Oleaceae pollen allergy and its cross-reactivity in the mediterranean area in the context of globalization.

Opening frontiers has led in the last two decades to a massive migration of Romanians to Spain and Italy, two countries known to have large areas planted with olives. Romania owns large areas planted with ash forests. Because of cross-reactivity between these two trees belonging to the Oleaceae family and the big allergenic potential of olive pollens, after a number of years of residence in these countries, many Romanians will present allergic respiratory symptoms of rhinitis and asthma, both in the season of olive pollination and ash pollination.

Towards a global vision of molecular allergology: a map of exposure to airborne molecular allergens.

Allergy diagnostics have changed in the last 10-15 y, moving from the use of extracts for in vivo and in vitro diagnosis to the Component Resolved Diagnosis, based on purified or recombinant allergens. As expected, aerobiology developed similarly, and measurement of allergens in both outdoor and indoor air is now feasible. With the aim of promoting a global view of molecular allergy, we have drawn a map of exposure to molecular aeroallergens in Italy on the bases of geo-climatic regions, maps of pollen distribution, and published data on the molecular profile of sensitization in Italian patients. Given the latitudinal extension of Italy, the profile of exposure to some allergens, such as those of the "Birch Group" and weeds, varies greatly from North to South, while the distribution of exposure to grass allergens is more homogeneous. This map can contribute to a global molecular vision of allergy, helping clinicians to view exposure to pollen in a new way. The exposure profile of the area where patients live can also indicate the correct choice of molecular diagnostics and, therefore, of the appropriate allergen immunotherapy.

Analysis of hypersensitivity to oleaceae pollen in an olive-free and ash-free area by commercial pollen extracts and recombinant allergens.

BACKGROUND: Olive pollen sensitization is surprisingly frequent in Milan, an area that is virtually both olive- and ash-free. OBJECTIVE: To establish the prevalence of olive pollen sensitization north of Milan, and to investigate the allergens involved. METHODS: 300 pollen-allergic patients living in this area were randomly selected. Based on SPT results, olive pollen reactors were classified as multi-sensitized, oligo-sensitized, or mono-sensitized. IgE to markers of primary sensitization to olive pollen (Ole e 1), as well as to pollen pan-allergens such as profilin (Phl p 12) or polcalcin (Phl p 7) were measured. Further, the putative cross-reactivity between grass group XI allergen and Ole e 1 was investigated. RESULTS: 107 (36%) patients were sensitized to olive pollen; 67 (63%) were multi-sensitized, while only 4 (4%) were mono-sensitized. Specific IgE to Ole e 1 were found in 32/46 (70%) cases; 22 of them (69%) co-recognized pollen pan-allergens, as shown by IgE reactivity to Phlp 7 and/or Phlp 12. Sera from 14 (30%)patients did not react to Ole e 1; of these, 10 (71%) were pan-allergens reactors. No correlation was found between IgE levels to Phl p 11 and Ole e 1. CONCLUSIONS: A majority of olive pollen-sensitized subjects seen in the surroundings of Milan are truly allergic to Oleaceae. In the absence of both olive and ash trees exposure to privet pollen might represent the source of this kind of sensitization.

[Sensitization to pollens of Oleaceae family in a group of patients from Mexico City]. / Sensibilización a pólenes de la familia Oleaceae en un grupo de pacientes de la Ciudad de México.

