterranean area, where the most common species are Parietaria judaica and Parietaria officinalis. In this study, we evaluated the specific serum IgE-binding profiles to individual P. judaica pollen recombinant major allergen, and Phleum pratense cytoskeletal profilin and a 2-EF-hand calcium-binding allergen homologous to cross-reactive Parietaria pollen allergens, in patients allergic to pollen with positive skin test towards Parietaria spp. extract . METHODS: The present observation included 220 patients from the province of Cuneo, north-west Italy, all suffering from rhino-conjunctivitis and/or asthma selected on the basis of skin test positive to P. judaica extract. The sera were evaluated for specific IgE reactivity to P. judaica pollen major recombinant(r) allergen Par j 2, Phleum pratense pollen allergens rPhl p 7 (2-EF-hand calcium binding protein) and rPhl p 12 (profilin), both identified as cross-reactive Parietaria spp. allergens, using Pharmacia CAP System. Out of 220 patients, 37 patients with IgE reactivity to rPar j 2 and 105 patients sensitized to at least one timothy pollen major allergen (i. e. rPhl p 1, rPhl p 2, natural Phl p 4 and rPhl p 6) were submitted to an ultra-rush protocol of sublingual immunotherapy (SLIT). The occurrence of adverse reactions were evaluated in both groups . RESULTS: All 220 patients with pollinosis and positive in vivo skin prick tests had in vitro positive CAP results to P. judaica natural extract. On the contrary, in these patients the prevalence of Par j 2-specific IgE was only 33.2% (73/220). In fact, 116/220 (52.7%) patients with serum specific IgE to crude Parietaria pollen extract had specific IgE to Phl p 12, 18/220 (8.1%) subjects with specific IgE to rPhl p 12 also exhibited specific IgE to Phl p 7 and 26/220 (11.8%) subjects had specific IgE against rPhl p 7. Particularly, geometric mean (25th-75th percentile) of specific IgE to rPar j 2 were as follows: 2.87 kUA/l (1.005-7.465). Out of 73 patients with specific IgE to rPar j 2, 7 subjects (9.6%) had also specific IgE to rPhl p 7, 12 (16.4%) had specific IgE to rPhl p 12 and 4 (4.1%) patients had specific IgE to both recombinant allergens. Of 37 patients under an ultra-rush protocol of SLIT, 3 subjects (8.1%) experienced generalized urticaria, and 1 of them also had diarrhea 3 h after the last dose of Parietaria judaica extract oral-vaccine administration. On the contrary, no systemic reactions were observed in 105 patients after Phleum pratense extract oral intake after a similar ultra-rush SLIT protocol (p = 0.0046) . CONCLUSIONS: In the light of present findings, allergen extract-based diagnosis, in vivo and in vitro, cannot discriminate allergic patients that are genuinely sensitized to Parietaria spp. major allergens or to other major allergens to which current immunotherapeutic allergy extracts are standardized. Therefore, in vitro component resolved diagnosis is the unique tool to define the disease eliciting molecule(s). Finally, during sublingual immunotherapy, not only the dose of allergen, but also the biochemical characteristic of the major allergen administered may be an important factor in determining possible systemic reactions.

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[18] - Asero R, Mistrello G, Roncarolo D, Amato S. Parietaria Profilin Shows Only Limited Cross-Reactivity with Birch and Grass Profilins. Int Arch Allergy Immunol 2004;133:121-124

BACKGROUND: In patients showing multiple reactivity to seasonal airborne allergens, a skin prick test (SPT) with Parietaria pollen scores frequently negative, suggesting that profilin from Parietaria might not share IgE-binding epitopes with profilin from botanically unrelated airborne allergens. This study investigated the immunologic cross-reactivity between profilins from Parietaria, grass and birch pollen . METHODS: 36 patients hypersensitive to birch profilin, Bet v 2, underwent SPT with Parietaria, and IgE to both whole Parietaria pollen and Phleum profilin were sought in their sera by ELISA. In ELISA inhibition studies, IgE reactivity to Phleum profilin was measured before and after absorption of sera from Parietaria reactors with both whole Parietaria pollen extract, and Par j 1/Par j 2 fraction. Further, their IgE reactivity to whole Parietaria pollen was measured before and after absorption with the Par j 1/Par j 2 fraction . RESULTS: All sera showed IgE reactivity to Phleum profilin. Only 14/36 (39%) patients had a positive SPT to Parietaria and 17/36 (47%) showed IgE to Parietaria. Absorption of sera from Parietaria reactors with whole Parietaria extract caused a marked reduction in IgE reactivity to grass profilin; in contrast, absorption of sera with the Par j 1/Par j 2 fraction did not exert any inhibitory effect on IgE reactivity to grass profilin. Absorption of sera with the Par j 1/Par j 2 fraction markedly reduced IgE reactivity to whole Parietaria extract in 8/9 cases . CONCLUSION: Less than 50% of patients sensitized to birch and grass profilin cross-react to Parietaria profilin. In most cases, cross-reactors are hypersensitive to major, specific Parietaria allergens as well. This findings may be of practical relevance when the prescription of specific immunotherapy is considered.

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[19] - 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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[20] - Colombo P, Bonura A, Costa MA, Izzo V, Passantino R, Locorotondo G, et al. The Allergens of Parietaria. Int Arch Allergy Immunol 2003;130:173-179

Parietaria is a genus of dicotyledonous weeds of the Urticaceae family including several species and its pollen grain is one of the most important allergenic sources in the Mediterranean area. Species belonging to this genus induce IgE responses in approximately 10 million people. Identification of allergens by means of independent strategies suggest that the allergens of the two more common species, Parietaria judaica and Parietaria Officinalis, show molecular weights ranging between 10 and 14 kD and that the allergens of the two extracts are highly cross-reactive. Biochemical analysis and molecular cloning allowed the isolation and immunological characterization of the two major allergens of the P. judaica pollen, Par j 1 and Par j 2. Sequence comparison suggests that the P j major allergens of P. Judaica belong to the nonspecific lipid transfer protein family, and three-dimensional modeling by homology has revealed that both proteins present a very conserved structural motif composed of four alpha-helices. Immunological analysis has shown that Par j 1 and Par j 2 are able to bind most of the P. Judaica-specific IgE and some of their IgE determinants have been mapped. Recombinant Par j 1 and Par j 2 allergens have been shown to possess immunological properties equivalent to their natural counterpart and their availability represents a fundamental tool for the diagnosis and therapy of Parietaria pollen allergy.

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[21] - Gonzalez Romano ML, Gallego MT, Berrens L. Extraordinary stability of IgE-binding Parietaria pollen allergens in relation to chemically bound flavonoids. Mol Immunol 1996;33:1287-1293

It is known that the skin-active and IgE-binding components in Parietaria pollen extracts are not restricted to the predominant protein allergens of M(r) 12000-15000, but are present as well among the naturally occurring constituents of M(r) < 10000. Indeed, the IgE-binding Parietaria pollen components are quite heterogeneous, ranging from high- to low-molecular mass, whereby the IgE-binding epitopes display an unusual chemical stability. Furthermore, the pollen of Parietaria species demonstrably contain a high proportion of flavonoid pigments. Since these pollen grains cannot be collected entirely free from non-pollen plant parts, the usual allergenic extracts of Parietaria encompass both the polyphenolic substrate molecules and the enzyme polyphenoloxidase as ingredients for the oxidative generation of flavonol-protein conjugates during the extraction process. In the present work this is illustrated by spectroscopic analyses of the free and bound flavonoids in Parietaria pollen extracts, as well as of the peptide fragments produced from the allergenic proteins by enzymatic or chemical hydrolysis. None of these relatively harsh treatments had a significant effect on the IgE-binding properties of the allergenic (sub-)components, even though detectable proteins in isoelectric focusing and immunoblotting were lost. It is proposed that the extraordinary stability of IgE-binding Parietaria components over a wide molecular range may be attributed to chromophoric flavonoid side-chains as (parts of) the corresponding B-cell epitopes.

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[22] - Cortes L, Carvalho AL, Todo-Bom A, Faro C, Pires E, Verissimo P. Purification of a novel aminopeptidase from the pollen of Parietaria judaica that alters epithelial integrity and degrades neuropeptides. J Allergy Clin Immunol 2006;118:878-884

BACKGROUND: Parietaria judaica pollen is a common cause of pollinosis in the Mediterranean area . OBJECTIVE: This study sought to purify and characterize the peptidase responsible for the majority of proteolytic activity present in the pollen extract of P judaica, and to investigate its contribution to the allergic response . METHODS: A serial of chromatographic steps was applied to isolate the peptidase from P judaica's pollen, and its biochemical properties were determined. Bioactive peptides present in the airways were incubated with the peptidase, and their degradation was visualized by direct protein sequencing. In addition, we measured the cellular detachment, by methylene blue binding assay, of an airway-derived epithelial cell line (A549) in the presence of the peptidase, and visualized, by Western blot, the degradation of proteins from intercellular junctions . RESULTS: We purified a 98-kDa peptidase from the pollen of P judaica that was classified as an aminopeptidase on the basis of its biochemical properties and internal amino acid sequence. The aminopeptidase was able to degrade bioactive peptides. Moreover, the aminopeptidase caused cellular detachment of A549 cell line and degradation of occludin and E-cadherin . CONCLUSION: Our results suggest that the P judaica aminopeptidase can alter the integrity of the epithelium barrier by degrading occludin as well as E-cadherin. In addition, P judaica aminopeptidase can degrade bioactive peptides, which can exacerbate the overall bronchoconstrictive effect detected in asthmatic lungs. CLINICAL IMPLICATIONS: The novel aminopeptidase described here could constitute a relevant therapeutic target in the treatment of allergic disorders induced by the pollen of P judaica.

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[23] - Corbi AL, Ley V, Sanchez-Madrid F, Carreira J. Isolation of the major IgE-binding protein from Parietaria judaica pollen using monoclonal antibodies. Mol Immunol 1985;22:1081-1089

Allergen molecules from Parietaria judaica pollen, a widely distributed allergy inducer in Southern and Western Europe, have been studied using specific monoclonal antibodies (MAbs). MAbs against IgE-binding components were selected in a 4-step radioimmunoassay. Three different MAbs (AC/1.1, AC/7.1 and AC/15.1) were obtained which recognized epitope(s) located on a polypeptide of 10 Kd (Pj10). This polypeptide displayed the highest IgE-binding ability under either native or SDS-denatured conditions, as determined by immunoadsorption and immunodetection after SDS-PAGE, respectively. The Pj10-containing allergen, purified on an AC/1.1 MAb-Sepharose column, was able to inhibit most of the binding of specific IgE to the pollen extract coupled to paper discs in an inhibition radioallergosorbent test (RAST). The affinity-purified allergen exhibited the same immunoelectrophoretic behaviour as the native allergen.

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[24] - Corbi AL, Carreira J. Identification and characterization of Parietaria judaica allergens. Int Arch Allergy Appl Immunol 1984;74:318-323

Allergens of Parietaria judaica pollen extract have been identified and characterized biochemically. Two main allergenic components, A1 and A2, have been found by crossed-radioimmunoelectrophoresis and demonstrated to be spread in a wide range of pH. Immunoblotting studies revealed that at least eight SDS-denatured polypeptides show IgE-binding activity. The one exhibiting the highest allergenic activity, named Pj10 (MW 10,000 daltons) was found in all the fractions when the pollen extract was fractionated by chromatofocusing. Bidimensional electrophoretic analysis suggested that Pj10 either can form homopolymeric proteins of different molecular weights or can be associated to a number of proteins by disulfide bridges. Furthermore, Pj10 is the main molecular structure with IgE-binding activity in the two allergenic components A1 and A2 defined by immunological criteria

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[25] - Ayuso R, Polo F, Carreira J. Purification of Par j I, the major allergen of Parietaria judaica pollen. Mol Immunol 1988;25:49-56

A component of Parietaria judaica pollen extract, previously identified as the major allergen, then reported as Pj10 and hereafter denominated Par j I has been isolated by a combination of 65% ammonium sulphate salt precipitation and gel filtration and an Ultrogel AcA54 column. The purified allergen appeared essentially homogeneous on gel filtration HPLC The mol. wt of Par j I was estimated by electrophoretic and chromatographic techniques. All results gave values in a range from 13 K to 25 K. Analysis in SDS-PAGE under reducing conditions revealed a broad band corresponding to a mol. wt of 10 K, which retained allergenicity when tested with a patients serum pool. CIE and CRIE patterns of the isolated Par j I displayed the two precipitating lines already reported as those exhibiting the highest IgE-binding ability. Par j I showed a specific allergenic activity about 10-fold higher than that of the whole extract and was demonstrated to be the major allergen of Parietaria judaica as assessed in 25 sensitive human sera.

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[26] - Colombo P, Bonura A, Costa MA, Izzo V, Passantino R, Locorotondo G, et al. The Allergens of Parietaria. Int Arch Allergy Immunol 2003;130:173-179

Parietaria is a genus of dicotyledonous weeds of the Urticaceae family including several species and its pollen grain is one of the most important allergenic sources in the Mediterranean area. Species belonging to this genus induce IgE responses in approximately 10 million people. Identification of allergens by means of independent strategies suggest that the allergens of the two more common species, Parietaria judaica and Parietaria Officinalis, show molecular weights ranging between 10 and 14 kD and that the allergens of the two extracts are highly cross-reactive. Biochemical analysis and molecular cloning allowed the isolation and immunological characterization of the two major allergens of the P. judaica pollen, Par j 1 and Par j 2. Sequence comparison suggests that the P j major allergens of P. Judaica belong to the nonspecific lipid transfer protein family, and three-dimensional modeling by homology has revealed that both proteins present a very conserved structural motif composed of four alpha-helices. Immunological analysis has shown that Par j 1 and Par j 2 are able to bind most of the P. Judaica-specific IgE and some of their IgE determinants have been mapped. Recombinant Par j 1 and Par j 2 allergens have been shown to possess immunological properties equivalent to their natural counterpart and their availability represents a fundamental tool for the diagnosis and therapy of Parietaria pollen allergy.

