Les plantains

[1] - 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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[2] - Laaidi K, Besancenot JP, Carli PM. Evolution de l'éosinophilie au cours de la saison pollinique dans une population générale: un outil pour déterminer de nouvelles sensibilisations. Allerg Immunol (Paris) 2002;34:13-18

The purpose of this paper was to verify the effect of pollen peaks on blood eosinophilia in an all and sundry population, including allergic as well as non-allergic subjects, so that we can open up new horizons in the understanding and prevention of pollinosis. Daily eosinophilia counts of hospital patients were measured at the time of a blood checkup (1996-1998), and divided into six classes. Those data were compared to daily pollen counts of twelve taxa, coming from the Hirst trap of Dijon (France). An eosinophilia increase occurred when hazel, hornebeam, birch, oak, grasses, ragweed and plantain were present in high concentration. In other cases, only simultaneous presence of several taxa seemed to play a part, because of cross-reactivity or polysensitization. Lastly, Cupressaceae-Taxaceae and ragweed were seen as increasing eosinophilia in seemingly non allergic people. The analysis of eosinophilia in the general population was able to reveal potential allergic patients and potential allergic diseases.

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[3] - Mehta V, Wheeler AW. IgE-mediated sensitization to English plantain pollen in seasonal respiratory allergy: identification and partial characterisation of its allergenic components . Int Arch Allergy Appl Immunol 1991;96:211-217

Characterisation by SDS-PAGE immunoblotting of plantain pollen extract showed that components of 16,000-20,000 M(r) were frequently reactive with IgE antibody in the sera of subjects with seasonal respiratory allergy. Other, more weakly IgE-binding allergens were seen in the range of 40,000-60,000 M(r). HPLC followed by RAST inhibition demonstrated that components of approximately 17,000 M(r) were also responsible for much of the IgE-binding activity of the extract. These components appeared to have pI values between 4.5 and 5.2. RAST inhibition showed that there were no common IgE-binding epitopes in grass pollen and plantain pollen extracts, indicating that skin test responses should not necessarily be interpreted in terms of cross-reaction. 82 subjects with a clinical history of seasonal, respiratory allergy were screened in a skin prick test survey. 28% were skin test positive to plantain pollen extract. The frequency of positive skin test reactions to plantain pollen extract was greater than that to Betula (23%) and Artemisa (16%), both which are considered to be important allergens. In a larger survey positive RAST scores to plantain pollen were given by 34% of sera from subjects with respiratory allergy. Plantain pollen sensitivity should therefore be considered during diagnosis of seasonal allergy

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

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

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[5] - Cuesta-Herranz J, Lazaro M, Figueredo E, Igea JM, Umpierrez A, De Las Heras M. Allergy to plant-derived fresh foods in a birch- and ragweed-free area. Clin Exp Allergy 2000;30:1411-1416

BACKGROUND: Allergy to plant-derived fresh foods has often been reported in geographical areas where birch or ragweed pollens are frequent and has been attributed to cross-reactivity to pollens. OBJECTIVE: The aim of this study has been to evaluate allergy to plant-derived fresh foods among pollen-allergic patients from a birch and ragweed-free area. METHODS: Ninety-five pollen-allergic patients took part in the study. The study consisted of a questionnaire, skin prick tests and challenge tests. Pollen skin tests to five grasses, eight trees and seven weeds were performed in duplicate. Prick tests (prick by prick) and challenge tests were carried out with the fresh foods. RESULTS: Most patients allergic to pollens were sensitized to grass (Lolium and Phleum; 97.9%), followed by tree (Olea; 82.1%) and weed pollens (Plantago; 64.2%). 35 of the 95 pollen-allergic patients had positive skin test responses to some plant-derived fresh foods, the highest percentage corresponding to several fruits in the Rosaceae family (peach and pear, 26.3%), followed by Cucurbitacea fruits (melon, 13.7%). The 21. 05% of the pollen-allergic patients were allergic to some type of plant-derived fresh food. Peach was the plant-derived fresh food which most frequently elicited allergy symptoms (12.6%), followed by melon (7.36%). The cluster of positive responses to Rosaceae fruits was higher for skin testing than for challenge testing. CONCLUSION: Peach was the most important allergy provoking fruit in a birch and ragweed free-area where apples were consumed at a rate of two times more than peaches and the patients allergic to pollen were principally sensitized to grass pollens.

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[6] - 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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[7] - Alemán A, Sastre J, Quirce S, de las Heras M, Carnés J, Fernández-Caldas E, et al. Allergy to kiwi: A double-blind, placebo-controlled food challenge study in patients from a birch-free area. J Allergy Clin Immunol 2004;113:543-550

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

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

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

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[9] - Calabozo B, Barber D, Polo F. Purification and characterization of the main allergen of Plantago lanceolata pollen, Pla l 1. Clin Exp Allergy 2001;31:322-330

English plantain (Plantago lanceolata) pollen is an important cause of pollinosis in the temperate regions of North America, Australia and Europe. However, very little is known about its allergen composition. The aim of this study was to identify plantain allergens, and to isolate and characterize a major allergen. Allergens were identified by immunoblotting with individual allergic patients' sera. Isolation of the major allergen was achieved by sequential reverse-phase and size-exclusion HPLC. Allergenic characterization was performed by ELISA and immunoblotting after SDS-PAGE with sera from plantain-allergic patients. N-terminal amino acid sequence was established by Edman degradation. Allergograms showed that 13 out of the 14 sera assayed had IgE to a group of proteins with a molecular weight in the range of 16-20 kd, that turned out to be different isoforms or variants of the major allergen Pla l l. Eighteen amino acid residues from the N-terminal end of one of the isoforms, and 10 of three others, were sequenced, and a partial sequence identity with Ole e 1 was found. Prevalence of specific IgE to purified Pla l 1 in plantain allergic patients was 86%, and represents about 80% of the total IgE-binding capacity of the plantain extract. The most relevant allergen from P.lanceolata pollen, Pla l 1, has been purified and characterized. This contributes to a greater knowledge of the allergen composition of this important weed, and clears the way for the standardization of plantain allergen products in terms of major allergen content.

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[10] - 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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[11] - Nakamaru Y, Maguchi S, Oridate N, Takagi D, Furuta Y, Fukuda S. Plantago lanceolata (English plantain) pollinosis in Japan. Auris Nasus Larynx 2005;32:251-256

OBJECTIVE: The objective was to ascertain the prevalence of Plantago lanceolata (English plantain) pollinosis in Japan. METHODS: A total of 160 patients with allergic rhinitis were examined at the Hokkaido University Hospital Otolaryngology Clinic between January 2002 and December 2003. We investigated the frequency of P. lanceolata antigen-specific immunoglobulin E (IgE) antibody-positive serum using the radioallergosorbent test (RAST), the P. lanceolata pollen counts in the Sapporo area and the clinical symptoms of patients with allergic rhinitis caused by this pollen. RESULTS: P. lanceolata pollen was dispersed throughout the Sapporo area from mid-May to early September, peaking in the latter half of May. The airborne pollen count showed little variation between years. The P. lanceolata RAST-positive rate was 12.8%, which was close to those for ragweed and sagebrush. All subjects showed overlapping antigen sensitization, that is, none displayed sensitization (by RAST) exclusively to P. lanceolata. The onset of subjective symptoms peaked in June and the symptoms worsened from April to October. CONCLUSION: P. lanceolata was dispersed over a long period, from mid-May to early September. The P. lanceolata RAST-positive rate (12.8%) was similar to those for sagebrush and ragweed, which are dispersed during the fall. These results suggest that P. lanceolata is an important causative agent of pollinosis in Japan.