BACKGROUND: In Mexico City, where the Olive trees are scarce, the Oleaceae family has been recognized as one of the most important causes of pollen allergy, possibly related to cross-reactivity with other pollens of the Oleaceae family, as Fraxinus excelsior, Ligustrum vulgaris and Osmanthus americanus. OBJECTIVE: To know the prevalence of sensitization to the Oleaceae family in Mexico City, cross-reactivity, as well as concurrent sensitizations, and vegetables and fruits associated to oral allergy syndrome (OAS). PATIENTS AND METHOD: A prospective, longitudinal study was carried out during two months, among patients who attended the outpatient allergy clinic of the General Hospital of Mexico with symptoms suggestive of respiratory allergy. A questionnaire to search OAS was used, and skin Prick tests to pollens were performed. RESULTS: Among 279 patients, 164 (58.8%) had respiratory allergy and positive skin prick test (SPT) to pollens. Olea europaea had the highest prevalence of SPT reactivity (52.4%), followed by Fraxinus excelsior (43.3%), Ligustrum vulgaris (36.6%) and Osmanthus americanus (15.9%). There were OAS in 53.5%, 50.7%, 50% and 53.8%, respectively, mainly related to peach, apple, kiwi, pear and banana. Olive monosensitization was found in 5.5%. CONCLUSION: In this study, Olea europaea had the highest prevalence of positive reactivity to pollens in Mexico City, then its sensitization must be determined routinely.

Seasonal idiopathic rhinitis with local inflammatory response and specific IgE in absence of systemic response.

BACKGROUND: Patients with idiopathic rhinitis (IR) are considered to be nonallergic because they have a negative skin prick test (SPT) and allergen specific-IgE in serum. The concept of localized mucosal allergy in the absence of atopy has recently been proposed. The immunological mechanisms involved in seasonal IR have not been sufficiently studied. We examined nasal mucosa inflammation, the presence of nasal specific-IgE and the response to nasal allergen provocation test (NAPT) in patients with seasonal IR who presented symptoms only in spring. METHODS: We evaluated 32 patients with seasonal IR and 35 with persistent allergic rhinitis to pollen (PAR-P) and compared these with healthy controls and persons with PAR to house dust mite during the pollen season, as well as by NAPT out-of-season with grass and Olea europea. We measured the nasal leukocyte-lymphocyte phenotype (CD45, CD33, CD16, CD3, CD4 and CD8), eosinophil-cationic-protein, and total and specific-IgE to grass and olive pollen in serum and nasal lavage and performed NAPT. RESULTS: In the IR group, 62.5% had a positive NAPT (IR-PosNAPT), 20/32 to grass, with four of these having a positive NAPT to olive pollen as well. IR-PosNAPT patients showed a similar nasal leukocyte-lymphocyte profile to the PAR-P patients and different to controls. We detected nasal specific-IgE in 35% of IR-PosNAPT patients. CONCLUSIONS: These results support the hypothesis that a subgroup of patients with IR have seasonal symptoms with evidence of a nasal allergic immune reaction in the absence of a positive SPT or serum specific IgE.

Vegetable proteomics: the detection of Ole e 1 isoallergens by peptide matching of MALDI MS/MS spectra of underivatized and dansylated glycopeptides.

Ole e 1 (NCBI entry gi|14424429) is the major allergen of Oleaceae family. Multiple isoforms and variants are present in varying degrees of distribution. In this report, we present a new approach to the resolution of multiple forms of Ole e 1 from whole antigen extracts, based on a preliminary chemical fractionation procedure followed by MALDI MS and MS/MS measurements. The characterization of Ole e 1 isoallergens was accomplished through the identification of the amino acid sequence including the glycosylation site and the structure of the glycan moieties. The structure feature of the identified Ole e 1.0102 (gi|2465127), main olive allergen [Olea europaea] (gi|13195753), Major pollen allergen Ole e 1 (gi|33329740) and Ole e 1c (gi|1362131) is represented by the point mutation K(106) --> I and by the presence of a glycan moiety. Two other variants Major pollen allergen (Allergen Ole e1) (Ole e I) (gi|14424429) and Ole e 1.0103 protein [Olea europea] (gi|2465129) were identified as nonglycosylated species. These results, partially in disagreement with Swiss-Prot annotation, were validated by matching the MALDI MS/MS spectra of the natural tryptic mixture with those obtained after deglycosylation.

Associations among pollen sensitizations from different botanical species in patients living in the northern area of Madrid.

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

Hypoallergenic mutants of Ole e 1, the major olive pollen allergen, as candidates for allergy vaccines.