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[27] - Asturias JA, Gomez-Bayon N, Eseverri JL, Martinez A. Par j 1 and Par j 2, the major allergens from Parietaria judaica pollen, have similar IgE epitopes. Clin Exp Allergy 2003;33:518-524

BACKGROUND: Parietaria judaica is the main cause of allergy in Mediterranean countries. The major allergens from P. judaica pollen, Par j 1 and Par j 2, have amino acid sequence identity of 45% and contain eight cysteine residues involved in disulphide bonds that compact the structure . OBJECTIVE: The aim of the study was to identify IgE-binding epitopes on Par j 1 and Par j 2, the major allergens from P. judaica pollen and correlate them with the three-dimensional structure of the proteins . METHODS: Overlapping peptides representing the complete length of Par j 1 and Par j 2 were synthesized on a cellulose-derivatized membrane. Sera from 17 P. judaica-allergenic patients were used to identify IgE-binding epitopes. Homology modelling of the three dimensional structure of both allergens was generated using the Swiss-Model software on the basis of previously reported crystal structures . RESULTS: Five and eight IgE-binding epitopes were identified on Par j 1 and Par j 2 allergens, respectively. Both proteins belonged to the non-specific lipid transfer proteins (ns-LTP) family and therefore a three-dimensional model of both allergens was constructed on the basis of the maize ns-LTP crystal structure. Par j 1 and Par j 2 allergens have three similar allergenic epitopes with high homology and identical conformation . CONCLUSION: Three similar IgE-epitopes of major allergens from P. judaica have been described. They could be good candidates for designing of IgE haptens as therapeutic agents with reduced anaphylactic side-effects or for developing hypoallergenic variants of these major allergens.

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[28] - Duro G, Colombo P, Costa MA, Izzo V, Porcasi R, Di Fiore R, et al. cDNA cloning, sequence analysis and allergological characterization of Par j 2.0101, a new major allergen of the Parietaria judaica pollen. FEBS Lett 1996;399:295-298

A clone (P2) coding for an allergen of Parietaria judaica (Pj) pollen has been isolated and sequenced from a cDNA library in lambda ZAP using a pool of 23 sera from Pj-allergic patients. The clone contained an insert of 622 nucleotides with an open reading frame of 133 amino acids (aa) and a putative signal peptide of 31 aa giving a deduced mature processed protein of 102 aa with a molecular mass of 11344 Da. The expressed recombinant protein, named rPar j 2.0101, was a major allergen since it reacted with IgE of 82% (23/28) of the sera of Pj- allergic subjects analyzed. It was shown to be a new allergen since (i) the amino acid sequence homology with the already reported recombinant allergen Par j 1.0101 was 45% and (ii) there was no cross-inhibition between rPar j 2.0101 and rPar j 1.0101. In addition, rPar j 2.0101 inhibited 35% of the specific IgE for 10-14 kDa native allergens and preincubation of sera from Pj-allergic patients with both rPar j 2.0101 and rPar j 1.0101 fully abolished the IgE recognition of the 10-14 kDa native allergen region, suggesting that these two allergens contributed to the region

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[29] - Ayuso R, Carreira J, Polo F. Quantitation of the major allergen of several Parietaria pollens by an anti-Par 1 monoclonal antibody-based ELISA. Analysis of crossreactivity among purified Par j 1, Par o 1 and Par m 1 allergens. Clin Exp Allergy 1995;25:993-999

Plants of the genus Parietaria, Urticaceae family, represent a major cause of pollinosis in the Mediterranean area. Different Parietaria species crossreact to a great extent, but studies on the crossreactivity among the major allergens of these pollens have not been carried out so far. OBJECTIVE: To develop an immunochemical method to quantify the major Parietaria judaica allergen, Par j 1, as well as to verify the presence of Par j 1-like proteins in different Urticaceae pollens. These proteins would be purified in order to study the cross-reactivity among them. METHODS: Immunoaffinity chromatography with a monoclonal antibody, solid-phase enzyme-linked immunoassays and SDS-PAGE. RESULTS: A monoclonal antibody-based ELISA for the quantification of Par j 1 has been developed. The assay has a sensitivity of 0.2 ng/mL and shows a high correlation with the allergenic activity of P. judaica extracts determined by radioallergosorbent assay (RAST) inhibition. By means of this assay, proteins homologous to Par j 1 were detected in P. officinalis and P. mauritanica. These proteins (Par o 1 and Par m 1, respectively) were purified by affinity chromatography using the same monoclonal antibody employed in the ELISA. Crossed-inhibition experiments demonstrated that Par j 1, Par o 1, and Par m 1, competed for the binding of specific IgE from a P. judaica-sensitive patients serum pool. CONCLUSION: The results here described suggest that shared allergenic epitopes are present in the three main allergens investigated, which may simplify the diagnosis and therapy for Parietaria allergy.

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[30] - Mucci N, Liberatore P, Federico R, Forlani F, Di Felice G, Afferni C, et al. Role of carbohydrate moieties in cross-reactivity between different components of Parietaria judaica pollen extract. Allergy 1992;47:424-430

Cross-reactivity between the different components in Parietaria judaica pollen extract has been investigated by polyclonal as well as monoclonal antibodies before and after chemical deglycosylation obtained by trifluoromethanesulphonic acid (TFMS) treatment of the extract. In western blotting a polyclonal rabbit antiserum, obtained by injecting purified Par j I, was able to recognise many components of the native extract. However, its reactivity was restricted, after chemical deglycosylation of the extract, to the major allergen alone, indicating that its cross-reactivity was due to sugar moieties. Moreover, out of several monoclonal antibodies raised by injecting the whole Parietaria judaica extract, one (1A4/2F8) was also able in western blotting to recognise an epitope shared by many components of the extract except the major allergen Par j I. However, in this case the broad reactivity of the antibody was not affected by the deglycosylating procedure. When the reactivity of Parietaria judaica extract was tested before and after sugar removal, against specific IgE from a pool of patient sera, no differences could be demonstrated, thus indicating that carbohydrates are not strongly involved in the binding of Parietaria judaica-specific IgE. The results indicate that both proteic and carbohydratic cross-reactive epitopes are shared by many components of Parietaria judaica pollen extract.

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[31] - Ayuso R, Carreira J, Lombardero M, Duffort O, Peris A, Basomba A, et al. Isolation by mAb based affinity chromatography of two Par j I isoallergens. Comparison of their physicochemical, immunochemical and allergenic properties. Mol Immunol 1993;30:1347-1354

We report the identification and separation of two isoallergen components of Par j I, the major allergen from Parietaria judaica pollen. First, electrophoretic conditions for consistently separating both isoforms in an SDS-PAGE system were established, and mol. wt values of 13,000 (isoallergen IA) and 10,500 (isoallergen IB) were estimated. Immunoblot, after SDS-PAGE experiments, with individual P. judaica-sensitive human sera revealed a slightly different IgE-binding pattern for each isoallergen. Four anti-Par j I mAbs were obtained from BALB/c mice immunized with a purified Par j I preparation comprising IA and IB isoallergens. Three mAbs were directed to an epitope shared by both isoallergens, and the fourth one recognized specifically one epitope on Par j IB. Dot-blot experiments with the deglycosylated allergen showed that the mAbs did not recognize the carbohydrate prosthetic group of the molecules. Affinity chromatography using the mAbs allowed the separation of the isoallergens that retained their IgE-binding ability after the purification process. Amino acid composition analyses and partial N-terminal sequencing demonstrated an extensive homology and also the existence of some structural differences between Par j I isoallergens, which is in agreement with the high, but not complete, cross-reactivity observed in competition ELISA experiments. Finally, skin prick tests performed on 28 P. judaica-sensitive patients showed that all of them recognized both isoforms and that allergenic epitopes present in Par j IA and IB are responsible for most of the allergenic activity of the whole extract.

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[32] - Polo F, Ayuso R, Carreira J. Studies on the relationship between structure and IgE-binding ability of Parietaria judaica allergen I. Mol Immunol 1991;28:169-175

The main allergen of Parietaria judaica pollen, Par j I, is a glycopolypeptide with mol. wt about 10,000. It shows a considerable charge heterogeneity which is mostly due to the carbohydrate prosthetic groups, since treatment with trifluoromethanesulfonic acid yielded a deglycosylated protein with 8500 mol. wt that displayed only a few bands on IEF, in a narrow pH-region around 5.0. Deglycosylated Par j I exhibited a specific allergenic activity slightly lower than that of native Par j I; however, no allergenic determinants should be located on the sugar moiety since both native and deglycosylated Par j I inhibited up to a similar extent the binding of specific human IgE to P. judaica-coated wells in ELISA. The decrease of specific allergenic activity following deglycosylation could be ascribed to conformational changes evidenced by CD experiments. On the other hand, fluorescence spectroscopy showed that Par j I bears unidentified yellow-brown chromophores strongly linked to the polypeptide chain. These chromophores were not removed by TFMS treatment. Finally, reduction and alkylation caused the complete loss of allergenic activity, showing that disulphide bridges are essential for the IgE-binding ability of Par j I.

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[33] - Amoresano A, Pucci P, Duro G, Colombo P, Costa MA, Izzo V, et al. Assignment of disulphide bridges in Par j 2.0101, a major allergen of Parietaria judaica pollen. Biol Chem 2003;384:1165-1172

Par j 2.0101, a major allergen of the Parietaria judaica pollen, was expressed in E. coli, purified to homogeneity and fully characterised both at the structural and the functional level. The recombinant rPar j 2.0101 protein showed an allergenic activity in histamine release, skin prick tests and capacity to bind IgE, almost identical to that of the native allergens purified from aqueous pollen extract. The complete pattern of S-S bridges of rPar j 2.0101 was determined by enzymatic digestion with endoproteinase Lys-C followed by mass spectrometric analysis of the resulting peptide mixtures. The eight cysteines occurring in the allergenic protein were found to be paired into the following four disulphides: Cys35-Cys83, Cys45-Cys60, Cys61-Cys106 and Cys81-Cys121. This structural information probes Par j 2.0101 to attain a 3-D fold consistent with that of the non-specific lipid transfer protein (ns-LTP) family and it represents an effective molecular basis to develop modified antigens by selective site-directed mutagenesis for immunotherapy.

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[34] - Lombardero M, Garcia-Sellés FJ, Polo F, Jimeno L, Chamorro MJ, Garcia-Casado G, et al. Prevalence of sensitization to Artemisia allergens Art v 1, Art v 3 and Art v 60 kDa. Cross-reactivity among Art v 3 and other relevant lipid-transfer protein allergens. Clin Exp Allergy 2004;34:1415-1421

BACKGROUND: Artemisia vulgaris is a widespread weed in the Mediterranean area and several allergens have been detected in its pollen. One of them, Art v 3, belongs to the lipid-transfer protein (LTP) family and its prevalence in Artemisia-sensitized patients or its relationship with other LTP allergens is not clear . OBJECTIVE: To assess the pattern of sensitization to an array of mugwort allergens in a Mediterranean population, and to study the cross-reactivity of Art v 3 with Pru p 3 and Par j 1, relevant LTP allergens in the area . METHODS: Skin prick test was performed with whole extracts (A. vulgaris, Parietaria judaica and peach) and pure natural allergens Art v 1, Art v 3, Art v 60 kDa and Par j 1 in 24 mugwort-allergic patients from a Mediterranean area. In vitro assays included measurement of specific IgE and ELISA inhibition among LTP allergens . RESULTS: The three Artemisia allergens elicited a positive skin response in 70-80% of the patients. Seven patients were clearly sensitized to Par j 1 and 11 to Pru p 3. There was no correlation between Par j 1 and Pru p 3 sensitization, but a highly significant correlation was found between peach extract and Art v 3 as regards the skin response. No IgE cross-reactivity was observed between Art v 3/Par j 1 or Pru p 3/Par j 1. In contrast, Art v 3 significantly inhibited the binding to Pru p 3 of IgE from three patients' sera out of six studied, but Pru p 3 was not able to inhibit the IgE binding to Art v 3 . CONCLUSION: Art v 3 is a major mugwort allergen and in some patients with IgE to both Art v 3 and Pru p 3, Art v 3 behaves as the primary sensitizing agent.

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[35] - Vassilopoulou E, Zuidmeer L, Akkerdaas J, Tassios I, Rigby NR, Mills ENC, et al. Severe Immediate Allergic Reactions to Grapes: Part of a Lipid Transfer Protein-Associated Clinical Syndrome. Int Arch Allergy Immunol 2007;143:92-102

BACKGROUND: Grape allergy is considered rare; grape lipid transfer protein (LTP; Vit v 1), an endochitinase and a thaumatin-like protein (TLP) have been reported as grape allergens. A considerable number of patients have referred to our department for severe reactions to grapes, and several IgE binding proteins were detected . OBJECTIVES: The aim of this study was to identify and characterise the allergens involved in severe allergic reactions to grapes and describe the population in which they occur . METHODS: Patients with reported severe allergic reactions to grapes (n = 37) are described. Grape allergens were purified/fractionated by a combination of chromatographic techniques, identified by proteomic analysis and biochemically characterised. Immunoreactivity was assessed by blot (inhibitions) and RAST (inhibitions), and skin prick tests were performed with the isolated allergens . RESULTS: All subjects were polyallergic, sensitised and reactive to several additional foods and pollen. All patients were sensitised to grape LTP. A 28-kDa expansin, a 37.5-kDa polygalacturonase-inhibiting protein, a 39-kDa beta-1,3-glucanase and a 60-kDa protein were identified as minor grape allergens. Endochitinase and TLP did not play a role. Inhibition experiments revealed the possible cross-reactive role of LTP for clinical sensitivities to other LTP-containing plant foods, but also the involvement of cross-reactive carbohydrate determinants of minor allergens in IgE cross-reactivity . CONCLUSIONS: LTP is the major grape allergen, while additional minor allergens may contribute to clinical reactivity. Severe grape allergy presents in atopic patients who frequently react to other LTP-containing, plant-derived foods. The 'LTP syndrome' is the appropriate term to describe this condition.