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

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

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

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[14] - Mohapatra SS, Lockey RF, Polo F. Weed pollen allergens. Clin Allergy Immunol 2004;18:207-222

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[15] - Antoine LC, Dubreil Y, Galaup B, Leduc V. Mono-sensibilisation au pollen de plantain (Plantago lanceolata). Rev Fr Allergol Immunol Clin 2007;47:272

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[16] - Wojdas A, Rapiejko P, Zielnik-Jurkiewicz B, Kantor I. Nasal provocation test in patients allergic to pollen. Ann Agric Environ Med 2005;12:173-176

Nasal provocative test (NPT) can be defined as a method for recreating upper respiratory tract response to natural allergens or irritants. It can be used in solving nasal patophisiology problems: establishing whether and how the patient's nose is sensitive to antigens or irritants; quantitative evaluation of sensitivity; establishing factors influencing sensitivity. This method is employed to confirm clinical diagnosis in cases where difficulties arise in interpretation of diagnostic tests. The study based on nasal provocative tests establish an allergy to pollen in cases of pollinosis, and select appropriate components for the desensitising vaccine. Sample group included 53 patients, 29 were females and 24 were males, aged 15-42 years, selected from 1,021 patients diagnosed between 1999-2002 in the Allergology Department of the ENT Department of the MMI. The sample patients were diagnosed based on additional tests with allergic inflammation of the nasal mucosa caused by allergy to pollen of such plants as birch, grass, rye, mugwort and plantain. Research methods included: subjective physical examination, prick tests, total and specific IgE levels in serum, nasal provocative tests and rhinomanometric examination. Allergen solution was administered onto the mucosa with a calibrated atomiser. NPT solutions containing pollen of birch, grass, rye, mugwort and plantain were used. Provocative test was considered positive if, following allergen provocation, rhinomanometric examination revealed an increase in respiratory resistance by at least 40 % in comparison with the control test. On the basis of the study, 2 conclusions were drawn: 1) Nasal provocative test is an essential element in diagnostics of allergic nasal obstruction. 2) Rhinomanometry, as an objective method of examining nasal patency, is crucial for evaluating the nasal provocative test.

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[17] - Antoine LC, Dubreil Y, Galaup B, Leduc V. Mono-sensibilisation au pollen de plantain (Plantago lanceolata). Rev Fr Allergol Immunol Clin 2007;47:272

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[18] - Mohapatra SS, Lockey RF, Polo F. Weed pollen allergens. In Lockey RF. and Ledford DK, (eds). Allergens and Allergen Immunotherapy for Allergic Diseases. 4th ed. 2008, New York: Informa Healthcare USA, Inc., p. 127-39

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[19] - Calabozo B, Barber D, Polo F. Purification and characterization of the main allergen of Plantago lanceolata pollen, Pla l 1. Clin Exp Allergy 2001;31:322-330

English plantain (Plantago lanceolata) pollen is an important cause of pollinosis in the temperate regions of North America, Australia and Europe. However, very little is known about its allergen composition. The aim of this study was to identify plantain allergens, and to isolate and characterize a major allergen. Allergens were identified by immunoblotting with individual allergic patients' sera. Isolation of the major allergen was achieved by sequential reverse-phase and size-exclusion HPLC. Allergenic characterization was performed by ELISA and immunoblotting after SDS-PAGE with sera from plantain-allergic patients. N-terminal amino acid sequence was established by Edman degradation. Allergograms showed that 13 out of the 14 sera assayed had IgE to a group of proteins with a molecular weight in the range of 16-20 kd, that turned out to be different isoforms or variants of the major allergen Pla l l. Eighteen amino acid residues from the N-terminal end of one of the isoforms, and 10 of three others, were sequenced, and a partial sequence identity with Ole e 1 was found. Prevalence of specific IgE to purified Pla l 1 in plantain allergic patients was 86%, and represents about 80% of the total IgE-binding capacity of the plantain extract. The most relevant allergen from P.lanceolata pollen, Pla l 1, has been purified and characterized. This contributes to a greater knowledge of the allergen composition of this important weed, and clears the way for the standardization of plantain allergen products in terms of major allergen content.

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[20] - Mohapatra SS, Lockey RF, Polo F. Weed pollen allergens. Clin Allergy Immunol 2004;18:207-222

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[21] - Calabozo B, Diaz-Perales A, Salcedo G, Barber D, Polo F. Cloning and expression of biologically active Plantago lanceolata pollen allergen Pla l 1 in the yeast Pichia pastoris. Biochem J 2003;372:889-896

The glycoprotein Pla l 1 is the major allergen from Plantago lanceolata (English plantain) pollen, which is a common cause of pollinosis in temperate areas. Three complete cDNAs for Pla l 1 isoforms were isolated by polymerase chain reactions using specific 3' and 5' primers. All three Pla l 1 cDNAs code for a 25-residue leader peptide and a 131-residue mature protein that contains two polymorphic positions, an N-glycosylation site at position 107, and six cysteine residues involved in three disulfide bridges. The allergen variant Pla l 1.0101 was produced in P.pastoris at a yield of 20 mg per litre of culture as a mixture of non-glycosylated (17 kDa), glycosylated (23 kDa) and dimeric forms (32-39 kDa). rPla l 1 was purified by affinity chromatography with an anti-nPla l 1 monoclonal antibody, and its molecular and immunological properties were compared with the natural allergen by circular dichroism (CD) analysis, enzymatic deglycosylation, lectin binding assay, immunodetection and ELISA-inhibition assays using sera from plantain-allergic patients. The recombinant allergen is properly folded as deduced from CD spectra, and the immunodominant allergenic epitopes of the natural allergen are preserved in rPla l 1. These results allow to conclude that P.pastoris is a convenient system for the efficient production of biologically active rPla l 1, which could have a potential use for clinical purposes. Furthermore, a sequence similarity of Pla l 1 with the major allergen from the olive tree pollen, Ole e 1, is disclosed in this work, and the allergenic cross-reactivity between both allergens has been studied.