BACKGROUND: The C-terminal region of Ole e 1, a major allergen from olive pollen, is a dominant IgE-reactive site and offers a target for site-directed mutagenesis to produce variants with reduced IgE-binding capability. OBJECTIVE: To evaluate in vitro and in vivo the immunogenic properties of three engineered derivatives of Ole e 1. METHODS: One point (Y141A) and two deletion (135Delta10 and 140Delta5) mutants were generated by site-directed mutagenesis of Ole e 1-specific cDNA and produced in Pichia pastoris. Ole e 1 mutants were analysed for IgE reactivity by ELISA using sera from olive pollen-allergic patients. Their allergenicity was also investigated in both a mouse model of allergic sensitization and in basophil activation assays. IgG1 response was assayed by immunoblotting and competitive ELISA. T cell reactivity was evaluated by proliferation assays and cytokine production in splenocyte cultures. RESULTS: The 135Delta10 mutant showed the strongest reduction in the IgE-binding capability of sera from olive pollen-allergic patients. Rat basophil leukaemia assays identified the deletion mutant 135Delta10 as the variant with the lowest beta-hexosaminidase-releasing capacity. Furthermore, the same 135Delta10 mutant induced the lowest IgE levels in a BALB/c mouse model of sensitization. All Ole e 1 mutants retained their allergen-specific T cell reactivity. Immunization of mice with the mutants induced IgG1 antibodies, which cross-reacted with Ole e 1 and Ole e 1-like allergens from ash, lilac and privet pollens. The ability of the human IgE to block the binding of anti-Ole e 1 mutant-specific mouse IgG1 antibodies to natural Ole e 1 demonstrated that Ole e 1 mutants are able to induce in vivo antibodies reactive to the natural allergen. CONCLUSION: The 135Delta10 mutant with reduced allergenicity, intact T cell reactivity and capacity to induce blocking antibodies could provide a suitable candidate vaccine for efficient and safer therapy of olive pollen allergy.

The major allergen of olive pollen Ole e 1 is a diagnostic marker for sensitization to Oleaceae.

BACKGROUND: Trees of the family Oleaceae are important allergen sources, with a strongly varying geographic distribution. For example, olive pollen is an important allergen source in Mediterranean countries, whereas ash pollen dominates in Northern and Central Europe and North America. The aim of this study was to compare the profiles of olive and ash pollen allergens and to study the degree of cross-reactivity using populations of allergic patients selectively exposed to olive or ash pollen. METHODS: Olive and ash pollen extracts were analyzed by IgE immunoblotting using sera from Spanish patients highly exposed to olive pollen and Austrian patients without olive but ash pollen exposure. IgE cross-reactivity was studied by qualitative immunoblot inhibition assays and semiquantitative ELISA inhibitions using olive, ash, birch, mugwort, timothy grass pollen extracts and the major olive pollen allergen, Ole e 1. RESULTS: Spanish and Austrian patients exhibited an almost identical IgE-binding profile to olive and ash pollen allergens, with major reactivity directed against Ole e 1, and its homologous ash counterpart, Fra e 1. IgE inhibition experiments demonstrated extensive cross-reactivity between olive and ash pollen allergens. However, whereas cross-reactions between profilins and calcium-binding allergens also occurred between unrelated plant species, cross-reactivity to Ole e 1 was confined to plants belonging to the Oleaceae. CONCLUSIONS: Ole e 1 is a marker allergen for the diagnosis of olive and ash pollen allergy.

Bronchodilatory effects of the aqueous extract of Gynostemma pentaphyllum and gypenosides III and VIII in anaesthetized guinea-pigs.