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[36] - Pastorello EA, Pravettoni V, Farioli L, Rivolta F, Conti A, Ispano M, et al. Hypersensitivity to mugwort (Artemisia vulgaris) in patients with peach allergy is due to a common lipid transfer protein allergen and is often without clinical expression. J Allergy Clin Immunol 2002;110:310-317

Background: The observation of mugwort-specific IgE antibodies in patients with peach allergy suggests that mugwort sensitization might play a role in sensitization to peach. Objective: We sought to study the clinical manifestations of mugwort hypersensitivity in patients with peach allergy, identify the common allergens, and evaluate their IgE crossreactivity. Methods: Patients with oral allergy syndrome for peach and specific IgE antibodies to mugwort were investigated for respiratory symptoms during the mugwort season. Peach and mugwort allergens were identified by means of SDS-PAGE and IgE immunoblotting. Immunoblotting inhibition experiments were done to study cross-reactivity between peach and mugwort and other pollens. Results: Seventeen patients were studied, 10 with no seasonal respiratory symptoms and 7 with clear late summer respiratory symptoms. In IgE immunoblotting the 10 asymptomatic patients reacted only to a 9-kd allergen of both mugwort and peach, whereas the 7 patients with pollinosis reacted to other allergens. Ten patients with mugwort allergy, no history of allergy to peach, and negative results for peach-specific IgE antibodies were also studied. The mugwort 9-kd protein was identified as a lipid transfer protein (LTP) homologous to peach LTP. Immunoblotting inhibition showed that IgE binding to the peach 9-kd band was totally inhibited by 4 µg of peach LTP but only by 400 µg of mugwort LTP, whereas 4 µg of both mugwort and peach LTP totally inhibited the mugwort immunoblotting. The results were similar with other pollens. Conclusions: Patients sensitized only to the 9-kd LTP of mugwort do not present hay fever symptoms, and this sensitization is a consequence of the peach sensitization.

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[37] - Suárez-Cervera M, Vega-Maray A, Rodríguez-Rajo F, Asturias J, Fernández-González D, Jato V et al. Lipid transfer protein allergy. An approach to the knowledge of pollen and allergen diversity. Allergy 2009;64(Suppl. 90):508

Background: The aim of the present study is to localize non-specific lipid transfer proteins (nsLTPs), obtained from Rosaceae, Prunus persica, in pollen grains of taxonomically distant plants, such as Iridaceae, Aristea latifolia, Platanaceae, Platanus acerifolia and Urticaceae, Parietaria judaica, in order to compare pollen and nsLTP diversity. Methods: A combination of transmission electron microscopy, immunocytochemical techniques, and rabbit specific antiserum against peach nsLTPs (Pru p 3) were used. Results: Abundant labelling to Pru p 3-like proteins was observed in the cytoplasm, walls and pollenkitt of A. latifolia pollen grains. The labelling was less intense in the cytoplasm and walls of P. acerifolia. Immuno-stained gold particles were associated with the vacuoles, lipid inclusions, endoplasmic reticulum and Golgi complex. No significant labelling was found in the P. judaica pollen grains incubated with anti-Pru p 3 polyclonal antibodies. This was expected because Par j 1-2 and Pru p 3 nsLTPs have strong sequence divergence. Conclusions: These results show distinct patterns of distribution in the nsLTPs of different pollen grain species. Consequently, no taxonomic relationship between pollen grains and nsLTPs could be established. This should be kept in mind for the cross-reactivity processes and allergen extract preparation in terms of botanical groups. On the other hand, the localization of nsLTPs in the extracellular pollenkitt provides evidence that this substance is involved in the traffic of pollen defence proteins. Moreover, the lipid traffic pathways proposed for LTPs in the cytoplasm of other cellular tissues could be applied to the living cells of pollen grains.

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[38] - Mari A, Artale A, Bonura A, Scala E, Colombo P. Par j 2 as a parietaria sensitisation marker allergen: definition of its use in epidemiology and diagnosis. Allergy Clin Immunol Int 2005;17(Suppl. 1):205

Several allergens have been identifed, characterised and cloned from Parietaria pollen. Par j 1 and Par j 2 represent allergenic molecules recognised by patients' IgE who are usually not sensitised to other pollen sources. The recombinant Par j 2 (rPar j 2) is now available for in vitro testing (Pharmacia, Sweden). We examined a cohort of subject with pollinosis symptoms. Subjects were from urban and non-urban areas, with different exposure levels to Parietaria. Patients underwent skin test (SPT) with a previously defined panel of pollen allergenic extracts and serum samples were obtained. IgE reactivity to Parietaria judaica extract (PjE) was compared to rPar j 2 reactivity. A cohort of 2993 pollinosis subject were identified, 37.1 % had a positive SPT to a PjE. IgE to PjE were detected in 96.3 % of 709 blood samples. rPar j 2 IgE positive samples were 82.2% of 573 PjE IgE positive samples. Differences were recorded when different patient subsets were considered. 92.5 % of Parietaria mono-sensitised subjects (n= 80); SPT and IgE positive had IgE to rParj 2. Similar results were recorded when Parietaria was associated to other pollens (92%; n=289), whereas only 64.2% were positive in the group of multiple pollen sensitised subjects (MPS; n=204). Differences were recorded in geographical distribution of rPar j 2 reactive subjects. A relatively small area with statistically significant lower prevalence rPar j 2 IgE positive subjects was identified when native subjects were considered. Correlating IgE results, PjE vs rPar j 2, the best result was achieved in the mono-sensitised and oligo-sensitised cohort again if compared to the MPS patient subset. When results from mono-sensitised subjects were plotted, three further subgroups were identified. A first group with almost overlapping values for both PjE and rPar j 2, a second with PjE values greater than rPar j 2 values, and a third group with the opposite finding. Representative sera from these three subgroups have been tested for both rPar j 1 and 2. All reacted with both. rPar j 2 can be assumed as a specific marker for exposure and sensitisation to Parietaria pollen. rPar j 2 evaluation allows both a more precise geographical mapping and for identification of Parietaria sensitised subjects within MPS subset. Further studies are needed to define whether a single molecule (rPar j 2) is suitable for both diagnosis and future molecule-based immunotherapy.

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[39] - Asturias JA, Gómez-Bayón N, Eseverri JL, Martínez A. IgE epitope analysis of Par j 1 and Par j 2, the major allergens of Parietaria judaica. EAACI 21th Congress, Naples, 1-5 June, 2002, Poster n°922

Parietaria judaica pollen is the main cause of allergy in the Mediterranean area with a prevalence of 60-80% in Italy and Greece and 25-40% in Spain and Southern France. This pollen contains at least nine allergens, and two of them, named Par j 1 and Par j 2, have been cloned and sequenced. Par j 1 is a 14.7 kDa protein with a prevalence of 95% among P. judaica allergic patients. Par j 2 is a protein of 11.3 kDa which reacted with IgE of 82% of the sera of P. judaica allergic subjects. Both proteins belong to the family of protein refereed to as non-specific lipid transfer proteins (ns-LTP). In the present study, we used overlapping synthetic peptides to identify major IgE-binding epitopes of Par j 1 and Par j 2 in patients with Parietaria pollen allergy. Serum samples were obtained from 17 patients with P. judaica allergy. All patients were chosen based on their clinical history, skin prick test reactivity, specific IgE class > 2, and immunoblot reactivity to 10-14 kDa Par j 1/2. Individual peptides were synthesized on a derivatized cellulose membrane using Fmoc chemistry (SPOTs). Homology modeling of the three-dimensional structure of Par j 1 and Par j 2 was generated using the Swiss-Model server. Six individual IgE-binding epitopes, of 6-10 residues long, were identified in Par j 1 corresponding to regions: Q19-P25, C30-G38, T42-A49, P72-D79, L82-C91, and P118-E125, and named epitopes 1-I to 1-VI, respectively. Eight overlapping IgE-binding epitopes was detected in Par j 2 corresponding to regions: C14-E21, V18-K27, C30-E39, E38-Q45, K46-R55, C52-I60, K73-T80, and S91-F98 and named epitopes 2-I to 2-VIII, respectively. There were no obvious amino acid sequence motifs shared by all the IgE-binding epitopes. The lineal IgE epitopes described were represented on the 3-D structure of both allergens based on the maize ns-LTP. Reactive regions fitted with the second and third a-helix and turns 1, 2 and 4. Amino acid sequence comparisons of these IgE reactive regions showed three epitopes with high homology in both allergens. The availability of three-dimensional model in combination with the known sequences of the IgE-binding sites allows the design of monovalent allergen fragments (haptens) which would not cross-link effector-cell-bound IgE and, therefore, could be used for local blocking therapy as well as for active immunization.

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[40] - Bonura A, Amoroso S, Locorotondo G, di Felice G, Tinghino R, Geraci D, et al. Hypoallergenic Variants of the Parietaria judaica Major Allergen Par j 1: A Member of the Non-Specific Lipid Transfer Protein Plant Family. Int Arch Allergy Immunol 2001;126:32-40

Background: Par j 1 represents a major allergenic component of Parietaria judaica (Pj) pollen, since it is able to induce an immunoglobulin E (IgE) response in 95% of Pj-allergic patients. It belongs to the non-specific lipid transfer protein family, sharing with them a common three-dimensional structure. Methods: Disulphide bond variants of the recombinant Par j 1 (rPar j 1) allergen were generated by site-directed mutagenesis, and the immunological activity of rPar j 1 and its conformational mutants was compared with the use of the skin prick test (SPT). The ability to bind IgE antibodies was evaluated by Western blot, ELISA and ELISA inhibition. T cell reactivity was measured by peripheral blood mononuclear cell proliferation assay. Results: The disruption of Cys14-Cys29 and Cys30-Cys75 bridging (PjA mutant) caused the loss of the majority of specific IgE-binding activity. Additional disruption of the Cys4-Cys52 bridge (PjC mutant) and the latter Cys50-Cys91 bridge (PjD mutant) led to the abolition of IgE-binding activity. On the SPT, PjB (lacking the Cys4-Cys52 and Cys50-Cys91 bridges) was still capable of triggering a type I hypersensitive reaction in 9 out of 10 patients, and PjA in 3 out of 10 patients, while PjC and PjD did not show any SPT reactivity. All the mutants preserved their T cell reactivity. Conclusion: Recombinant hypoallergenic variants of the rPar j 1 allergen described herein may represent a useful tool for improved immunotherapy.

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[41] - Costa MA, Colombo P, Izzo V, Kennedy H, Venturella S, Cocchiara R, et al. cDNA cloning, expression and primary structure of Par j I, a major allergen of Parietaria judaica pollen. FEBS Lett 1994;341:182-186

A 659 bp cDNA clone** coding for an allergen of Pj pollen has been isolated from a lambda gt11 library, and its DNA sequence determined. The cDNA insert showed an open reading frame of 429 bp coding for an allergenic protein of 14,866 Da and a deduced amino acid sequence containing 143 residues. The expressed recombinant protein represented the major allergen Par jI since it reacted with 95% of the sera from Pj- allergic patients (n = 22) and with two Par jI-specific monoclonal antibodies. No similarity with other known DNA and protein sequences has been detected

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[42] - Duro G, Colombo P, Costa MA, Izzo V, Porcasi R, Di Fiore R, et al. cDNA cloning, sequence analysis and allergological characterization of Par j 2.0101, a new major allergen of the Parietaria judaica pollen. FEBS Lett 1996;399:295-298

A clone (P2) coding for an allergen of Parietaria judaica (Pj) pollen has been isolated and sequenced from a cDNA library in lambda ZAP using a pool of 23 sera from Pj-allergic patients. The clone contained an insert of 622 nucleotides with an open reading frame of 133 amino acids (aa) and a putative signal peptide of 31 aa giving a deduced mature processed protein of 102 aa with a molecular mass of 11344 Da. The expressed recombinant protein, named rPar j 2.0101, was a major allergen since it reacted with IgE of 82% (23/28) of the sera of Pj- allergic subjects analyzed. It was shown to be a new allergen since (i) the amino acid sequence homology with the already reported recombinant allergen Par j 1.0101 was 45% and (ii) there was no cross-inhibition between rPar j 2.0101 and rPar j 1.0101. In addition, rPar j 2.0101 inhibited 35% of the specific IgE for 10-14 kDa native allergens and preincubation of sera from Pj-allergic patients with both rPar j 2.0101 and rPar j 1.0101 fully abolished the IgE recognition of the 10-14 kDa native allergen region, suggesting that these two allergens contributed to the region

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[43] - Mari A, Artale A, Bonura A, Scala E, Colombo P. Par j 2 as a parietaria sensitisation marker allergen: definition of its use in epidemiology and diagnosis. Allergy Clin Immunol Int 2005;17(Suppl. 1):205

Several allergens have been identifed, characterised and cloned from Parietaria pollen. Par j 1 and Par j 2 represent allergenic molecules recognised by patients' IgE who are usually not sensitised to other pollen sources. The recombinant Par j 2 (rPar j 2) is now available for in vitro testing (Pharmacia, Sweden). We examined a cohort of subject with pollinosis symptoms. Subjects were from urban and non-urban areas, with different exposure levels to Parietaria. Patients underwent skin test (SPT) with a previously defined panel of pollen allergenic extracts and serum samples were obtained. IgE reactivity to Parietaria judaica extract (PjE) was compared to rPar j 2 reactivity. A cohort of 2993 pollinosis subject were identified, 37.1 % had a positive SPT to a PjE. IgE to PjE were detected in 96.3 % of 709 blood samples. rPar j 2 IgE positive samples were 82.2% of 573 PjE IgE positive samples. Differences were recorded when different patient subsets were considered. 92.5 % of Parietaria mono-sensitised subjects (n= 80); SPT and IgE positive had IgE to rParj 2. Similar results were recorded when Parietaria was associated to other pollens (92%; n=289), whereas only 64.2% were positive in the group of multiple pollen sensitised subjects (MPS; n=204). Differences were recorded in geographical distribution of rPar j 2 reactive subjects. A relatively small area with statistically significant lower prevalence rPar j 2 IgE positive subjects was identified when native subjects were considered. Correlating IgE results, PjE vs rPar j 2, the best result was achieved in the mono-sensitised and oligo-sensitised cohort again if compared to the MPS patient subset. When results from mono-sensitised subjects were plotted, three further subgroups were identified. A first group with almost overlapping values for both PjE and rPar j 2, a second with PjE values greater than rPar j 2 values, and a third group with the opposite finding. Representative sera from these three subgroups have been tested for both rPar j 1 and 2. All reacted with both. rPar j 2 can be assumed as a specific marker for exposure and sensitisation to Parietaria pollen. rPar j 2 evaluation allows both a more precise geographical mapping and for identification of Parietaria sensitised subjects within MPS subset. Further studies are needed to define whether a single molecule (rPar j 2) is suitable for both diagnosis and future molecule-based immunotherapy.