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[22] - Calabozo B, Barber D, Polo F. Studies on the carbohydrate moiety of Pla l 1 allergen. Identification of a major N-glycan and significance for the immunoglobulin E-binding activity. Clin Exp Allergy 2002;32:1628-1634

Background Pla l 1, the major allergen of Plantago lanceolata pollen, is a glycoprotein that contains an N-glycosylation site. Carbohydrate moieties of many allergenic glycoproteins have been reported to be IgE-binding determinants responsible for cross-reactivity among different species. Objective : To identify the kind of linkages and the type of glycans present in Pla l 1 and to investigate their contribution to the allergic response to this allergen. Method s : Pla l 1 was deglycosylated by N-glycosidase A and the IgE-binding ability of the unglycosylated protein was evaluated by dot-blot. Identification of 1 2 xylose and/or 1 3 fucose residues in Pla l 1 N-glycan was carried out by incubation with specific antibodies from rabbit antiserum against HRP (anti-HRP). The contribution of this N-glycan to total IgE reactivity was analysed quantitatively by pre-incubation of Pla l 1 with anti-HRP prior to incubation with sera. The role of the carbohydrate moiety of Pla l 1 in cross-reactivity was studied by RAST using unrelated glycoproteins with known sugar composition and structure. Result s : The effectiveness of N-glycosidase A to deglycosylate Pla l 1 and the ineffectiveness of the treatment with PNGase F indicate that Pla l 1 carries a complex N-glycan with an 1 3 fucose residue in its structure. Furthermore, the presence of 1 2 xylose and/or 1 3 fucose residues was identified in this N-glycan by means of an ELISA. Pre-incubation of Pla l 1 with an anti-HRP antibody caused a weak but significant reduction in IgE reactivity. Some sera from P. lanceolata-allergic patients reacted positively with four glycoproteins that bear N-glycans of complex type but not with fetuine. Conclusions : Pla l 1 is a glycoprotein that carries at least a complex, major N-linked glycan, with a 1 3 fucose residue in its structure and probably also a 1 2 xylose. This glycan moiety does not seem to constitute a relevant allergenic epitope of Pla l 1.

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[23] - Asero R, Mistrello G, Roncarolo D, Casarini M. Detection of allergens in plantain (Plantago lanceolata) pollen. Allergy 2000;55:1059-1062

BACKGROUND: Allergens in Plantago lanceolata have not been characterized yet. The objective was to characterize some plantain-pollen allergens and to investigate the cross-reactivity between plantain and grass pollens. METHODS: Sera from four patients monosensitive to plantain pollen and from eight grass-pollen-allergic patients showing strong skin reactivity to plantain pollen in the skin prick test (SPT) underwent immunoblot analysis with both Plantago and grass mix extract. Moreover, immunoblot inhibition experiments were done with grass mix extract as inhibitor. RESULTS: All four sera from plantain-allergic patients reacted to two distinct bands at 17 and 19 kDa, and 2/4 sera showed further reactivity to a 40-kDa protein, which in one case represented the most prominent IgE-binding allergen. Plantain-monosensitive subjects did not show any reactivity to grass-pollen extract, and preabsorption of their sera with grass-pollen extract did not cause any loss of reactivity to plantain pollen. Sera from all eight grass-pollen-allergic controls reacted to a 30-kDa protein in plantain pollen, and some sera showed cross-reactivity to higher and lower molecular-weight structures as well. In all cases, plantain reactivity was totally abolished by preabsorption of sera with grass-pollen extract. A preliminary investigation by immunoblot showed that polyclonal IgG anti-Phl p 5 (but not polyclonal Phl p 1) from rabbit reacted to a 30-kDa protein in plantain pollen. CONCLUSIONS: Three specific allergens (of 17, 19, and 40 kDa, respectively) have been detected in plantain pollen. Further studies on a larger number of patients will determine whether these proteins may be considered major allergens. Cross-reactivity between grass and plantain pollen is mainly caused by a 30-kDa protein in plantain pollen. Group 5 grass-pollen allergen is probably responsible for most grass/plantain cross-reactivity.

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[24] - Calabozo B, Diaz-Perales A, Salcedo G, Barber D, Polo F. Cloning and expression of biologically active Plantago lanceolata pollen allergen Pla l 1 in the yeast Pichia pastoris. Biochem J 2003;372:889-896

The glycoprotein Pla l 1 is the major allergen from Plantago lanceolata (English plantain) pollen, which is a common cause of pollinosis in temperate areas. Three complete cDNAs for Pla l 1 isoforms were isolated by polymerase chain reactions using specific 3' and 5' primers. All three Pla l 1 cDNAs code for a 25-residue leader peptide and a 131-residue mature protein that contains two polymorphic positions, an N-glycosylation site at position 107, and six cysteine residues involved in three disulfide bridges. The allergen variant Pla l 1.0101 was produced in P.pastoris at a yield of 20 mg per litre of culture as a mixture of non-glycosylated (17 kDa), glycosylated (23 kDa) and dimeric forms (32-39 kDa). rPla l 1 was purified by affinity chromatography with an anti-nPla l 1 monoclonal antibody, and its molecular and immunological properties were compared with the natural allergen by circular dichroism (CD) analysis, enzymatic deglycosylation, lectin binding assay, immunodetection and ELISA-inhibition assays using sera from plantain-allergic patients. The recombinant allergen is properly folded as deduced from CD spectra, and the immunodominant allergenic epitopes of the natural allergen are preserved in rPla l 1. These results allow to conclude that P.pastoris is a convenient system for the efficient production of biologically active rPla l 1, which could have a potential use for clinical purposes. Furthermore, a sequence similarity of Pla l 1 with the major allergen from the olive tree pollen, Ole e 1, is disclosed in this work, and the allergenic cross-reactivity between both allergens has been studied.

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[25] - Calabozo B, Barber D, Polo F. Purification and characterization of the main allergen of Plantago lanceolata pollen, Pla l 1. Clin Exp Allergy 2001;31:322-330

English plantain (Plantago lanceolata) pollen is an important cause of pollinosis in the temperate regions of North America, Australia and Europe. However, very little is known about its allergen composition. The aim of this study was to identify plantain allergens, and to isolate and characterize a major allergen. Allergens were identified by immunoblotting with individual allergic patients' sera. Isolation of the major allergen was achieved by sequential reverse-phase and size-exclusion HPLC. Allergenic characterization was performed by ELISA and immunoblotting after SDS-PAGE with sera from plantain-allergic patients. N-terminal amino acid sequence was established by Edman degradation. Allergograms showed that 13 out of the 14 sera assayed had IgE to a group of proteins with a molecular weight in the range of 16-20 kd, that turned out to be different isoforms or variants of the major allergen Pla l l. Eighteen amino acid residues from the N-terminal end of one of the isoforms, and 10 of three others, were sequenced, and a partial sequence identity with Ole e 1 was found. Prevalence of specific IgE to purified Pla l 1 in plantain allergic patients was 86%, and represents about 80% of the total IgE-binding capacity of the plantain extract. The most relevant allergen from P.lanceolata pollen, Pla l 1, has been purified and characterized. This contributes to a greater knowledge of the allergen composition of this important weed, and clears the way for the standardization of plantain allergen products in terms of major allergen content.

