A monoclonal antibody specific for a carbohydrate epitope recognizes an IgE‐binding determinant shared by taxonomically unrelated allergenic pollens

Iacovacci, Patrizia; Pini, Carlo; Afferni, Claudia; Barletta, Bianca; Tinghino, Raffaella; Schininá, Eugenia; Federico, Rodolfo; Mari, Adriano; Felice, Gabriella Di

doi:10.1046/j.1365-2222.2001.01019.x

Cited by 36 publications

(33 citation statements)

References 33 publications

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“…These results are in accordance with the studies of Afferni et al [29] and Iacovacci et al [30], showing that a large portion of the IgE reactivity of Cupressaceae-allergic subjects is associated with sugar moieties of Cupa glycoproteins. Carbohydrate epitopes of Cupa pollen allergens were shown to be shared by taxonomically unrelated allergenic pollen species [31][32][33][34]. The N-linked oligosaccharide structures containing a (1-3) fucose and b(1-2) xylose are common features of many plant-derived glycoproteins.…”

Section: Discussionmentioning

confidence: 99%

Differential IgE sensitization to cypress pollen associated to a basic allergen of 14 kDa

et al. 2012

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The common cypress (Cupressus sempervirens) (Cups) pollen represents the first cause of respiratory allergies in the Mediterranean basin. The aim of this study was to characterize a novel 14-kDa cypress pollen allergen (BP14) allowing a clear dissociation of IgE sensitization patterns among allergic patients. The biochemical and immunochemical characterization of BP14 included determination of its isoelectric point, molecular mass, extraction kinetics, IgE binding prevalence, the presence of bromelain-type cross-reactive carbohydrate determinant and its IgE reactivity under reducing conditions. The presence of potential cross-reactive homologues in closely related cypress species, i.e. Cupressus arizonica (Cupa) and Cryptomeria japonica (Cryj), as well as in several taxonomically unrelated species was also investigated. According to our results, BP14 is easily and quickly solubilized in phosphate-buffered saline and exhibits several allergenic isoforms covering a broad range of pI (6.5-10.5). This allergen displays heat-stable conformational epitopes and does not include cross-reactive carbohydrate determinants, in contrast to high molecular weight cypress allergens. BP14 is expressed at higher levels in Cups than in Cupa and Cryj. No IgE cross-reactivity was found between the 14-kDa Cups pollen protein and proteins from some other non-Cupressaceae pollen allergenic sources such as orchard, timothy, wheat, maize, birch, ash and pine. Thus, IgE reactivity to BP14 is specific to Cupressaceae and discriminates two groups of patients allergic to cypress pollen. It might correspond to a relevant marker in relation to the sensitization process and ⁄ or the symptoms observed in some cypress-pollen-allergic patients. Furthermore, the description of BP14 should improve the diagnosis of cypress pollinosis.Abbreviations AB, ammonium bicarbonate buffer; BP14, basic protein of 14 kDa relative mass; CCD, cross-reactive carbohydrate determinant; Cryj,

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Section: Discussionmentioning

confidence: 99%

Differential IgE sensitization to cypress pollen associated to a basic allergen of 14 kDa

et al. 2012

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“…The use of monoclonal antibodies reactive to carbohydrate moiety allowed the detection of carbohydrate determinants in extracts from unrelated sources [17, 20, 21], further documenting the widespread reactivity of carbohydrate epitopes. Starting from a preliminary description of a wide CCD-IgE binding to clinically irrelevant allergenic extracts [6], we tried to define the degree of interference of CCD-IgE in the IgE detection.…”

Section: Discussionmentioning

confidence: 99%

“…In fact, in the present study, 21% of the patients having CCD-IgE displayed a cutaneous reactivity to the purified HRP. None of these patients reacted to the other tested glycoproteins having a different number of glycan side chains and also different glycan structures [2], though such molecules have been shown to be CCD-IgE reactive [6, 21]. The heterogeneous distribution among the subsets of respiratory patients (e.g.…”

Section: Discussionmentioning

confidence: 99%

“…From an allergological perspective, the most important feature of CCD is the wide distribution among plant-derived proteins [17], though similar structures can also be present in non-plant glycoproteins [1, 15]. Monoclonal and polyclonal antibodies to CCD have been used to define reactive carbohydrate epitope distribution [10, 17, 18, 19, 20, 21], whereas limited data are available on the prevalence and distribution of IgE reactive with CCD [6, 15, 22, 23]. …”

Section: Introductionmentioning

confidence: 99%

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IgE to Cross-Reactive Carbohydrate Determinants: Analysis of the Distribution and Appraisal of the in vivo and in vitro Reactivity