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[44] - Rossi RE, Monasterolo G, Coco G, Operti D. Possible Relationship between Systemic Side Effects and Sensitization to rPar j 2 in Allergic Patients Submitted to an Ultra-Rush (20 min) Sublingual Immunotherapy and Selected by Component Resolved Diagnosis. Int Arch Allergy Immunol 2005;138:105-110

BACKGROUND: The pollen of Parietaria spp., a weed of the Urticaceae family, is a major cause of respiratory allergy in the Mediterranean area, where the most common species are Parietaria judaica and Parietaria officinalis. In this study, we evaluated the specific serum IgE-binding profiles to individual P. judaica pollen recombinant major allergen, and Phleum pratense cytoskeletal profilin and a 2-EF-hand calcium-binding allergen homologous to cross-reactive Parietaria pollen allergens, in patients allergic to pollen with positive skin test towards Parietaria spp. extract . METHODS: The present observation included 220 patients from the province of Cuneo, north-west Italy, all suffering from rhino-conjunctivitis and/or asthma selected on the basis of skin test positive to P. judaica extract. The sera were evaluated for specific IgE reactivity to P. judaica pollen major recombinant(r) allergen Par j 2, Phleum pratense pollen allergens rPhl p 7 (2-EF-hand calcium binding protein) and rPhl p 12 (profilin), both identified as cross-reactive Parietaria spp. allergens, using Pharmacia CAP System. Out of 220 patients, 37 patients with IgE reactivity to rPar j 2 and 105 patients sensitized to at least one timothy pollen major allergen (i. e. rPhl p 1, rPhl p 2, natural Phl p 4 and rPhl p 6) were submitted to an ultra-rush protocol of sublingual immunotherapy (SLIT). The occurrence of adverse reactions were evaluated in both groups . RESULTS: All 220 patients with pollinosis and positive in vivo skin prick tests had in vitro positive CAP results to P. judaica natural extract. On the contrary, in these patients the prevalence of Par j 2-specific IgE was only 33.2% (73/220). In fact, 116/220 (52.7%) patients with serum specific IgE to crude Parietaria pollen extract had specific IgE to Phl p 12, 18/220 (8.1%) subjects with specific IgE to rPhl p 12 also exhibited specific IgE to Phl p 7 and 26/220 (11.8%) subjects had specific IgE against rPhl p 7. Particularly, geometric mean (25th-75th percentile) of specific IgE to rPar j 2 were as follows: 2.87 kUA/l (1.005-7.465). Out of 73 patients with specific IgE to rPar j 2, 7 subjects (9.6%) had also specific IgE to rPhl p 7, 12 (16.4%) had specific IgE to rPhl p 12 and 4 (4.1%) patients had specific IgE to both recombinant allergens. Of 37 patients under an ultra-rush protocol of SLIT, 3 subjects (8.1%) experienced generalized urticaria, and 1 of them also had diarrhea 3 h after the last dose of Parietaria judaica extract oral-vaccine administration. On the contrary, no systemic reactions were observed in 105 patients after Phleum pratense extract oral intake after a similar ultra-rush SLIT protocol (p = 0.0046) . CONCLUSIONS: In the light of present findings, allergen extract-based diagnosis, in vivo and in vitro, cannot discriminate allergic patients that are genuinely sensitized to Parietaria spp. major allergens or to other major allergens to which current immunotherapeutic allergy extracts are standardized. Therefore, in vitro component resolved diagnosis is the unique tool to define the disease eliciting molecule(s). Finally, during sublingual immunotherapy, not only the dose of allergen, but also the biochemical characteristic of the major allergen administered may be an important factor in determining possible systemic reactions.

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[45] - Kanny G, Astier C, Petit N, Volpé JS. Allergie alimentaire au fenugrec: à propos de deux observations. Alim'Inter 2009;14:108-112

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[46] - Mari A, Artale A, Bonura A, Scala E, Colombo P. Par j 2 as a parietaria sensitisation marker allergen: definition of its use in epidemiology and diagnosis. Allergy Clin Immunol Int 2005;17(Suppl. 1):205

Several allergens have been identifed, characterised and cloned from Parietaria pollen. Par j 1 and Par j 2 represent allergenic molecules recognised by patients' IgE who are usually not sensitised to other pollen sources. The recombinant Par j 2 (rPar j 2) is now available for in vitro testing (Pharmacia, Sweden). We examined a cohort of subject with pollinosis symptoms. Subjects were from urban and non-urban areas, with different exposure levels to Parietaria. Patients underwent skin test (SPT) with a previously defined panel of pollen allergenic extracts and serum samples were obtained. IgE reactivity to Parietaria judaica extract (PjE) was compared to rPar j 2 reactivity. A cohort of 2993 pollinosis subject were identified, 37.1 % had a positive SPT to a PjE. IgE to PjE were detected in 96.3 % of 709 blood samples. rPar j 2 IgE positive samples were 82.2% of 573 PjE IgE positive samples. Differences were recorded when different patient subsets were considered. 92.5 % of Parietaria mono-sensitised subjects (n= 80); SPT and IgE positive had IgE to rParj 2. Similar results were recorded when Parietaria was associated to other pollens (92%; n=289), whereas only 64.2% were positive in the group of multiple pollen sensitised subjects (MPS; n=204). Differences were recorded in geographical distribution of rPar j 2 reactive subjects. A relatively small area with statistically significant lower prevalence rPar j 2 IgE positive subjects was identified when native subjects were considered. Correlating IgE results, PjE vs rPar j 2, the best result was achieved in the mono-sensitised and oligo-sensitised cohort again if compared to the MPS patient subset. When results from mono-sensitised subjects were plotted, three further subgroups were identified. A first group with almost overlapping values for both PjE and rPar j 2, a second with PjE values greater than rPar j 2 values, and a third group with the opposite finding. Representative sera from these three subgroups have been tested for both rPar j 1 and 2. All reacted with both. rPar j 2 can be assumed as a specific marker for exposure and sensitisation to Parietaria pollen. rPar j 2 evaluation allows both a more precise geographical mapping and for identification of Parietaria sensitised subjects within MPS subset. Further studies are needed to define whether a single molecule (rPar j 2) is suitable for both diagnosis and future molecule-based immunotherapy.

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[47] - Bonura A, Artale A, Marino M, Amoroso S, Marcucci F, Geraci D, et al. Cross-reactivity between Parietaria species using the major rPar j 1 and rPar j 2 allergens. Allergy Asthma Proc 2006;27:378-382

Parietaria pollen is one of the most important outdoor allergenic sources in all the Mediterranean countries, with a large number of subjects showing a positive skin prick test to both P. judaica and P. officinalis pollen extracts. A cross-reactivity between the two species has been already reported although few data are known at the molecular level. Twenty-five consecutive patients with Parietaria pollen allergy were selected on the basis of their clinical history. Skin prick test to P. judaica and officinalis extracts was performed. In vitro IgE measurement to both allergenic sources was performed by using a quantitative assay. ELISA inhibition experiments were made by using the rParj1 and rParj2 allergens. All the patients showed a positive skin prick test to both Parietaria species. Quantitative IgE measurement showed similar antibody concentration for P. judaica and P. officinalis extracts. ELISA inhibition experiments demonstrated that the cross-reactivity between the two species was due to the presence of the Parj1 and Parj2 allergens in the extracts with a high conserved IgE epitopes content. We conclude that P. judaica and P. officinalis pollens contain highly cross-reactive species-specific major allergens useful for the diagnosis and therapy of both allergenic sources.

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[48] - Asero R, Mistrello G, Roncarolo D, Amato S. Parietaria Profilin Shows Only Limited Cross-Reactivity with Birch and Grass Profilins. Int Arch Allergy Immunol 2004;133:121-124

BACKGROUND: In patients showing multiple reactivity to seasonal airborne allergens, a skin prick test (SPT) with Parietaria pollen scores frequently negative, suggesting that profilin from Parietaria might not share IgE-binding epitopes with profilin from botanically unrelated airborne allergens. This study investigated the immunologic cross-reactivity between profilins from Parietaria, grass and birch pollen . METHODS: 36 patients hypersensitive to birch profilin, Bet v 2, underwent SPT with Parietaria, and IgE to both whole Parietaria pollen and Phleum profilin were sought in their sera by ELISA. In ELISA inhibition studies, IgE reactivity to Phleum profilin was measured before and after absorption of sera from Parietaria reactors with both whole Parietaria pollen extract, and Par j 1/Par j 2 fraction. Further, their IgE reactivity to whole Parietaria pollen was measured before and after absorption with the Par j 1/Par j 2 fraction . RESULTS: All sera showed IgE reactivity to Phleum profilin. Only 14/36 (39%) patients had a positive SPT to Parietaria and 17/36 (47%) showed IgE to Parietaria. Absorption of sera from Parietaria reactors with whole Parietaria extract caused a marked reduction in IgE reactivity to grass profilin; in contrast, absorption of sera with the Par j 1/Par j 2 fraction did not exert any inhibitory effect on IgE reactivity to grass profilin. Absorption of sera with the Par j 1/Par j 2 fraction markedly reduced IgE reactivity to whole Parietaria extract in 8/9 cases . CONCLUSION: Less than 50% of patients sensitized to birch and grass profilin cross-react to Parietaria profilin. In most cases, cross-reactors are hypersensitive to major, specific Parietaria allergens as well. This findings may be of practical relevance when the prescription of specific immunotherapy is considered.

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[49] - Duro G, Colombo P, Costa MA, Izzo V, Porcasi R, Di Fiore R, et al. cDNA cloning, sequence analysis and allergological characterization of Par j 2.0101, a new major allergen of the Parietaria judaica pollen. FEBS Lett 1996;399:295-298

A clone (P2) coding for an allergen of Parietaria judaica (Pj) pollen has been isolated and sequenced from a cDNA library in lambda ZAP using a pool of 23 sera from Pj-allergic patients. The clone contained an insert of 622 nucleotides with an open reading frame of 133 amino acids (aa) and a putative signal peptide of 31 aa giving a deduced mature processed protein of 102 aa with a molecular mass of 11344 Da. The expressed recombinant protein, named rPar j 2.0101, was a major allergen since it reacted with IgE of 82% (23/28) of the sera of Pj- allergic subjects analyzed. It was shown to be a new allergen since (i) the amino acid sequence homology with the already reported recombinant allergen Par j 1.0101 was 45% and (ii) there was no cross-inhibition between rPar j 2.0101 and rPar j 1.0101. In addition, rPar j 2.0101 inhibited 35% of the specific IgE for 10-14 kDa native allergens and preincubation of sera from Pj-allergic patients with both rPar j 2.0101 and rPar j 1.0101 fully abolished the IgE recognition of the 10-14 kDa native allergen region, suggesting that these two allergens contributed to the region

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[50] - Stumvoll S, Westritschnig K, Lidholm J, Spitzauer S, Colombo P, Duro G, et al. Identification of cross-reactive and genuine Parietaria judaica pollen allergens. J Allergy Clin Immunol 2003;111:974-979

BACKGROUND: The weed Parietaria judaica is one of the most important pollen allergen sources in the Mediterranean area. OBJECTIVE: We sought to identify P judaica pollen allergen, which might be used to serologically distinguish genuine Parietaria sensitization and cross-reactivity to allergens from other weed species (eg, mugwort and ragweed). METHODS: The allergen profile of P judaica IgE-reactive sera from weed pollen-sensitized allergic individuals from the Mediterranean region (n = 36) with high Parietaria pollen exposure and from weed pollen-allergic patients with little or no Parietaria exposure (Austria, n = 42; Scandinavia, n = 8; United States, n = 19) was established by CAP FEIA measurements and by IgE immunoblot inhibition experiments with recombinant allergens. RESULTS: The majority (83%) of the Mediterranean weed pollen-allergic patients mounted high IgE antibody levels (mean specific IgE, 20.89 kUA/L) against recombinant (r) Par j 2, whereas only 7% of the non-Mediterranean weed-allergic patients showed low IgE reactivity to rPar j 2 (mean specific IgE, 1.03 kUA/L). The cytoskeletal protein profilin and a 2-EF-hand calcium-binding allergen were identified as cross-reactive Parietaria allergens, which were recognized preferentially by Parietaria -positive, non-Mediterranean weed pollen-allergic patients. CONCLUSION: rPar j 2 might be used as a diagnostic marker allergen to identify weed pollen-allergic patients who are genuinely sensitized against Parietaria pollen and thus would be particularly suited for specific immunotherapy with Parietaria pollen extract.