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[26] - Calabozo B, Barber D, Polo F. Studies on the carbohydrate moiety of Pla l 1 allergen. Identification of a major N-glycan and significance for the immunoglobulin E-binding activity. Clin Exp Allergy 2002;32:1628-1634

Background Pla l 1, the major allergen of Plantago lanceolata pollen, is a glycoprotein that contains an N-glycosylation site. Carbohydrate moieties of many allergenic glycoproteins have been reported to be IgE-binding determinants responsible for cross-reactivity among different species. Objective : To identify the kind of linkages and the type of glycans present in Pla l 1 and to investigate their contribution to the allergic response to this allergen. Method s : Pla l 1 was deglycosylated by N-glycosidase A and the IgE-binding ability of the unglycosylated protein was evaluated by dot-blot. Identification of 1 2 xylose and/or 1 3 fucose residues in Pla l 1 N-glycan was carried out by incubation with specific antibodies from rabbit antiserum against HRP (anti-HRP). The contribution of this N-glycan to total IgE reactivity was analysed quantitatively by pre-incubation of Pla l 1 with anti-HRP prior to incubation with sera. The role of the carbohydrate moiety of Pla l 1 in cross-reactivity was studied by RAST using unrelated glycoproteins with known sugar composition and structure. Result s : The effectiveness of N-glycosidase A to deglycosylate Pla l 1 and the ineffectiveness of the treatment with PNGase F indicate that Pla l 1 carries a complex N-glycan with an 1 3 fucose residue in its structure. Furthermore, the presence of 1 2 xylose and/or 1 3 fucose residues was identified in this N-glycan by means of an ELISA. Pre-incubation of Pla l 1 with an anti-HRP antibody caused a weak but significant reduction in IgE reactivity. Some sera from P. lanceolata-allergic patients reacted positively with four glycoproteins that bear N-glycans of complex type but not with fetuine. Conclusions : Pla l 1 is a glycoprotein that carries at least a complex, major N-linked glycan, with a 1 3 fucose residue in its structure and probably also a 1 2 xylose. This glycan moiety does not seem to constitute a relevant allergenic epitope of Pla l 1.

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[27] - Calabozo B, Diaz-Perales A, Salcedo G, Barber D, Polo F. Cloning and expression of biologically active Plantago lanceolata pollen allergen Pla l 1 in the yeast Pichia pastoris. Biochem J 2003;372:889-896

The glycoprotein Pla l 1 is the major allergen from Plantago lanceolata (English plantain) pollen, which is a common cause of pollinosis in temperate areas. Three complete cDNAs for Pla l 1 isoforms were isolated by polymerase chain reactions using specific 3' and 5' primers. All three Pla l 1 cDNAs code for a 25-residue leader peptide and a 131-residue mature protein that contains two polymorphic positions, an N-glycosylation site at position 107, and six cysteine residues involved in three disulfide bridges. The allergen variant Pla l 1.0101 was produced in P.pastoris at a yield of 20 mg per litre of culture as a mixture of non-glycosylated (17 kDa), glycosylated (23 kDa) and dimeric forms (32-39 kDa). rPla l 1 was purified by affinity chromatography with an anti-nPla l 1 monoclonal antibody, and its molecular and immunological properties were compared with the natural allergen by circular dichroism (CD) analysis, enzymatic deglycosylation, lectin binding assay, immunodetection and ELISA-inhibition assays using sera from plantain-allergic patients. The recombinant allergen is properly folded as deduced from CD spectra, and the immunodominant allergenic epitopes of the natural allergen are preserved in rPla l 1. These results allow to conclude that P.pastoris is a convenient system for the efficient production of biologically active rPla l 1, which could have a potential use for clinical purposes. Furthermore, a sequence similarity of Pla l 1 with the major allergen from the olive tree pollen, Ole e 1, is disclosed in this work, and the allergenic cross-reactivity between both allergens has been studied.

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[28] - Asero R, Mistrello G, Roncarolo D, Casarini M. Detection of allergens in plantain (Plantago lanceolata) pollen. Allergy 2000;55:1059-1062

BACKGROUND: Allergens in Plantago lanceolata have not been characterized yet. The objective was to characterize some plantain-pollen allergens and to investigate the cross-reactivity between plantain and grass pollens. METHODS: Sera from four patients monosensitive to plantain pollen and from eight grass-pollen-allergic patients showing strong skin reactivity to plantain pollen in the skin prick test (SPT) underwent immunoblot analysis with both Plantago and grass mix extract. Moreover, immunoblot inhibition experiments were done with grass mix extract as inhibitor. RESULTS: All four sera from plantain-allergic patients reacted to two distinct bands at 17 and 19 kDa, and 2/4 sera showed further reactivity to a 40-kDa protein, which in one case represented the most prominent IgE-binding allergen. Plantain-monosensitive subjects did not show any reactivity to grass-pollen extract, and preabsorption of their sera with grass-pollen extract did not cause any loss of reactivity to plantain pollen. Sera from all eight grass-pollen-allergic controls reacted to a 30-kDa protein in plantain pollen, and some sera showed cross-reactivity to higher and lower molecular-weight structures as well. In all cases, plantain reactivity was totally abolished by preabsorption of sera with grass-pollen extract. A preliminary investigation by immunoblot showed that polyclonal IgG anti-Phl p 5 (but not polyclonal Phl p 1) from rabbit reacted to a 30-kDa protein in plantain pollen. CONCLUSIONS: Three specific allergens (of 17, 19, and 40 kDa, respectively) have been detected in plantain pollen. Further studies on a larger number of patients will determine whether these proteins may be considered major allergens. Cross-reactivity between grass and plantain pollen is mainly caused by a 30-kDa protein in plantain pollen. Group 5 grass-pollen allergen is probably responsible for most grass/plantain cross-reactivity.

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[29] - Matthews PA, Baldo BA, Howden ME. Cytochrome c allergens isolated from the pollens of the dicotyledons English plantain (Plantago lanceolata) and Paterson's curse (Echium plantagineum). Mol Immunol 1988;25:63-68

Two cytochrome c allergens were isolated from extracts of the pollens of the dicotyledons English plantain (Plantago lanceolata) and Paterson's Curse (Echium plantagineum) by ion exchange chromatography, gel filtration and preparative isoelectric focusing. They were characterized by their absorption spectra, mol. wt, pI and amino acid composition. The cytochromes c bound specific IgE in the sera of hypersensitive patients by RAST. Preliminary evidence for allergenic cross-reactivity between them was obtained by RAST inhibition.

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[30] - 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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[31] - Grote M, Westritschnig K, Valenta R. Immunogold Electron Microscopic Localization of the 2 EF-Hand Calcium-Binding Pollen Allergen Phl p 7 and its Homologues in Pollens of Grasses, Weeds and Trees. Int Arch Allergy Immunol 2008;146:113-121

BACKGROUND: The 2 EF-hand calcium-binding allergen from timothy grass pollen, Phl p 7, contains the majority of relevant IgE epitopes among calcium-binding allergens occurring in pollen species of different plants . OBJECTIVE: To describe the ultrastructural localization of Phl p 7 allergen in timothy grass pollen and its homologues in a broad spectrum of allergologically relevant pollens from grasses (timothy grass, rye grass), trees (birch, alder, olive) and weeds (mugwort, ribwort, ragweed) commonly growing in Europe. MATERIALS AND METHODS: Mature pollens from 8 different plant species were collected and anhydrously prepared for transmission electron microscopy. In ultrathin sections, allergens were localized using an antibody prepared against a Phl p 7-derived peptide comprising the C-terminal half of the Phl p 7 wild-type molecule in combination with a secondary antibody coupled to 10-nm colloidal gold particles . RESULTS: Phl p 7 and Phl p 7 homologues were detected in pollen from each of the 8 pollen species investigated. The allergens were found in the cytoplasm of the pollen grains (cytoplasmic matrix, mitochondria, nuclei) and in the pollen wall (preferably the exine). Reserve materials were unlabeled . CONCLUSIONS: The 2 EF-hand calcium-binding allergen Phl p 7 from timothy grass and its homologues can be localized in all pollen species under investigation. This finding confirms that Phl p 7 is a marker allergen for sensitization of patients to a novel family of 2 EF-hand calcium-binding pollen allergens occurring in a number of important allergenic plants in Europe.