The bronchodilatory activity of the aqueous extract of Gynostemma pentaphyllum Makino leaves was investigated in anaesthetized guinea-pigs and compared with two of its isolated gypenosides (III and VIII). The results showed that the intravenous administration of the decoction of G. pentaphyllum (2.5, 5 or 10 mg kg(-1)) decreased bronchial resistance in basal conditions and significantly (P < 0.01) reduced (68% inhibition) the bronchoconstrictor action of histamine. Furthermore, the extract antagonized (80% inhibition) the bronchoconstrictor response induced by the antigen in sensitized guinea-pigs. Gypenosides III (0.7 mg kg(-1), i.v.) and VIII (0.3 mg kg(-1), i.v.) caused a similar protective effect in both experimental models used; however, the duration and the intensity of the action was less than that of the extract containing corresponding quantities of gypenosides III and VIII. This study confirmed the validity of the traditional use of this plant in the treatment of asthma and other respiratory disorders.

Olive pollen allergen Ole e 8: identification in mature pollen and presence of Ole e 8-like proteins in different pollens.

In a first approach, Ole e 8, a novel Ca2+-binding protein from olive pollen, was cloned and produced in Escherichia coli. We have obtained the natural form of Ole e 8 (nOle e 8) from the pollen and examined its immunologic equivalence with its recombinant form (rOle e 8). Size exclusion chromatography and a phenyl-Sepharose CL-4B affinity column were used to obtain nOle e 8 from the olive pollen. Inhibition assays by immunoblotting, using rOle e 8-specific rabbit antiserum, were performed to analyze the immunologic equivalence between the natural and the recombinant allergen, as well as to detect its presence in other pollens. Recombinant and natural Ole e 8 resulted immunologically equivalents, since they completely inhibited the IgG binding of the polyclonal antiserum to each other. Ole e 8-like proteins were detected in Oleaceae and Juniperus communis pollen, and might contribute to cross-reactivity processes between taxonomically related pollens.

Melatonin and beta-endorphin changes in children sensitized to olive and grass pollen after treatment with specific immunotherapy.

BACKGROUND: Specific immunotherapy for respiratory allergy, a seasonal disease, significantly reduces the inflammatory process, attenuating the clinical symptoms. The mechanism for the clinical beneficial effect of immunotherapy has not yet been clarified. Melatonin shows a circadian and seasonal variation and together with the endogenous opioid system plays an immunomodulatory role acting on both specific and nonspecific immunity responses. Thus, the possibility that immunotherapy involved changes in the melatonin-opioid system was investigated. METHODS: Thirty-five children aged 3-15 years with rhinitis and asthma due to olive + grass pollen sensitization were studied. The patients were treated with depot extracts containing the identified allergens with increasing doses from 1 to 1,000 IU/ml during 3 months. Melatonin, beta-endorphin, total and specific IgE and IgG4 were determined before and after treatment. RESULTS: All children showed a significant improvement of their symptoms at the end of the treatment, coinciding with a significant drop of both melatonin and beta-endorphin levels. Total IgE decreased in most of the cases although the mean values did not show significant changes. Specific IgE and IgG4 were also unchanged. A significant correlation between melatonin and beta-endorphin and between beta-endorphin and IgG4 was found before immunotherapy, and these correlations disappeared when the treatment was finished. CONCLUSIONS: The decrease in the levels of melatonin and beta-endorphin suggests the disappearance of their stimulating influence on the immune system. Thus, hyposensitization after immunotherapy may involve, at least in part, the inhibition of the immunoenhancing properties of the melatonin-opioid system.

Preliminary assessment of the immune response to Olea europaea pollen extracts encapsulated into PLGA microspheres.

In this study, poly (D,L-lactide-co-glycolide) (PLGA) microspheres encapsulating Olea europaea pollen extracts were prepared by using the double emulsion (w/o/w) based on a solvent evaporation/extraction method. The resulting microspheres were 1.93 microns in size. The total allergen loading and surface-associated allergen were 8 and 0.64%, respectively. The release of the allergen from the microspheres showed a biphasic profile with an initial burst release followed by a sustained release phase. Finally, the polyacrylamide gel electrophoresis (SDS-PAGE) results showed that the encapsulation process does not affect the stability of the protein. We describe here some preliminary observations concerning the use of these microspheres as parenteral antigen delivery systems for immunization with O. europaea pollen extracts, in a small animal model, the mouse.