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[51] - Amoresano A, Pucci P, Duro G, Colombo P, Costa MA, Izzo V, et al. Assignment of disulphide bridges in Par j 2.0101, a major allergen of Parietaria judaica pollen. Biol Chem 2003;384:1165-1172

Par j 2.0101, a major allergen of the Parietaria judaica pollen, was expressed in E. coli, purified to homogeneity and fully characterised both at the structural and the functional level. The recombinant rPar j 2.0101 protein showed an allergenic activity in histamine release, skin prick tests and capacity to bind IgE, almost identical to that of the native allergens purified from aqueous pollen extract. The complete pattern of S-S bridges of rPar j 2.0101 was determined by enzymatic digestion with endoproteinase Lys-C followed by mass spectrometric analysis of the resulting peptide mixtures. The eight cysteines occurring in the allergenic protein were found to be paired into the following four disulphides: Cys35-Cys83, Cys45-Cys60, Cys61-Cys106 and Cys81-Cys121. This structural information probes Par j 2.0101 to attain a 3-D fold consistent with that of the non-specific lipid transfer protein (ns-LTP) family and it represents an effective molecular basis to develop modified antigens by selective site-directed mutagenesis for immunotherapy.

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[52] - Gonzalez-Rioja R, Ferrer A, Arilla MC, Ibarrola I, Viguera AR, Andreu C, et al. Diagnosis of Parietaria judaica pollen allergy using natural and recombinant Par j 1 and Par j 2 allergens. Clin Exp Allergy 2007;37:243-250

Background Parietaria judaica pollen is one of the main causes of allergic diseases in the Mediterranean area and contains two major allergens, called Par j 1 and Par j 2. Objective To evaluate the diagnostic potential of natural and recombinant forms of Par j 1 and Par j 2 in comparison with standardized P. judaica pollen extract. Methods Thirty patients allergic to P. judaica pollen and 15 control patients were investigated. Skin prick tests and determination of specific IgE levels were performed with commercial P. judaica extract, natural Par j 1 and Par j 2, and recombinant forms of both allergens expressed in P. pastoris. Results The whole group of patients with allergy to P. judaica had a positive skin test reaction to purified nPar j 1-Par j 2 and rPar j 2 at 5 mug/mL, and no false-positive reactions were detected. Natural and recombinant Par j 1 and Par j 2 showed no significantly different responses in skin tests compared with P. judaica extract. A high correlation was found between the serum-specific IgE levels to P. judaica extract vs. natural (R=0.996; P<0.001) and recombinant allergens (R=0.887 and 0.982 for rPar j 1 and rPar j 2, respectively; P<0.001). rPar j 2 displayed a 100% sensitivity and specificity among P. judaica-allergic patients. Conclusions In vivo and in vitro diagnosis of P. judaica pollen allergy could be simplified using rPar j 2. This protein showed comparable IgE response and skin prick reactivity with those produced by P. judaica pollen extract.

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[53] - Asturias JA, Ibarrola I, Eseverri JL, Arilla MC, Gonzalez-Rioja R, Martinez A. PCR-based cloning and immunological characterization of Parietaria judaica pollen profilin. J Investig Allergol Clin Immunol 2004;14:43-48

Profilin has been described as an allergen present in pollen of trees, grasses and weeds. Since Parietaria judaica profilin has a molecular mass similar to other Parietaria allergens (Par j 1 and Par j 2) in the 14-10 kDa range, it is difficult to assess the prevalence of profilin by immunoblotting or to obtain sufficient amounts of purified native profilin for investigation and diagnosis. The aim of this study was to identify P. judaica profilin by PCR-based cDNA cloning and to elucidate its allergenic characteristics. Two cDNA clones encoding P. judaica pollen profilin were isolated by polymerase chain reaction (PCR) amplification using degenerate primers. Sequencing of both clones (Par j 3.0101 and Par j 3.0102) demonstrated a high amino acid sequence homology. Immunodetection of P. judaica pollen after isoelectrofocusing and incubation with rabbit antiserum against profilin indicated the existence of at least 2 isoforms. Expression in Escherichia coli BL21 (DE3) was carried out using a vector based in the T7 expression system, and the recombinant allergen was isolated by affinity chromatography on poly-(L-proline)-Sepharose. Cross-reactivity has been found between recombinant P. judaica pollen profilin and profilins from other botanical unrelated plants.

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[54] - Bonura A, Gulino L, Trapani A, Di Felice G, Tinghino R, Amoroso S, et al. Isolation, expression and immunological characterization of a calcium-binding protein from Parietaria pollen. Mol Immunol 2008;45:2465-2473

The diagnosis and therapy of allergic disorders are usually performed with crude extracts which are a heterogeneous mixture of proteins with different allergenic potency. The knowledge of the allergenic composition is a key step for diagnostic and therapeutic options. Parietaria judaica pollen represents one of the main sources of allergens in the Mediterranean area and its major allergens have already been identified (Par j 1 and Par j 2). In addition, inhibition studies performed using a calcium-binding protein (CBP) from grass pollen (Phl p 7) showed the presence of a homologue of this cross-reactive allergen in the Parietaria extract. Screening of a cDNA library allowed us to isolate a 480bp cDNA containing the information for an 87 AA long protein with high level of homology to calcium-binding proteins from other allergenic sources. It was expressed as a recombinant allergen in Escherichia coli and purified by affinity chromatography. Its expression allowed us to study the prevalence of this allergen in a population of allergic patients in southern Europe. Immunoblotting and inhibition studies showed that this allergen shares a pattern of IgE epitopes in common with other 2-EF-hand calcium-binding proteins from botanically non-related species. The immunological properties of the Pj CBP were investigated by CD63 activation assay and CFDA-SE staining. In conclusion, DNA recombinant technology allowed the isolation, expression and immunological characterization of a cross-reactive calcium-binding protein allergen from Parietaria judaica pollen.

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[55] - Stumvoll S, Westritschnig K, Lidholm J, Spitzauer S, Colombo P, Duro G, et al. Identification of cross-reactive and genuine Parietaria judaica pollen allergens. J Allergy Clin Immunol 2003;111:974-979

BACKGROUND: The weed Parietaria judaica is one of the most important pollen allergen sources in the Mediterranean area. OBJECTIVE: We sought to identify P judaica pollen allergen, which might be used to serologically distinguish genuine Parietaria sensitization and cross-reactivity to allergens from other weed species (eg, mugwort and ragweed). METHODS: The allergen profile of P judaica IgE-reactive sera from weed pollen-sensitized allergic individuals from the Mediterranean region (n = 36) with high Parietaria pollen exposure and from weed pollen-allergic patients with little or no Parietaria exposure (Austria, n = 42; Scandinavia, n = 8; United States, n = 19) was established by CAP FEIA measurements and by IgE immunoblot inhibition experiments with recombinant allergens. RESULTS: The majority (83%) of the Mediterranean weed pollen-allergic patients mounted high IgE antibody levels (mean specific IgE, 20.89 kUA/L) against recombinant (r) Par j 2, whereas only 7% of the non-Mediterranean weed-allergic patients showed low IgE reactivity to rPar j 2 (mean specific IgE, 1.03 kUA/L). The cytoskeletal protein profilin and a 2-EF-hand calcium-binding allergen were identified as cross-reactive Parietaria allergens, which were recognized preferentially by Parietaria -positive, non-Mediterranean weed pollen-allergic patients. CONCLUSION: rPar j 2 might be used as a diagnostic marker allergen to identify weed pollen-allergic patients who are genuinely sensitized against Parietaria pollen and thus would be particularly suited for specific immunotherapy with Parietaria pollen extract.

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[56] - Rossi RE, Monasterolo G, Coco G, Operti D. Possible Relationship between Systemic Side Effects and Sensitization to rPar j 2 in Allergic Patients Submitted to an Ultra-Rush (20 min) Sublingual Immunotherapy and Selected by Component Resolved Diagnosis. Int Arch Allergy Immunol 2005;138:105-110

BACKGROUND: The pollen of Parietaria spp., a weed of the Urticaceae family, is a major cause of respiratory allergy in the Mediterranean area, where the most common species are Parietaria judaica and Parietaria officinalis. In this study, we evaluated the specific serum IgE-binding profiles to individual P. judaica pollen recombinant major allergen, and Phleum pratense cytoskeletal profilin and a 2-EF-hand calcium-binding allergen homologous to cross-reactive Parietaria pollen allergens, in patients allergic to pollen with positive skin test towards Parietaria spp. extract . METHODS: The present observation included 220 patients from the province of Cuneo, north-west Italy, all suffering from rhino-conjunctivitis and/or asthma selected on the basis of skin test positive to P. judaica extract. The sera were evaluated for specific IgE reactivity to P. judaica pollen major recombinant(r) allergen Par j 2, Phleum pratense pollen allergens rPhl p 7 (2-EF-hand calcium binding protein) and rPhl p 12 (profilin), both identified as cross-reactive Parietaria spp. allergens, using Pharmacia CAP System. Out of 220 patients, 37 patients with IgE reactivity to rPar j 2 and 105 patients sensitized to at least one timothy pollen major allergen (i. e. rPhl p 1, rPhl p 2, natural Phl p 4 and rPhl p 6) were submitted to an ultra-rush protocol of sublingual immunotherapy (SLIT). The occurrence of adverse reactions were evaluated in both groups . RESULTS: All 220 patients with pollinosis and positive in vivo skin prick tests had in vitro positive CAP results to P. judaica natural extract. On the contrary, in these patients the prevalence of Par j 2-specific IgE was only 33.2% (73/220). In fact, 116/220 (52.7%) patients with serum specific IgE to crude Parietaria pollen extract had specific IgE to Phl p 12, 18/220 (8.1%) subjects with specific IgE to rPhl p 12 also exhibited specific IgE to Phl p 7 and 26/220 (11.8%) subjects had specific IgE against rPhl p 7. Particularly, geometric mean (25th-75th percentile) of specific IgE to rPar j 2 were as follows: 2.87 kUA/l (1.005-7.465). Out of 73 patients with specific IgE to rPar j 2, 7 subjects (9.6%) had also specific IgE to rPhl p 7, 12 (16.4%) had specific IgE to rPhl p 12 and 4 (4.1%) patients had specific IgE to both recombinant allergens. Of 37 patients under an ultra-rush protocol of SLIT, 3 subjects (8.1%) experienced generalized urticaria, and 1 of them also had diarrhea 3 h after the last dose of Parietaria judaica extract oral-vaccine administration. On the contrary, no systemic reactions were observed in 105 patients after Phleum pratense extract oral intake after a similar ultra-rush SLIT protocol (p = 0.0046) . CONCLUSIONS: In the light of present findings, allergen extract-based diagnosis, in vivo and in vitro, cannot discriminate allergic patients that are genuinely sensitized to Parietaria spp. major allergens or to other major allergens to which current immunotherapeutic allergy extracts are standardized. Therefore, in vitro component resolved diagnosis is the unique tool to define the disease eliciting molecule(s). Finally, during sublingual immunotherapy, not only the dose of allergen, but also the biochemical characteristic of the major allergen administered may be an important factor in determining possible systemic reactions.

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[57] - Berrens L, Maranon F. IgE-binding trypsin inhibitors in plant pollen extracts. Experientia 1995;51:953-955

In an attempt to access the possible role of protease-antiprotease mechanisms of non-immune defence in pollinosis, only low levels of trypsin-, kallikrein- or plasmin-like proteinases could be detected in aqueous pollen extracts. In contrast, several pollen species displayed appreciable trypsin inhibitory activity, e.g. Parietaria, Olea, Ambrosia, Rumex, Chenopodium, Holcus and Poa spp. These proteins of the serpin family of anti-proteinases were found to bind specific IgE-antibodies from the serum of hay fever patients. As examples, the IgE-binding trypsin inhibitors from the pollen of Parietaria judaica and Ambrosia elatior were purified and characterized as acidic proteins with pI 4.2 and a molecular weight of 20-24 kDa

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[58] - Tinghino R, Barletta B, Palumbo S, Afferni C, Iacovacci P, Mari A, et al. Molecular characterization of a cross-reactive Juniperus oxycedrus pollen allergen, Jun o 2: a novel calcium-binding allergen. J Allergy Clin Immunol 1998;101:772-777

BACKGROUND: Species belonging to the Cupressaceae family are a relevant source of allergens that are present in a wide number of countries. OBJECTIVE: We sought to identify, purify, and characterize recombinant allergens from Juniperus oxycedrus, a species belonging to the Cupressaceae family. METHODS: Double-stranded cDNA was synthesized from mRNA and cloned into the lambda-ZAP expression vector. IgE screening of the library was performed with a pool of sera from subjects allergic to Cupressaceae. A recombinant 6xHis-tagged Juniperus oxycedrus allergen, Jun o 2, was expressed in Escherichia coli and purified by Ni2+ affinity chromatography. It was studied further by immunoblotting inhibition with pollen extracts from other Cupressaceae, Oleaceae, Urticaceae, and Graminaceae. The role of protein-bound calcium on the allergen's IgE-binding capacity was tested in a plaque assay in the presence or absence of EGTA. RESULTS: A cDNA coding for a newly identified Juniperus oxycedrus pollen allergen, rJun o 2, was isolated. The deduced amino acid sequence contained four typical Ca2+ binding sites and showed a significant sequence similarity to calmodulins. Depletion of Ca2+ in the plaque assay led to a loss of IgE-binding capacity of rJun o 2. Immunoblotting inhibition revealed that J. oxycedrus, J. ashei, Cupressus arizonica, C. sempervirens, Parietaria judaica, Olea europaea, and Lolium perenne pollen extracts were able to inhibit IgE binding to blotted rJun o 2 at different concentrations. CONCLUSION: rJun o 2 contains IgE-binding epitopes shared by taxonomically unrelated species, and therefore it can be regarded as a new panallergen. These findings could contribute to an explanation for the phenomenon of multiple positive test results in polysensitized patients and the potential symptom-eliciting role of allergenic sources previously not encountered.