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[32] - Barletta B, Tinghino R, Iacovacci P, Afferni C, Butteroni C, Di Felice G, et al. Profilins, Ca-binding proteins, and CCD in 23 different pollen species. J Allergy Clin Immunol 2000;105(1 part 2):S331

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

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

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[34] - Tordesillas L, Sanchez-Monge R, Cuesta-Herranz J, Compes E, Garcia-Carrasco B, Lombardero M, et al. A search for peach Pru P 3-homologous lipid transfer proteins in pollens. Allergy 2008;63(suppl. 88):117-118

Background: Two different patterns of sensitisation to Rosaceae fruits have been described in the Centre/North (Bet v 1-like as main allergens) and South (Lipid Transfer Proteins, LTPs, as major allergens) of Europe. Sensitisation to distinctive pollens have been claimed as a putative source of these differential profiles. Objetive: To explore the presence of putative Pru p 3-like allergens in Ambrosia artemissifolia (ragweed), Artemisia vulgaris (mugwort), Betula verrucosa (birch), Dactylis glomerata (barnyard grass), Helianthus annus (sunflower), Parietaria judaica (pellitory), Phleum pratensis (timothy), Plantago lanceolata (plantain) and Platanus occidentalis (plane tree) pollens. Methods: PBS extracts from the different pollens were separated by SDS-PAGE and immunodetected with anti-Pru p 3 polyclonal antibodies or IgE of sera from peach allergic patients (with associated pollinosis). ELISA-inhibition assays using purified Pru p3 and mugwort Art v 3 allergens as inhibitors were also performed. Pollen RNA samples and a cDNA encoding Pru p 3 probe were used in Northern analysis. Results: Pru p 3-like proteins were detected only in mugwort, but not in the other pollens analyzed, using either anti-Pru p 3 antibodies or sera from allergic patients. The cDNA-Pru p 3 probe hybridized with specific RNA bands from mugwort and plane tree pollens, but no band was found in the rest of pollens. Conclusion: Pru p 3-like LTPs were present in mugwort and plane tree pollens, as previously reported, but were not detected in ragweed, mugwort, birch, barnyard grass, sunflower, pellitory, timothy, plantain and plane tree pollens. A limited structural similarity with Pru p 3 and or even null levels of putative homologous allergens could explain the lack of detection of Pru p 3-like LTPs in most pollen analyzed.

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[35] - Weber RW. Patterns of pollen cross-allergenicity. J Allergy Clin Immunol 2003;112:229-239

Knowledge of patterns of pollen cross-reactivity is crucial for diagnostics and especially for formulation of immunotherapy vaccines in times of diminishing availability of pollen extract constituents. As phylogenetic relationships have become better clarified, it becomes apparent that cross-reactivity does reflect taxonomy in the very great majority of cases. Contradictory observations of unexpected cross-reactivity between unrelated plants, sometimes remarkably distant ones, require explanation. There are many proteins, presumably performing vital functions, that are tightly preserved throughout the evolutionary tree from plants to animals, such as profilins, lipid transfer proteins, and pathogenesis-related proteins. These might function as panallergens. The small differences that exist between these ubiquitous proteins explain why these are frequently minor allergens not reacting in the majority of allergic sera. This review summarizes cross-reactivity studies with both crude pollen extracts and purified or recombinant allergenic proteins. The patterns of cross-allergenicity that emerge should be helpful in guiding both diagnostic and therapeutic decisions.

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[36] - Matthews PA, Baldo BA, Howden ME. Cytochrome c allergens isolated from the pollens of the dicotyledons English plantain (Plantago lanceolata) and Paterson's curse (Echium plantagineum). Mol Immunol 1988;25:63-68

Two cytochrome c allergens were isolated from extracts of the pollens of the dicotyledons English plantain (Plantago lanceolata) and Paterson's Curse (Echium plantagineum) by ion exchange chromatography, gel filtration and preparative isoelectric focusing. They were characterized by their absorption spectra, mol. wt, pI and amino acid composition. The cytochromes c bound specific IgE in the sera of hypersensitive patients by RAST. Preliminary evidence for allergenic cross-reactivity between them was obtained by RAST inhibition.

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[37] - Pham NH, Baldo BA, Bass DJ. Cypress pollen allergy. Identification of allergens and crossreactivity between divergent species. Clin Exp Allergy 1994;24:558-565

Studies employing sera from 34 subjects allergic to white cypress pine (Callitris glaucophylla) pollen identified 18 IgE antibody-binding components in the pollen of this species, five of which (MWs approximately 94, 68, 64, 43 and 34 kDa) were recognized by all of the sera. Protein blotting and quantitative inhibition studies revealed clear cross-reactivity between C. glaucophylla and Cupressus sempervirens pollen proteins and striking similarities in the IgE recognition band patterns of the two pollens. Inhibition experiments with other pollen extracts revealed that sera from C. glaucophylla pollen-allergic subjects can be divided into two groups--those inhibited only by extracts from the two Cupressaceae pollens and those inhibited both by these pollen proteins and by pollen extracts from other species. Most of the crossreactions in the latter group cannot be explained on the basis of taxonomic relationships or separate sensitizations. As with previous studies on birch and olive pollens, we conclude that pollen allergenic crossreactivity is much more wide-ranging than generally believed.

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[38] - 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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[39] - 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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[40] - Asero R, Mistrello G, Roncarolo D, Casarini M. Detection of allergens in plantain (Plantago lanceolata) pollen. Allergy 2000;55:1059-1062

BACKGROUND: Allergens in Plantago lanceolata have not been characterized yet. The objective was to characterize some plantain-pollen allergens and to investigate the cross-reactivity between plantain and grass pollens. METHODS: Sera from four patients monosensitive to plantain pollen and from eight grass-pollen-allergic patients showing strong skin reactivity to plantain pollen in the skin prick test (SPT) underwent immunoblot analysis with both Plantago and grass mix extract. Moreover, immunoblot inhibition experiments were done with grass mix extract as inhibitor. RESULTS: All four sera from plantain-allergic patients reacted to two distinct bands at 17 and 19 kDa, and 2/4 sera showed further reactivity to a 40-kDa protein, which in one case represented the most prominent IgE-binding allergen. Plantain-monosensitive subjects did not show any reactivity to grass-pollen extract, and preabsorption of their sera with grass-pollen extract did not cause any loss of reactivity to plantain pollen. Sera from all eight grass-pollen-allergic controls reacted to a 30-kDa protein in plantain pollen, and some sera showed cross-reactivity to higher and lower molecular-weight structures as well. In all cases, plantain reactivity was totally abolished by preabsorption of sera with grass-pollen extract. A preliminary investigation by immunoblot showed that polyclonal IgG anti-Phl p 5 (but not polyclonal Phl p 1) from rabbit reacted to a 30-kDa protein in plantain pollen. CONCLUSIONS: Three specific allergens (of 17, 19, and 40 kDa, respectively) have been detected in plantain pollen. Further studies on a larger number of patients will determine whether these proteins may be considered major allergens. Cross-reactivity between grass and plantain pollen is mainly caused by a 30-kDa protein in plantain pollen. Group 5 grass-pollen allergen is probably responsible for most grass/plantain cross-reactivity.