Mari¹

2002

Int Arch Allergy Immunol

186

210

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Background: IgE to cross-reacting carbohydrate determinants has already been described by several authors, but their function and distribution are still a matter of debate. In previous studies we showed how the presence of IgE to bromelain could be a useful and simple marker of the presence of IgE to carbohydrate epitopes. Methods: A survey of 1,831 subjects with a suspected allergic respiratory disease has been carried out by detecting IgE to bromelain. Data were analysed on the basis of demographical and allergological parameters. To find out whether a glycoprotein is capable of triggering an allergic reaction, 1,076 subjects were also skin tested with several purified molecules bearing carbohydrate side chains differing in number, composition and complexity. Results: An overall prevalence of 23% of positive IgE to cross-reacting carbohydrate determinants was recorded. Prevalence varied when subsets of non-allergic (5%), non-pollen-allergic (10%), and pollen-allergic (31%) subjects were considered. Prevalence further increased in subsets with multiple pollen sensitization (71%), and with a previous pollen immunotherapy course (46%), whereas minor differences were found in gender and age distribution. Almost all the allergenic extracts recorded negative in the skin test gave a positive IgE test in vitro. A higher correlation was found mainly with plant-derived allergenic extracts, whereas a lower one was recorded with mites and fungi. Horseradish peroxidase was the only glycoprotein capable of exerting a positive skin test in 21% of the subjects with IgE to cross-reacting carbohydrate determinants, 80% of them having IgE to the HRP molecule. Conclusions: IgE to cross-reacting carbohydrate determinants are common among the allergic population and the binding to skin test negative allergenic extracts further confirms their poor biological activity. Further studies on horseradish peroxidase should be carried out to define the role of the glycan side chains in its allergenic activity.

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“…ELISA and ELISA inhibition were carried out as previously described [22, 23]with minor modifications. Polystyrene plates (Dynatech, Plochingen, Germany) were coated with 0.2 ml/well of a solution containing 5 µg/ml of rOle e 5.…”

Section: Methodsmentioning

confidence: 99%

Cloning and Expression of the <i>Olea europaea</i> Allergen Ole e 5, the Pollen Cu/Zn Superoxide Dismutase

Butteroni

Afferni²,

Barletta³

et al. 2005

Int Arch Allergy Immunol

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Background: Recombinant DNA technology does provide pure, well-defined and reproducible products to be used for clinical purposes, by cloning and expressing the cDNA of allergens present in a specific extract. Ole e 5 is a pollen allergen of Olea europaea with an IgE-binding frequency of about 35%, which has been identified as a superoxide dismutase (SOD). The aim of this study was to clone the cDNA of Ole e 5, to express Ole e 5 in Escherichia coli and to characterize its immunoreactivity. Methods: cDNA of Ole e 5 was amplified by nested 3′-RACE PCR and cloned in pGEX vector 6P expression vector. After sequencing of some clones and homology analysis, the rOle e 5 was produced in an E. coli strain as a fusion protein with GST and purified. Then, the protein immunoreactivity was evaluated by patients’ IgE binding (ELISA, ELISA inhibition, and immunoblotting) and by rabbit anti-rOle e 5 binding (immunoblotting and immunoblotting inhibition). Results: The sequence analysis of Ole e 5 cDNA confirmed that Ole e 5 is a Cu/Zn SOD, with an identity from 90 to 80% with SOD from other species. rOle e 5 was recognized by IgE from 39% of olive pollen-allergic patients tested; moreover, this binding was inhibited by the olive pollen extract. An anti-rOle e 5 antiserum raised in rabbit strongly reacted with a natural component of about 16-kDa molecular weight present in the olive pollen extract; moreover, this binding was inhibited by the recombinant protein. Conclusions: Ole e 5 is the first Cu/Zn SOD identified as an allergen in a pollen source. Due to the widespread presence of this enzyme, rOle e 5 allergen, cloned and expressed in a complete form in E. coli, could represent a good tool to investigate the allergen cross-reactivity between O. europaea pollen and other allergenic sources, such as plant foods and other pollens.

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A monoclonal antibody specific for a carbohydrate epitope recognizes an IgE‐binding determinant shared by taxonomically unrelated allergenic pollens

Cited by 36 publications

References 33 publications

Differential IgE sensitization to cypress pollen associated to a basic allergen of 14 kDa

Differential IgE sensitization to cypress pollen associated to a basic allergen of 14 kDa

IgE to Cross-Reactive Carbohydrate Determinants: Analysis of the Distribution and Appraisal of the in vivo and in vitro Reactivity

Cloning and Expression of the <i>Olea europaea</i> Allergen Ole e 5, the Pollen Cu/Zn Superoxide Dismutase

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