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[59] - Tinghino R, Twardosz A, Barletta B, Puggioni EMR, Iacovacci P, Afferni C, et al. Molecular, structural, and immunologic relationships between different families of recombinant calcium-binding pollen allergens. J Allergy Clin Immunol 2002;109:314-320

Background: Calcium-binding plant allergens can be grouped in different families according to the number of calciumbinding domains (EF hands). Objective: We sought to identify pollens containing crossreactive calcium-binding allergens and to investigate structural and immunologic similarities of members belonging to different families of calcium-binding allergens. Methods: By means of multiple sequence alignment and molecular modeling, we searched for structural similarities among pollen allergens with 2 (Phl p 7, timothy grass; Aln g 4, alder), 3 (Bet v 3, birch) and 4 EF hands (Jun o 4, prickly juniper). Purified recombinant Aln g 4 and Jun o 4 were used to determine the prevalence of IgE recognition in 210 patients sensitized to different pollens and to search, by means of ELISA competition, for the presence of cross-reactive epitopes in pollens from 16 unrelated plant species. IgE cross-reactivity among the allergen families was studied with purified rPhl p 7, rAln g 4, rBet v 3, and rJun o 4 and 2 synthetic peptides comprising the N-terminal and C-terminal EF hands of Phl p 7 by means of ELISA competition. Results: Structural similarities were found by using molecular modeling among the allergens with 2, 3, and 4 EF hands. Pollens from 16 unrelated plants contained Aln g 4- and Jun o 4-related epitopes. Twenty-two percent of the patients with multiple pollen sensitization reacted to at least one of the calcium-binding allergens. A hierarchy of IgE cross-reactivity (rPhl p7 > rAln g 4 > rJun o 4 > rBet v 3) could be established that identified rPhl p 7 as the EF-hand allergen containing most IgE epitopes in the population studied. Conclusion: The demonstration that members of different families of calcium-binding plant allergens share similarities suggests that it may be possible to use representative molecules for the diagnosis and therapy of allergies to EF-hand allergens.

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[60] - Tinghino R, Hayek B, Barletta B, Puggioni E, Iacovacci P, Butteroni C, et al. Ca-binding allergens: cross-reactivity between molecules with 2 (rAln g4) and 4 (rJun o2) EF-hand domains. J Allergy Clin Immunol 2000;105(1 part 2):S332

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[61] - Tinghino R, Twardosz A, Barletta B, Puggioni EMR, Iacovacci P, Afferni C, et al. Molecular, structural, and immunologic relationships between different families of recombinant calcium-binding pollen allergens. J Allergy Clin Immunol 2002;109:314-320

Background: Calcium-binding plant allergens can be grouped in different families according to the number of calciumbinding domains (EF hands). Objective: We sought to identify pollens containing crossreactive calcium-binding allergens and to investigate structural and immunologic similarities of members belonging to different families of calcium-binding allergens. Methods: By means of multiple sequence alignment and molecular modeling, we searched for structural similarities among pollen allergens with 2 (Phl p 7, timothy grass; Aln g 4, alder), 3 (Bet v 3, birch) and 4 EF hands (Jun o 4, prickly juniper). Purified recombinant Aln g 4 and Jun o 4 were used to determine the prevalence of IgE recognition in 210 patients sensitized to different pollens and to search, by means of ELISA competition, for the presence of cross-reactive epitopes in pollens from 16 unrelated plant species. IgE cross-reactivity among the allergen families was studied with purified rPhl p 7, rAln g 4, rBet v 3, and rJun o 4 and 2 synthetic peptides comprising the N-terminal and C-terminal EF hands of Phl p 7 by means of ELISA competition. Results: Structural similarities were found by using molecular modeling among the allergens with 2, 3, and 4 EF hands. Pollens from 16 unrelated plants contained Aln g 4- and Jun o 4-related epitopes. Twenty-two percent of the patients with multiple pollen sensitization reacted to at least one of the calcium-binding allergens. A hierarchy of IgE cross-reactivity (rPhl p7 > rAln g 4 > rJun o 4 > rBet v 3) could be established that identified rPhl p 7 as the EF-hand allergen containing most IgE epitopes in the population studied. Conclusion: The demonstration that members of different families of calcium-binding plant allergens share similarities suggests that it may be possible to use representative molecules for the diagnosis and therapy of allergies to EF-hand allergens.

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[62] - Bonura A, Gulino L, Trapani A, Di Felice G, Tinghino R, Amoroso S, et al. Isolation, expression and immunological characterization of a calcium-binding protein from Parietaria pollen. Mol Immunol 2008;45:2465-2473

The diagnosis and therapy of allergic disorders are usually performed with crude extracts which are a heterogeneous mixture of proteins with different allergenic potency. The knowledge of the allergenic composition is a key step for diagnostic and therapeutic options. Parietaria judaica pollen represents one of the main sources of allergens in the Mediterranean area and its major allergens have already been identified (Par j 1 and Par j 2). In addition, inhibition studies performed using a calcium-binding protein (CBP) from grass pollen (Phl p 7) showed the presence of a homologue of this cross-reactive allergen in the Parietaria extract. Screening of a cDNA library allowed us to isolate a 480bp cDNA containing the information for an 87 AA long protein with high level of homology to calcium-binding proteins from other allergenic sources. It was expressed as a recombinant allergen in Escherichia coli and purified by affinity chromatography. Its expression allowed us to study the prevalence of this allergen in a population of allergic patients in southern Europe. Immunoblotting and inhibition studies showed that this allergen shares a pattern of IgE epitopes in common with other 2-EF-hand calcium-binding proteins from botanically non-related species. The immunological properties of the Pj CBP were investigated by CD63 activation assay and CFDA-SE staining. In conclusion, DNA recombinant technology allowed the isolation, expression and immunological characterization of a cross-reactive calcium-binding protein allergen from Parietaria judaica pollen.

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[63] - Bonura A, Artale A, Marino M, Amoroso S, Marcucci F, Geraci D, et al. Cross-reactivity between Parietaria species using the major rPar j 1 and rPar j 2 allergens. Allergy Asthma Proc 2006;27:378-382

Parietaria pollen is one of the most important outdoor allergenic sources in all the Mediterranean countries, with a large number of subjects showing a positive skin prick test to both P. judaica and P. officinalis pollen extracts. A cross-reactivity between the two species has been already reported although few data are known at the molecular level. Twenty-five consecutive patients with Parietaria pollen allergy were selected on the basis of their clinical history. Skin prick test to P. judaica and officinalis extracts was performed. In vitro IgE measurement to both allergenic sources was performed by using a quantitative assay. ELISA inhibition experiments were made by using the rParj1 and rParj2 allergens. All the patients showed a positive skin prick test to both Parietaria species. Quantitative IgE measurement showed similar antibody concentration for P. judaica and P. officinalis extracts. ELISA inhibition experiments demonstrated that the cross-reactivity between the two species was due to the presence of the Parj1 and Parj2 allergens in the extracts with a high conserved IgE epitopes content. We conclude that P. judaica and P. officinalis pollens contain highly cross-reactive species-specific major allergens useful for the diagnosis and therapy of both allergenic sources.

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[64] - Bousquet J, Hewitt B, Guerin B, Dhivert H, Michel FB. Allergy in the Mediterranean area. II: Cross-allergenicity among Urticaceae pollens (Parietaria and Urtica). Clin Allergy 1986;16:57-64

Pollens of related species usually share common antigens and are cross-reacting but this is not always the case. The cross-allergenicity of two species of Urticaceae pollens (Parietaria and Urtica, nettle) was sought by means of RAST-inhibition and isoelectric focusing. A preliminary experiment in forty-two patients investigated by means of skin tests and RAST showed that most patients were either allergic to Parietaria or nettle and that both reactivities were uncommonly associated. The results of RAST-inhibition confirmed the total absence of cross-reactivity between these two related species. IEF patterns indicated that most proteins are in the acidic range of pH and did not reveal large discrepancies. Parietaria and Urtica pollens, although belonging to the same family and having close microscopic patterns, have very different allergenic activities. This study demonstrates that cross-allergenicity cannot be predicted.

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[65] - Vega-Maray AM, Fernandez-Gonzalez D, Valencia-Barrera R, Suarez-Cervera M. Allergenic proteins in Urtica dioica, a member of the Urticaceae allergenic family. Ann Allergy Asthma Immunol 2006;97:343-349

BACKGROUND: Allergy to the pollen of flowering plant species significantly affects the health of people in many parts of the world. Pollens of related genera usually share common antigens and are often, but not always, cross-reactive. Several studies have shown that Parietaria pollen is one of the most common causes of pollinosis in the Mediterranean area, whereas Urtica has no allergenic significance. OBJECTIVES: To report on the localization of Parietaria judaica major allergen in Urtica dioica pollen grains and on the detection of allergenic proteins in U. dioica pollen grains during the hydration-activation process. METHODS: A combination of transmission electron microscopy and immunocytochemical methods was used to locate allergenic proteins in U. dioica pollen grains after different periods of hydration-activation using the anti-Par j 1 (4.1.3.) monoclonal antibody and serum samples from allergic patients. RESULTS: No significant labeling was noted for Parj 1 allergen after 10, 15, and 20 minutes in the walls and cytoplasm. Slight labeling was observed for allergic proteins in the walls of U. dioica after 10 minutes of hydration, and no significant labeling was found after 15 and 20 minutes of hydration. CONCLUSIONS: Immunocytochemical methods confirmed the absence of cross-reactivity between 2 related genera, Parietaria and Urtica, and the lowest allergenic potential of U. dioica.

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[66] - Miura N. Ramie (Boehmeria nivea) pollen-induced bronchial asthma and allergenic cross-reactivity of ramie and Parietaria. Arerugi 1993;42:649-655

Ramie (Boehmeria nivea), a plant of the Urticaceae family, is widely distributed in the Nagasaki area, and has been established to be a cause of astham. The rate of positive reactions to ramie in intradermal tests was 11.7% among adult asthmatic patients in the Nagasaki area. In this study, 10 patients were positive in provocation tests using ramie pollen Ramie pollen-specific IgE antibodies were measured by ELISA, with the positive provocation test group showing higher O.D. values than the positive intradermal test group (p < 0.05). Ramie is of the same family as Parietaria, an important allergen in Europe. The cross-reactivity of ramie and Parietaria was examined by an ELISA inhibition test using P officinalis and P. judaica (Pj10), but no cross-reactivity was found, suggesting that ramie may be a new independent allergen. As ramie is widely distributed throughout Japan and South-east Asia, further study is needed to determine whether it is an important allergen of the Urticaceae family in this region, as is Parietaria in Europe, and Urtica in America.

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[67] - Holgate ST, Jackson L, Watson HK, Ganderton MA. Sensitivity to Parietaria pollen in the Southampton area as determined by skin-prick and RAST tests. Clin Allergy 1988;18:549-556

The pollen of Parietaria species is a well-recognized and important inhalant allergen in the Mediterranean area. Parietaria judaica (Pellitory-of-the-Wall) is native to the U.K., flowering from June to September, but is not usually considered to be of any clinical importance by U.K. allergists. We skin tested 62 patients with a clinical history of summer seasonal respiratory symptoms and a control group of 11 patients with perennial respiratory symptoms only. Each was skin tested in duplicate with extracts of grass pollen, birch pollen, Parietaria pollen, Dermatophagoides pteronyssinus, Cladosporium, Alternaria, nettle pollen and negative and positive controls, and serum samples were collected for RAST assays for Parietaria and nettle. Eight of the 62 patients in the main group showed skin reactivity to Parietaria. Five of these eight had never visited the Mediterranean area and therefore it is possible that sensitization occurred in the U.K. Thirteen of the 62 patients were skin reactive to nettle but there was no correlation between skin reactivity to Parietaria and nettle. This supports a recent report that, despite their close botanical relationship, no antigenic cross-reactivity exists between the two species. No correlation was seen between skin reactivity and serum RAST activity to Parietaria or nettle. It is not known whether exposure to Parietaria pollen contributes to the seasonal symptoms of the patients found to be skin reactive. None of the 11 patients in the control group was skin reactive to Parietaria.

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[68] - Pham NH, Baldo BA. Allergenic relationship between taxonomically diverse pollens. Clin Exp Allergy 1995;25:599-606

Skin tests and tests for IgE antibodies show that subjects are usually sensitive to a number of different pollens, frequently from taxonomically diverse species which are assumed to be allergenically non-crossreactive. This suggests that the presence of IgE antibody-reactivity to an individual pollen may not necessarily have resulted from contact with that pollen or even with a taxonomically closely related species. OBJECTIVE: Since this has important consequences for allergen avoidance and desensitization of patients, we attempted to define allergenic relationships between diverse pollen species. METHODS: Sera from subjects were examined in direct IgE antibody binding experiments and by quantitative inhibition, protein blotting and adsorption and elution studies. RESULTS: Sera from subjects diagnosed as allergic to white cypress pine, Italian cypress, ryegrass or birch pollen were shown to have IgE antibodies that reacted with pollens from these four species and from cocksfoot, couch grass, lamb's quarter, wall pellitory, olive, plantain and ragweed. These reactions were confirmed in protein blotting and adsorption and elution studies where numerous IgE-binding bands were detected in all 11 different pollen extracts with sera from each of the different allergic categories. Further evidence of allergenic (i.e. IgE-binding crossreactivity between the different pollens was provided by inhibition studies in which clear-cut inhibitions of IgE binding to the different pollen allergen discs were obtained with comparable amounts of the different pollen extracts. CONCLUSION: We conclude that the presence of pollen reactive IgE antibodies may not necessarily be a true reflection of sensitizing pollen species.