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[41] - Calabozo B, Diaz-Perales A, Salcedo G, Barber D, Polo F. Cloning and expression of biologically active Plantago lanceolata pollen allergen Pla l 1 in the yeast Pichia pastoris. Biochem J 2003;372:889-896

The glycoprotein Pla l 1 is the major allergen from Plantago lanceolata (English plantain) pollen, which is a common cause of pollinosis in temperate areas. Three complete cDNAs for Pla l 1 isoforms were isolated by polymerase chain reactions using specific 3' and 5' primers. All three Pla l 1 cDNAs code for a 25-residue leader peptide and a 131-residue mature protein that contains two polymorphic positions, an N-glycosylation site at position 107, and six cysteine residues involved in three disulfide bridges. The allergen variant Pla l 1.0101 was produced in P.pastoris at a yield of 20 mg per litre of culture as a mixture of non-glycosylated (17 kDa), glycosylated (23 kDa) and dimeric forms (32-39 kDa). rPla l 1 was purified by affinity chromatography with an anti-nPla l 1 monoclonal antibody, and its molecular and immunological properties were compared with the natural allergen by circular dichroism (CD) analysis, enzymatic deglycosylation, lectin binding assay, immunodetection and ELISA-inhibition assays using sera from plantain-allergic patients. The recombinant allergen is properly folded as deduced from CD spectra, and the immunodominant allergenic epitopes of the natural allergen are preserved in rPla l 1. These results allow to conclude that P.pastoris is a convenient system for the efficient production of biologically active rPla l 1, which could have a potential use for clinical purposes. Furthermore, a sequence similarity of Pla l 1 with the major allergen from the olive tree pollen, Ole e 1, is disclosed in this work, and the allergenic cross-reactivity between both allergens has been studied.

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[42] - Castro AJ, Alche JD, Calabozo B, Rodriguez-Garcia MI, Polo F. Pla l 1 and Ole e 1 pollen allergens share common epitopes and similar ultrastructural localization. J Investig Allergol Clin Immunol 2007;17(suppl. 1):41-47

BACKGROUND: English plantain (Plantago lanceolata L.) and olive (Olea europaea L.) pollens are important causes of pollinosis in large areas of North America, Australia, and the Mediterranean basin. The major pollen allergens of both plants, Pla I 1 and Ole e 1, share 38.7% of their amino acid sequences. OBJECTIVE: To analyze putative cross-reactivity between these 2 proteins. METHODS: Several antibodies and patients' sera were used in immunoblot and immunocytochemistry experiments. RESULTS: Two anti-Pla I 1 antibodies were able to bind to 3 polypeptides from olive pollen protein extracts, which correspond to the 3 glycosylation isoforms of Ole e 1 (18-22 kDa) previously described. Moreover, Pla I 1 protein was found in the cytoplasm of both the vegetative and the generative cells of P lanceolata mature pollen. On olive pollen sections, these anti-Pla I 1 antibodies displayed significant labeling in the cytoplasm of the vegetative cell and in both the exine and the material adhering to this outer layer of the pollen wall. In addition, the anti-Ole e 1 antibody 10H1 was found to cross-react with proteins of similar masses (16-20 kDa) to Pla I 1 variants. In Plantago pollen sections, the 10H1 antibody recognized proteins located in the cytoplasm of both the vegetative and generative cells. Cross-reaction was confirmed using sera from patients allergic to either plant pollen. CONCLUSION: Both allergens share common epitopes, which can be cross-recognized by different antibodies and sera from different patients, although this antigenic similarity seems to have little clinical relevance

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[43] - Aleman AM, Quirce S, Bombin C, Sastre J. Asma relacionada con la inhalacion de Plantago ovata. Med Clin (Barc) 2001;116:20-22

Psyllium is the seed of the plant Plantago ovata, which is widely used in nonprescription laxatives. It has been reported that psyllium powder may act as an inhalant allergen, especially in the workplace. A patient with asthma due to inhalation of P. ovata seed powder is described, being highlighted the allergenic potential of this substance frequently used as a laxative. We report a 31 year-old atopic woman, who handled and prepared at home, twice daily, a laxative containing P. ovata seeds (Plantaben) prescribed to her paralytic mother. METHODS AND RESULTS: Skin prick test to psyllium powder extract was positive. Specific IgE antibodies to psyllium were positive as determined by ELISA. Methacholine inhalation test revealed mild bronchial hyperresponsiveness (PC20 = 1.5 mg/ml). Specific bronchial challenge to psyllium powder elicited an isolated early asthmatic response. The allergens in psyllium were investigated by SDS-PAGE and immunoblotting, being identified several allergenic components with an apparent molecular weight of 66, 60, 36-20 and 14 kDa. No cross-reactivity was found between P. ovata seed and P. lanceolata pollen, as determined by ELISA-inhibition. CONCLUSION: Psyllium may act as a potent inhalant allergen capable of eliciting asthma symptoms, not only in an occupational context, but also in a domestic environment, affecting consumers of this laxative or relatives who handle it.

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[44] - Gauss WF, Alarie JP, Karol MH. Workplace allergenicity of a psyllium-containing bulk laxative. Allergy 1985;40:73-76

IgE antibodies specific to psyllium were demonstrated by RAST in a patient with work-place-related asthmatic and dermatological reactions. Antibodies were not cross-reactive with the related plant, English plantain.