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[69] - Stumvoll S, Westritschnig K, Lidholm J, Spitzauer S, Colombo P, Duro G, et al. Identification of cross-reactive and genuine Parietaria judaica pollen allergens. J Allergy Clin Immunol 2003;111:974-979

BACKGROUND: The weed Parietaria judaica is one of the most important pollen allergen sources in the Mediterranean area. OBJECTIVE: We sought to identify P judaica pollen allergen, which might be used to serologically distinguish genuine Parietaria sensitization and cross-reactivity to allergens from other weed species (eg, mugwort and ragweed). METHODS: The allergen profile of P judaica IgE-reactive sera from weed pollen-sensitized allergic individuals from the Mediterranean region (n = 36) with high Parietaria pollen exposure and from weed pollen-allergic patients with little or no Parietaria exposure (Austria, n = 42; Scandinavia, n = 8; United States, n = 19) was established by CAP FEIA measurements and by IgE immunoblot inhibition experiments with recombinant allergens. RESULTS: The majority (83%) of the Mediterranean weed pollen-allergic patients mounted high IgE antibody levels (mean specific IgE, 20.89 kUA/L) against recombinant (r) Par j 2, whereas only 7% of the non-Mediterranean weed-allergic patients showed low IgE reactivity to rPar j 2 (mean specific IgE, 1.03 kUA/L). The cytoskeletal protein profilin and a 2-EF-hand calcium-binding allergen were identified as cross-reactive Parietaria allergens, which were recognized preferentially by Parietaria -positive, non-Mediterranean weed pollen-allergic patients. CONCLUSION: rPar j 2 might be used as a diagnostic marker allergen to identify weed pollen-allergic patients who are genuinely sensitized against Parietaria pollen and thus would be particularly suited for specific immunotherapy with Parietaria pollen extract.

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[70] - Asero R, Mistrello G, Roncarolo D, Amato S. Parietaria Profilin Shows Only Limited Cross-Reactivity with Birch and Grass Profilins. Int Arch Allergy Immunol 2004;133:121-124

BACKGROUND: In patients showing multiple reactivity to seasonal airborne allergens, a skin prick test (SPT) with Parietaria pollen scores frequently negative, suggesting that profilin from Parietaria might not share IgE-binding epitopes with profilin from botanically unrelated airborne allergens. This study investigated the immunologic cross-reactivity between profilins from Parietaria, grass and birch pollen . METHODS: 36 patients hypersensitive to birch profilin, Bet v 2, underwent SPT with Parietaria, and IgE to both whole Parietaria pollen and Phleum profilin were sought in their sera by ELISA. In ELISA inhibition studies, IgE reactivity to Phleum profilin was measured before and after absorption of sera from Parietaria reactors with both whole Parietaria pollen extract, and Par j 1/Par j 2 fraction. Further, their IgE reactivity to whole Parietaria pollen was measured before and after absorption with the Par j 1/Par j 2 fraction . RESULTS: All sera showed IgE reactivity to Phleum profilin. Only 14/36 (39%) patients had a positive SPT to Parietaria and 17/36 (47%) showed IgE to Parietaria. Absorption of sera from Parietaria reactors with whole Parietaria extract caused a marked reduction in IgE reactivity to grass profilin; in contrast, absorption of sera with the Par j 1/Par j 2 fraction did not exert any inhibitory effect on IgE reactivity to grass profilin. Absorption of sera with the Par j 1/Par j 2 fraction markedly reduced IgE reactivity to whole Parietaria extract in 8/9 cases . CONCLUSION: Less than 50% of patients sensitized to birch and grass profilin cross-react to Parietaria profilin. In most cases, cross-reactors are hypersensitive to major, specific Parietaria allergens as well. This findings may be of practical relevance when the prescription of specific immunotherapy is considered.

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[71] - Pham NH, Baldo BA. Allergenic relationship between taxonomically diverse pollens. Clin Exp Allergy 1995;25:599-606

Skin tests and tests for IgE antibodies show that subjects are usually sensitive to a number of different pollens, frequently from taxonomically diverse species which are assumed to be allergenically non-crossreactive. This suggests that the presence of IgE antibody-reactivity to an individual pollen may not necessarily have resulted from contact with that pollen or even with a taxonomically closely related species. OBJECTIVE: Since this has important consequences for allergen avoidance and desensitization of patients, we attempted to define allergenic relationships between diverse pollen species. METHODS: Sera from subjects were examined in direct IgE antibody binding experiments and by quantitative inhibition, protein blotting and adsorption and elution studies. RESULTS: Sera from subjects diagnosed as allergic to white cypress pine, Italian cypress, ryegrass or birch pollen were shown to have IgE antibodies that reacted with pollens from these four species and from cocksfoot, couch grass, lamb's quarter, wall pellitory, olive, plantain and ragweed. These reactions were confirmed in protein blotting and adsorption and elution studies where numerous IgE-binding bands were detected in all 11 different pollen extracts with sera from each of the different allergic categories. Further evidence of allergenic (i.e. IgE-binding crossreactivity between the different pollens was provided by inhibition studies in which clear-cut inhibitions of IgE binding to the different pollen allergen discs were obtained with comparable amounts of the different pollen extracts. CONCLUSION: We conclude that the presence of pollen reactive IgE antibodies may not necessarily be a true reflection of sensitizing pollen species.

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[72] - Tinghino R, Barletta B, Palumbo S, Afferni C, Iacovacci P, Mari A, et al. Molecular characterization of a cross-reactive Juniperus oxycedrus pollen allergen, Jun o 2: a novel calcium-binding allergen. J Allergy Clin Immunol 1998;101:772-777

BACKGROUND: Species belonging to the Cupressaceae family are a relevant source of allergens that are present in a wide number of countries. OBJECTIVE: We sought to identify, purify, and characterize recombinant allergens from Juniperus oxycedrus, a species belonging to the Cupressaceae family. METHODS: Double-stranded cDNA was synthesized from mRNA and cloned into the lambda-ZAP expression vector. IgE screening of the library was performed with a pool of sera from subjects allergic to Cupressaceae. A recombinant 6xHis-tagged Juniperus oxycedrus allergen, Jun o 2, was expressed in Escherichia coli and purified by Ni2+ affinity chromatography. It was studied further by immunoblotting inhibition with pollen extracts from other Cupressaceae, Oleaceae, Urticaceae, and Graminaceae. The role of protein-bound calcium on the allergen's IgE-binding capacity was tested in a plaque assay in the presence or absence of EGTA. RESULTS: A cDNA coding for a newly identified Juniperus oxycedrus pollen allergen, rJun o 2, was isolated. The deduced amino acid sequence contained four typical Ca2+ binding sites and showed a significant sequence similarity to calmodulins. Depletion of Ca2+ in the plaque assay led to a loss of IgE-binding capacity of rJun o 2. Immunoblotting inhibition revealed that J. oxycedrus, J. ashei, Cupressus arizonica, C. sempervirens, Parietaria judaica, Olea europaea, and Lolium perenne pollen extracts were able to inhibit IgE binding to blotted rJun o 2 at different concentrations. CONCLUSION: rJun o 2 contains IgE-binding epitopes shared by taxonomically unrelated species, and therefore it can be regarded as a new panallergen. These findings could contribute to an explanation for the phenomenon of multiple positive test results in polysensitized patients and the potential symptom-eliciting role of allergenic sources previously not encountered.

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[73] - Pham NH, Baldo BA. Allergenic relationship between taxonomically diverse pollens. Clin Exp Allergy 1995;25:599-606

Skin tests and tests for IgE antibodies show that subjects are usually sensitive to a number of different pollens, frequently from taxonomically diverse species which are assumed to be allergenically non-crossreactive. This suggests that the presence of IgE antibody-reactivity to an individual pollen may not necessarily have resulted from contact with that pollen or even with a taxonomically closely related species. OBJECTIVE: Since this has important consequences for allergen avoidance and desensitization of patients, we attempted to define allergenic relationships between diverse pollen species. METHODS: Sera from subjects were examined in direct IgE antibody binding experiments and by quantitative inhibition, protein blotting and adsorption and elution studies. RESULTS: Sera from subjects diagnosed as allergic to white cypress pine, Italian cypress, ryegrass or birch pollen were shown to have IgE antibodies that reacted with pollens from these four species and from cocksfoot, couch grass, lamb's quarter, wall pellitory, olive, plantain and ragweed. These reactions were confirmed in protein blotting and adsorption and elution studies where numerous IgE-binding bands were detected in all 11 different pollen extracts with sera from each of the different allergic categories. Further evidence of allergenic (i.e. IgE-binding crossreactivity between the different pollens was provided by inhibition studies in which clear-cut inhibitions of IgE binding to the different pollen allergen discs were obtained with comparable amounts of the different pollen extracts. CONCLUSION: We conclude that the presence of pollen reactive IgE antibodies may not necessarily be a true reflection of sensitizing pollen species.

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[74] - 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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[75] - Sharma S, Panzani RC, Gaur SN, Ariano R, Singh AB. Evaluation of Cross-Reactivity between Holoptelea integrifolia and Parietaria judaica. Int Arch Allergy Immunol 2005;136:103-112

BACKGROUND: Holoptelea integrifolia and Parietaria judaica belong to the family Urticaceae, but are geographically distantly located. H. integrifolia is an important pollen allergen of India and sensitizes almost 10% of the atopic population in Delhi. P. judaica, on the other hand, is a very dominant pollen allergen of the Mediterranean region, sensitizing almost 80% of the allergic population. Since both these important pollen allergens belong to the family Urticaceae, the objective of the present study was to assess cross-reactivity between these two pollen allergens from different geographical regions . METHODS: Cross-reactivity between these two pollen allergens was assessed on the basis of skin prick tests and ELISA, ELISA inhibition and immunoblot inhibition studies . RESULTS: Out of 44 atopic Indian patients skin prick tested with H. integrifolia extract, 34% were found to be sensitized. All the patients sensitized to H. integrifolia also showed varying degrees of skin positivity to P. judaica pollen extract. ELISA and ELISA inhibition studies suggested strong cross-reactivity between H. integrifolia and P. judaica pollen. Immunoblot inhibition studies revealed that 14-, 16-, 28-, 38-, 42- and 46-kDa proteins are the cross-reactive proteins in H. integrifolia and P. judaica. However, Par j 1, the major allergen of P. judaica, is absent in H. integrifolia pollen . CONCLUSION: H. integrifolia and P. judaica pollens share cross-reactive as well as unique epitopes. The major allergen of P. judaica, Par j 1, seems to be absent in H. integrifolia pollen allergen.

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

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[77] - Asero R, Mistrello G, Roncarolo D, Amato S. Parietaria Profilin Shows Only Limited Cross-Reactivity with Birch and Grass Profilins. Int Arch Allergy Immunol 2004;133:121-124

BACKGROUND: In patients showing multiple reactivity to seasonal airborne allergens, a skin prick test (SPT) with Parietaria pollen scores frequently negative, suggesting that profilin from Parietaria might not share IgE-binding epitopes with profilin from botanically unrelated airborne allergens. This study investigated the immunologic cross-reactivity between profilins from Parietaria, grass and birch pollen . METHODS: 36 patients hypersensitive to birch profilin, Bet v 2, underwent SPT with Parietaria, and IgE to both whole Parietaria pollen and Phleum profilin were sought in their sera by ELISA. In ELISA inhibition studies, IgE reactivity to Phleum profilin was measured before and after absorption of sera from Parietaria reactors with both whole Parietaria pollen extract, and Par j 1/Par j 2 fraction. Further, their IgE reactivity to whole Parietaria pollen was measured before and after absorption with the Par j 1/Par j 2 fraction . RESULTS: All sera showed IgE reactivity to Phleum profilin. Only 14/36 (39%) patients had a positive SPT to Parietaria and 17/36 (47%) showed IgE to Parietaria. Absorption of sera from Parietaria reactors with whole Parietaria extract caused a marked reduction in IgE reactivity to grass profilin; in contrast, absorption of sera with the Par j 1/Par j 2 fraction did not exert any inhibitory effect on IgE reactivity to grass profilin. Absorption of sera with the Par j 1/Par j 2 fraction markedly reduced IgE reactivity to whole Parietaria extract in 8/9 cases . CONCLUSION: Less than 50% of patients sensitized to birch and grass profilin cross-react to Parietaria profilin. In most cases, cross-reactors are hypersensitive to major, specific Parietaria allergens as well. This findings may be of practical relevance when the prescription of specific immunotherapy is considered.

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[78] - Liccardi G, Mistrello P, Noschese P, Falagiani P, d'Amato M, d'Amato G. Oral allergy syndrome in pollinosis patients after eating pistachio nuts: two cases with different patterns of onset. Allergy 1996;51:919-922

We describe two uncommon cases of oral allergy syndrome (OAS) after eating pistachio nuts (p.n.) in subjects (a 54-year-old man and a 3-year-old girl) with exclusive skin prick test (SPT) positivity to Parietaria (P.) and pistachio nut (p.n.) allergens. Serologic P.- and p.n.-specific IgE determinations were also carried out. A double-blind, placebo-controlled food challenge (DBPCFC) was performed, for ethical reasons, only in the adult patient, but we observed a positive intraoral reaction only after slight scratching of the oral mucosa. Since this patient had put three whole p.n. with their shells into his mouth, breaking them with his teeth, before the onset of symptoms, we suggest that slight injury of the oral mucosa may enhance the local response. Preliminary results with SDS-PAGE and immunoblotting demonstrate the occurrence of a slight degree of cross-reactivity between these allergens, but further studies are necessary to obtain a definite conclusion.

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[79] - Liccardi G, Russo M, Mistrello P, Falagiani M, d'Amato M, d'Amato G. Sensitization to pistachio is common in Paretaria allergy. Allergy 1999;54:643-645

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[80] - Kanny G, Frémont S, Frentz P, Moneret-Vautrin DA. Allergie à la pistache: réaction croisée avec le pollen de pariétaire. Communication: 8èmes Journées Parisiennes d'Allergie - Paris 12-14 janvier 2000

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[81] - Minciullo PL, Mistrello G, Patafi M, Zanoni D, Gangemi S. Cross-reactivity between Parietaria pollen and beet. Allergol Immunopathol (Madr) 2007;35:74-75

Allergy to beet is very rare. Until now, only a few reports about asthma induced by inhaling the vapor of cooked beet have been published. We describe two patients with allergic rhinitis and positive skin prick tests to Parietaria and beet only. To investigate possible cross-reactivity between Parietaria pollen and beet, we performed laboratory assays that showed beet-specific IgE in the sera of both patients and possible cross-reactivity between Parietaria and beet in one patient.

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[82] - 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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[83] - Cirillo A, Patella V, Ciccarelli A, Leone G, Gallo L. Food allergy to rocket (Eruca sativa) in patients sensitizited to parietaria pollen. 8th International Symposium on Problems of Food Allergy, Venice 2001, March 11-13

Introduction. In southern Italy respiratory allergy to Parietaria pollen is very common. In our Allergy Unit in the current year 240 patients (178 males, 62 females, with a mean age of 24.3 ± 8.4 years) were diagnosed as monosensitised to Parietaria pollen. Some of these patients reported also allergic reactions to foods, and four of them identified the culprit food as rocket (Eruca sativa), a vegetable belonging to the Cruciferae family increasingly used in a number of food preparations Methods. Skin tests were done by the prick + prick technique with fresh rocket, since commercial extracts are not available. Clinical cases. The four patients were three females respectively 23, 24 and 27 years old, and one male 31 year old. All had rhinoconjunctivitis and/or asthma in the pollen season of Parietaria, while the allergic reactions to the ingestion of rocket were urticaria in two patients, urticaria and asthma in one patient, and asthma alone in one patients. Results. The skin responses to the prick + prick with fresh rocket were intensely positive in all patients, being graded in 3+/4+ reactions in respect to the reference positive wheal elicited by the histamine solution. Conclusions. The vegetable food Eruca sativa was so far very rarely reported as a cause of food allergy, and most case reports regard contact urticaria. The cases we describe demonstrate that this food can elicit different patterns of systemic reactions, involving the skin or the airways, and that such reactions are not so rare in subjects suffering from Parietaria pollinosis. This suggests the presence of cross-reacting allergens in the two sources.

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[84] - Pucci S, Incorvaia C. Allergic reactions to eggplant in subjects with parietaria pollinosis. 8th International Symposium on Problems of Food Allergy, Venice 2001, March 11-13

Introduction. Cross-reactions between pollens and foods caused by the presence of highly homologous allergens in the different sources are well known, and define the birch-apple syndrome, the mugwort-celery syndrome, the grass-tomato-peanut syndrome, and others (1,2). We report a new clinical association between Parietaria pollinosis and allergic reactions to eggplant. Material and methods. Five patients, three women and two men, of age between 24 and 50 years, with allergy to ingestion of eggplants are reported. All suffered from Parietaria pollinosis from a time length ranging from 5 to 20 years and had recently had allergic reactions to ingestion of eggplant, characterised in three cases by symptoms of OAS (one only oropharyngeal symptoms, one also cough and dysphonia, one vomiting), but in two cases by systemic symptoms too (one urticaria, one anaphylactic shock). These patients were tested by skin prick tests with commercial pollen extracts and by the prick + prick technique with a standard panel of fresh fruits and vegetables. Results. Skin prick tests with commercial pollen extracts were positive (with a 4+ grading) to Parietaria in all patients and also to grass pollen and Oleaceae (1+) in one patient and to grass pollen and Cupressaceae (1+) in another. No symptoms in the pollination season of these plants were reported. Prick + prick were positive to eggplant (4+) in all patients, to almond (3+) in one patient, and to fennel (2+) and basil (2+) in another. Only the sensitisation to fennel was clinically expressed with OAS symptoms. Conclusions. The clear relationship between Parietaria pollinosis and allergy to eggplant allows to define a new association, the Parietaria- eggplant syndrome. The fact that the studied patients had respiratory symptoms only to Parietaria and also a limited number of sensitisations to fruits and vegetables (of which only that to fennel was clinically expressed) suggests that a specific allergen, and not one of the already} mown panallergens (2), often involved in the pollen-vegetable syndromes, is responsible. The nature of such allergen has to be investigated

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[85] - Miralles JC, Garcia-Sellés J, Bartolomé B, Negro JM. Occupational rhinitis and bronchial asthma due to artichoke (Cynara scolymus). Ann Allergy Asthma Immunol 2003;91:92-95

BACKGROUND: The artichoke is a perennial horticultural plant that belongs to the Compositae family. OBJECTIVE: To present case studies of 2 vegetable warehouse workers who developed occupational rhinitis and bronchial asthma by sensitization to artichoke. METHODS: Skin prick tests with common inhalants and foods were performed. Specific IgE to artichoke, Parietaria judaica pollen, and Olea europaea pollen extracts was measured by a specific IgE enzyme immunosorbent assay kit. Molecular mass of the allergens was studied by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) immunoblotting technique. Patients underwent a nasal challenge test, and one patient provided peak expiratory flow rate (PEFR) measurements in her workplace. RESULTS: In both patients, results of skin prick tests to artichoke were positive. Levels of specific IgE for artichoke were 0.68 kU/L in patient 1 and 2.14 kU/L in patient 2. The protein composition of the artichoke extract, studied by SDS-PAGE, showed that most bands ranged from 30 to 14 kDa. The IgE-binding bands with the serum samples of patient 1 showed apparent molecular masses of 56, 48, 38, 31, 27, 25, 16, and 15 kDa; however, the serum samples of patient 2 showed IgE bands of 21 and 19 kDa. Western blotting of artichoke extract showed a complete inhibition of IgE-binding bands when serum samples were preincubated with P. judaica pollen extract. Nasal challenge with artichoke extract triggered a peak nasal inspiratory flow decrease of 81% and 85% in patient 1 and patient 2, respectively. Finally, patient 1 recorded a PEFR decrease of up to 36% after exposure to artichoke in her workplace. CONCLUSIONS: SDS-PAGE immunoblotting inhibition performed for the artichoke extract showed a total disappearance of the specific IgE binding bands when serum samples were previously incubated with P. judaica pollen extract, thus establishing the existence of a serologic cross-reactivity between artichoke and P. judaica pollen.

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[86] - Ibero M, Castillo M, Viñas M, Bartolomé B. Sensitization to Non-Specific Lipid Transfer Protein from Parietaria Judaica Pollen and Cross Sensitization to Foods. AAAAI 62nd Annual Meeting, Miami, 3-7 March 2006, Poster n°187

CASE REPORT: A 16-year-old boy, diagnosed of rhinoconjunctivitis and sensitizated to: dust, molds, cat epithelium, dog epithelium and pollens (Olea europea, Cupressus arizonica, Lolium perenne, Cynodon dactylon, Parietaria judaica and Plantago lanceolata), that suffered from anaphylaxis after eating chicken with potatoes and artichoke. Several months later the patient suffered from swelling of the lips with almonds Later he has tolerated chicken and potatoes METHODS AND RESULTS: Skin prick tests were positive to extracts from: artichoke (crude and cooked) and nuts. Specific IgE (EAST) were: Flower of Artichoke (flower+head-food): 8.6 kU/l; Stem of Artichoke: 3.6 kU/l; Artichoke (head-food): 2.9 kU/l; peanut: 26.6 kU/l and almonds: 2.23 kU/l.. SDS-PAGE Immunoblotting with artichoke extract showed IgE-binding bands at 14 kDa, 15,5 kDa and 17 kDa. Cross reactivity was studied between the artichoke extract and the extract from these pollens to which the patient was sensitizated. EAST inhibition using artichoke extract as solid phase showed an inhibition of 60% with P. judaica pollen extract. Immunoblotting with artichoke extract showed a complete inhibition of the IgE-binding when patient serum was preincubated with P. judaica pollen extract. Inhibition studies showed the artichoke IgEbinding proteins cross-react with the lipid transfer protein (LTP) from peach CONCLUSIONS: In our knowledge it is the first case of oral allergy to artichoke. The clinical evolution of the patient and the in vitro test suggest an initial sensitization by respiratory tract to pollen from P. judaica, and a later cross sensitization by digestive tract to artichoke.

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[87] - Castillo M, Ibero M, Vinas M, Bartolome B. Cross Sensitization between Parietaria judaica Pollen and Vegetal Foods. J Allergy Clin Immunol 2008;121:S184

RATIONALE: Some researchers demonstrated that sensitization to various pollens predispose to suffer from food allergy. We decided to investigate the relation between the sensitization to Parietaria judaica pollen and the subsequent develop of food allergy. METHODS: We recruited 27 patients allergic only or in major degree to Parietaria judaica pollen who underwent a protocol consisting on: cutaneous tests to extracts from Parietaria judaica pollen, artichoke, hazelnut, peanut, pistachio, peach peel and peach pulp; clinical history on adverse reactions with peach, nuts or artichoke and specific IgE to Parietaria judaica pollen, artichoke (flower, stem and leaves), hazelnut, peanut, peach (peel and lipid transfer protein (LTP)) and almond (roasted and raw). Finally we performed SDS-PAGE immunoblotting with Parietaria judaica pollen extract. RESULTS: All patients had serum specific IgE to Parietaria judaica pollen (class 2 to 4) without significant differences in the IgE levels to Par j 1 and Par j 2. Nine from 27 (33.3%) of them had clinical and/or positive tests and/or clinical allergic history to peach, nuts or artichoke. All patient sera revealed the same IgE-binding bands at 16, 29, 40 and 55 kDa; all of these bands turned to be different molecular forms of the major allergens (Par j 1 and Par j 2). CONCLUSIONS: As the main allergens from Parietaria judaica pollen are non-specific LTP, the fact that only some of the P. judaica pollen allergic patients develop food allergy, could be explained in terms of Par j 1, Par j 2 specific IgE concentration, differences on epitope recognition or both mechanisms.

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[88] - Fedele R, Ricciardi L, Tigano V, Amabile A, Isola S, Patafi M, et al. Cross-reactivity between parietaria pollen and mulberry fruit. EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°653

Background: The presence of cross-reactivity between pollens and vegetal foods is well known. As a patient affected by rhinitis and asthma from parietaria had referred to us because of an anaphylactic shock after the ingestion of mulberries, we carried out a study in order to evaluate the cross-reactivity between parietaria pollen and mulberry fruit. Method: 44 patients allergic to parietaria pollen (28 f and 16 m), aged between 11 and 65 years, and 35 atopic patients but not allergic to parietaria (18 f and 17 m), aged between 16 and 58 years, underwent a prick by prick with mulberries. Results: 31 patients (19 f and 12 m) aged between 11 and 65 years were positive for mulberry while the other 19 patients (9 f and 4 m), aged between 12 and 47 years, were negative. Four of the 31 patients positive to mulberry fruit, apart from the patient who had reported the anaphylactic shock, had presented an allergic reaction after eating this fruit: oral allergic syndrome in 2 cases, associated to rhinoconjunctivitis in another case, oedema of the tongue in one case. None of the patients negative to the prick by prick test with mulberries had ever experienced an allergic reaction after eating this fruit. It is important to notice that 20 of the 31 patients positive to mulberry fruit denied to have ever eaten it. Of the 35 control atopic patients only 3 had a mild positivity to the prick by prick test with mulberry; these patients reported to have eaten this fruit without developing any reaction. Conclusion: The presence of common antigenic determinants between parietaria pollen and mulberries may explain the frequent association found between allergy to parietaria and skin positivity to mulberry fruit, even in patients who reported not to have ever eaten these fruits. As some of the patients who resulted positive to mulberries had experienced an allergic reaction after eating these fruits, further studies are still needed in order to find these common antigenic determinants and to evaluate the real incidence of allergy to mulberries in patients allergic to parietaria pollen.

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[89] - Asero R, Mistrello G, Roncarolo D, Amato S, Caldironi G, Barocci F, et al. Immunological cross-reactivity between lipid transfer proteins from botanically unrelated plant-derived foods: a clinical study. Allergy 2002;57:900-906

BACKGROUND: Lipid transfer proteins (LTP) are highly conserved and widely distributed throughout the plant kingdom. Recent studies demonstrated immunological cross-reactivity between LTP from many botanically unrelated fruits and vegetables and concluded that LTP are pan-allergens. This study aimed to evaluate the clinical relevance of such cross-reactivity in a group of subjects monosensitized to LTP . METHODS: Twenty LTP-hypersensitive patients were selected from a population of about 600 subjects with history of Rosaceae allergy by means of: 1) negative skin prick test (SPT) with a commercial birch pollen extract; 2) positive SPT with a commercial plum extract, rich in LTP but virtually lacking both Bet v 1-like proteins and profilin; 3) in-vitro IgE reactivity to the 9-10 kDa fraction of peach peel or immunoblot with peach peel showing a single band at 10 kDa; and 4) total inhibition of reactivity to whole peach extract (containing Bet v 1-related allergen, profilin, and LTP) by purified peach LTP on enzyme-linked immunoassay (ELISA). Allergy to foods other than Rosaceae was ascertained by careful interview and analysis of medical recordings. SPT with a large series of plant-derived foods were carried out as well. The cross reactivity between LTPs from botanically unrelated plant-derived foods was assessed by ELISA inhibition tests using walnut and peanut extracts as substrate, and peach LTP as inhibitor . RESULTS: All patients reported allergic reactions after the ingestion of at least one from a large number of vegetable foods other than Rosaceae, and in several cases clinical reactions were very severe (anaphylaxis, asthma, urticaria/angioedema). Nuts and peanuts were the most frequently reported causes of allergic reactions (80% and 40% of patients, respectively). All patients showed positive SPT to several non-Rosaceae food extracts. SPT with nuts, peanut, legumes, celery, rice, and corn were positive in the majority of patients. In ELISA inhibition studies, absorption of sera with peach LTP caused complete inhibition of IgE reactivity to walnut and peanut in all cases . CONCLUSION: LTP is a clinically relevant pan-allergen. Most Rosaceae-allergic, LTP-hypersensitive patients experience adverse reactions after ingestion of botanically unrelated plant-derived foods as well. In view of the high prevalence and severity of the allergic reactions induced, hazelnut, walnut, and peanut should be regarded as potentially hazardous for these patients.

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Les allergènes des pariétaires

Réactions croisées des pariétaires avec d’autres pollens

Réactions croisées des pariétaires avec des aliments végétaux

Rubriques

Dans la même rubrique

Les Amaranthacées

Les ambroisies

L’armoise

Les Astéracées (Composées)

Les plantains

Les Euphorbiacées