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[45] - Rodriguez-Plata E, Poza-Guedes P, Gonzalez-Perez R, Hernandez-Santana G, Perez-Rodriguez E, Martinez-Tadeo J, et al. Sensitisation to Plantago ovata (Isphagula) and mustard: cross-reactivity versus co-sensitisation. Allergy 2008;63(suppl. 88):259

Background: The seed of the plant Plantago ovata is widely used in medicine by its laxative properties. It has been reported that this seed powder may act as an inhalant allergen, especially in the workplace. In several cases, has been asociated with food allergy.We report a 43 years old non atopic woman diagnosed of a moderate persistent intrinsic asthma and rhitinis who consulted by acute exacerbations of her respiratory symptoms when she handled at her work (nurse) a laxative containing Plantago ovata seeds. No cutaneous symptoms were associated. Besides, during the last year she had presented two episodes of urticaria and respiratory symptoms after eating an hamburger (the first time) and a hot dog (the second one). All vegetables and meats implicated were now tolerated. Methods: Skin prick tests were performed with a panel of common aeroallergens (including Plantago lanceolata), Plantago ovata and Mustard (comercial extract). Spirometry and rinometric studies were also performed. Specific IgE sensitisation were analized by CAP system (Pharmaciar) to several aeroallergens like pollens (Dactilys glomerata, Artemisia vulgaris, Parietaria officinalis, Plantago lanceolata) or dust mites (Dermatophagoides pteronyssimus and Dermatophagoides farinae), seeds of Plantago ovata, and Mustard. Laboratory: crossreactivity studies between Plantago ovata and Mustard were performed. Results: In vivo studies: Skin prick test were positive for Plantago ovata and Mustard. Negative lectures were obtained for all the other common aeroallergens. Spirometry: FVC 74%, FEV1 70%, FEV1/FVC 94% (in stable conditions). Rinometry: moderate bilateral obstruction. In vitro studies: total IgE was 60.18 UI/mL. Specific IgE values were: 6.92 kUI/L for mustard and 5.65 kUI/ L for Plantago ovata. Specific IgE were o 0.35 for all the aeroallergens tested. No immunoallergic cross-reactivity was found by immunoblot analysis. Conclusions: We describe a case of a patient with no previous history of atopic disease, who present specific sensitisation to Platago ovata in the workplace without sensitisation to other related pollens.Although mustard allergy was confirmed too, no immunologic cross-reactivity has been found between them.

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[46] - Bernedo N, García M, Gastaminza G, Fernández E, Bartolomé B, Algorta J, et al. Allergy to laxative compound (Plantago ovata seed) among health care professionals. J Investig Allergol Clin Immunol 2008;18:181-189

BACKGROUND: The seeds of Plantago ovata (psyllium, ispaghula) used in the manufacture of bulk laxatives are known to be the cause of occupational allergy (rhinitis, asthma) in health care and pharmaceutical workers. OBJECTIVE: We studied the prevalence of P ovata seed allergy among health care workers in geriatric care homes and compared it with a group of health care professionals not exposed to P ovata seed. Cross reactivity with Plantago lanceolata pollen was also studied. METHODS: Two groups of health professionals were recruited: 58 health care workers from geriatric care homes who were exposed daily to laxatives containing P ovata and 63 nonexposed health care professionals. The prevalence of allergy and sensitization to P ovata seed was determined based on clinical history, skin prick test, and analysis of specific immunoglobulin (Ig) E. IgE immunoblotting was performed to calculate the molecular weights of the P ovata seed allergens. Cross reactivity to P lanceolata pollen was studied by enzyme allergosorbent test (EAST) and immunoblot inhibition techniques. RESULTS: The prevalence of sensitization and clinical allergy to P ovata seed in the exposed group was 13.8% and 8.6%, respectively. No sensitization was observed in the nonexposed group. IgE-binding proteins of 17, 20, 25, 32-34, 54, 73-77, and > 97 kDa were identified. EAST inhibition and immunoblot inhibition demonstrated the existence of cross reactivity between P ovata seed and P lanceolata pollen extracts. CONCLUSIONS: The rate of sensitization to P ovata seed is high among health care workers in geriatric care homes (13.8%). A mild cross reactivity between P ovata seed and P lanceolata pollen was observed.

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[47] - Khalili B, Bardana EJ Jr, Yunginger JW. Psyllium-associated anaphylaxis and death: a case report and review of the literature. Ann Allergy Asthma Immunol 2003;91:579-584

BACKGROUND: Psyllium use has increased significantly in the United States in part due to its lipid-lowering property. The increased prevalence of consumption has led to its recognition as an emerging food allergen. OBJECTIVES: To report the case of a 42-year-old woman who experienced fatal anaphylaxis after ingesting a psyllium-based product and to review the literature. METHODS: The MEDLINE database was searched for articles from 1966 to 2002 using the keywords psyllium or ispaghula and each of the following: allergy, hypersensitivity, anaphylaxis, and asthma. Both English and non-English articles were included. RESULTS: Psyllium hypersensitivity has been well described in health care workers and pharmaceutical plant employees. Clinical manifestations of allergy range from upper respiratory tract symptoms on inhalation to anaphylaxis on ingestion. The prevalence of sensitization varies between these 2 groups. The allergenic epitope is not known. CONCLUSIONS: We present a case of psyllium hypersensitivity that resulted in death. There is a clear association between atopy and psyllium allergy. The case underscores the fact that even nonprescription "natural" products can be harmful to people with allergies.

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[48] - Arlian LG, Vyszenski-Moher DL, Lawrence AT, Schrotel KR, Ritz HL. Antigenic and allergenic analysis of psyllium seed components. J Allergy Clin Immunol 1992;89:866-876

The outer portions (husk) of psyllium seeds are a concentrated source of natural fiber used in some bulk-fiber laxatives and cereals. They are known to elicit respiratory allergic reactions after inhalation or ingestion among sensitized individuals. Antigenic and allergenic characterization of three psyllium-seed fractions (husk, endosperm, and embryo) was conducted with crossed immunoelectrophoresis (CIE), crossed radioimmunoelectrophoresis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to determine the source of psyllium allergenicity. Homologous CIE demonstrated psyllium endosperm and embryo extracts contained seven and four antigens, respectively. Husk extracts were too gelatinous to react by CIE. However, heterologous CIE profiles of endosperm or embryo extracts, reacted with antihusk antibodies, resulted in antigen-antibody precipitin peaks that matched the heavy staining precipitin lines of homologous reactions for endosperm and embryo, respectively. These results indicated that commercial-grade husk, endosperm, and embryo contained similar antigens. Extracts of all three seed components contained antigens that bound IgE antibodies in the sera of 11 psyllium RAST-positive individuals, as determined by crossed radioimmunoelectrophoresis. The few prominent husk protein/peptide bands resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were common in either embryo or endosperm. Immunoblots revealed common IgE reactive bands in all three seed fractions. Microscopic examination of the powdered commercial-grade psyllium (95% pure) revealed it contained endosperm and embryo particles. These immunologic, biochemical, and microscopic findings suggest that other contaminating seed components are primarily responsible for the allergenicity of commercial-grade psyllium-husk powder rather than the husk itself.

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[49] - Bernedo N, García M, Gastaminza G, Fernández E, Bartolomé B, Algorta J, et al. Allergy to laxative compound (Plantago ovata seed) among health care professionals. J Investig Allergol Clin Immunol 2008;18:181-189

BACKGROUND: The seeds of Plantago ovata (psyllium, ispaghula) used in the manufacture of bulk laxatives are known to be the cause of occupational allergy (rhinitis, asthma) in health care and pharmaceutical workers. OBJECTIVE: We studied the prevalence of P ovata seed allergy among health care workers in geriatric care homes and compared it with a group of health care professionals not exposed to P ovata seed. Cross reactivity with Plantago lanceolata pollen was also studied. METHODS: Two groups of health professionals were recruited: 58 health care workers from geriatric care homes who were exposed daily to laxatives containing P ovata and 63 nonexposed health care professionals. The prevalence of allergy and sensitization to P ovata seed was determined based on clinical history, skin prick test, and analysis of specific immunoglobulin (Ig) E. IgE immunoblotting was performed to calculate the molecular weights of the P ovata seed allergens. Cross reactivity to P lanceolata pollen was studied by enzyme allergosorbent test (EAST) and immunoblot inhibition techniques. RESULTS: The prevalence of sensitization and clinical allergy to P ovata seed in the exposed group was 13.8% and 8.6%, respectively. No sensitization was observed in the nonexposed group. IgE-binding proteins of 17, 20, 25, 32-34, 54, 73-77, and > 97 kDa were identified. EAST inhibition and immunoblot inhibition demonstrated the existence of cross reactivity between P ovata seed and P lanceolata pollen extracts. CONCLUSIONS: The rate of sensitization to P ovata seed is high among health care workers in geriatric care homes (13.8%). A mild cross reactivity between P ovata seed and P lanceolata pollen was observed.

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[50] - Morgan MS, Arlian LG, Vyszenski-Moher DL, Deyo J, Kawabata T, Fernández-Caldas E. English plantain and psyllium: lack of cross-allergenicity by crossed immunoelectrophoresis. Ann Allergy Asthma Immunol 1995;75:351-359

English plantain (Plantago lanceolata) weed pollen and psyllium (Plantago ovata) husk dust are inhalant allergens. Because of the phylogenetic relationship between these plant species, cross-allergenicity has been a concern. OBJECTIVE: The purpose of this study was to investigate the possible cross-allergenicity of plantain and psyllium. METHODS: Homologous and heterologous crossed immunoelectrophoresis (CIE) were performed using a commercial English plantain pollen extract and an extract of psyllium seed embryo. Crossed radioimmunoelectrophoresis (CRIE) was performed using sera from subjects who were RAST positive only to plantain (group A), RAST positive only to psyllium (group B), RAST positive to both plantain and psyllium (group C), or RAST negative to both (group D). RESULTS: All of the group A plantain subjects showed IgE binding to at least one of the six plantain allergens in homologous plantain CRIEs while only one of the sera from the group B subjects reacted very weakly to these plantain allergens. In homologous psyllium CRIE, all group B subjects showed pronounced IgE binding to 2 to 7 of the seven psyllium allergens. Several of the plantain subjects demonstrated only very weak binding to psyllium allergens. Heterologous CRIEs demonstrated little relevant IgE binding. CONCLUSIONS: These results indicate that there is little cross-allergenicity between psyllium husk and English plantain pollen.

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[51] - Aleman AM, Quirce S, Bombin C, Sastre J. Asma relacionada con la inhalacion de Plantago ovata. Med Clin (Barc) 2001;116:20-22

Psyllium is the seed of the plant Plantago ovata, which is widely used in nonprescription laxatives. It has been reported that psyllium powder may act as an inhalant allergen, especially in the workplace. A patient with asthma due to inhalation of P. ovata seed powder is described, being highlighted the allergenic potential of this substance frequently used as a laxative. We report a 31 year-old atopic woman, who handled and prepared at home, twice daily, a laxative containing P. ovata seeds (Plantaben) prescribed to her paralytic mother. METHODS AND RESULTS: Skin prick test to psyllium powder extract was positive. Specific IgE antibodies to psyllium were positive as determined by ELISA. Methacholine inhalation test revealed mild bronchial hyperresponsiveness (PC20 = 1.5 mg/ml). Specific bronchial challenge to psyllium powder elicited an isolated early asthmatic response. The allergens in psyllium were investigated by SDS-PAGE and immunoblotting, being identified several allergenic components with an apparent molecular weight of 66, 60, 36-20 and 14 kDa. No cross-reactivity was found between P. ovata seed and P. lanceolata pollen, as determined by ELISA-inhibition. CONCLUSION: Psyllium may act as a potent inhalant allergen capable of eliciting asthma symptoms, not only in an occupational context, but also in a domestic environment, affecting consumers of this laxative or relatives who handle it.

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[52] - Gauss WF, Alarie JP, Karol MH. Workplace allergenicity of a psyllium-containing bulk laxative. Allergy 1985;40:73-76

IgE antibodies specific to psyllium were demonstrated by RAST in a patient with work-place-related asthmatic and dermatological reactions. Antibodies were not cross-reactive with the related plant, English plantain.

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[53] - Rodriguez-Plata E, Poza-Guedes P, Gonzalez-Perez R, Hernandez-Santana G, Perez-Rodriguez E, Martinez-Tadeo J, et al. Sensitisation to Plantago ovata (Isphagula) and mustard: cross-reactivity versus co-sensitisation. Allergy 2008;63(suppl. 88):259

Background: The seed of the plant Plantago ovata is widely used in medicine by its laxative properties. It has been reported that this seed powder may act as an inhalant allergen, especially in the workplace. In several cases, has been asociated with food allergy.We report a 43 years old non atopic woman diagnosed of a moderate persistent intrinsic asthma and rhitinis who consulted by acute exacerbations of her respiratory symptoms when she handled at her work (nurse) a laxative containing Plantago ovata seeds. No cutaneous symptoms were associated. Besides, during the last year she had presented two episodes of urticaria and respiratory symptoms after eating an hamburger (the first time) and a hot dog (the second one). All vegetables and meats implicated were now tolerated. Methods: Skin prick tests were performed with a panel of common aeroallergens (including Plantago lanceolata), Plantago ovata and Mustard (comercial extract). Spirometry and rinometric studies were also performed. Specific IgE sensitisation were analized by CAP system (Pharmaciar) to several aeroallergens like pollens (Dactilys glomerata, Artemisia vulgaris, Parietaria officinalis, Plantago lanceolata) or dust mites (Dermatophagoides pteronyssimus and Dermatophagoides farinae), seeds of Plantago ovata, and Mustard. Laboratory: crossreactivity studies between Plantago ovata and Mustard were performed. Results: In vivo studies: Skin prick test were positive for Plantago ovata and Mustard. Negative lectures were obtained for all the other common aeroallergens. Spirometry: FVC 74%, FEV1 70%, FEV1/FVC 94% (in stable conditions). Rinometry: moderate bilateral obstruction. In vitro studies: total IgE was 60.18 UI/mL. Specific IgE values were: 6.92 kUI/L for mustard and 5.65 kUI/ L for Plantago ovata. Specific IgE were o 0.35 for all the aeroallergens tested. No immunoallergic cross-reactivity was found by immunoblot analysis. Conclusions: We describe a case of a patient with no previous history of atopic disease, who present specific sensitisation to Platago ovata in the workplace without sensitisation to other related pollens.Although mustard allergy was confirmed too, no immunologic cross-reactivity has been found between them.

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Les allergènes du plantain

Réactions croisées du plantain avec d’autres pollens

Réactions croisées du plantain avec d’autres produits

Le cas de l’ispaghule

Rubriques

Dans la même rubrique

Les Amaranthacées

Les ambroisies

L’armoise

Les Astéracées (Composées)

Les Euphorbiacées

Les Pariétaires