Mugwort Pollen Research Articles

Humoral and Cellular Cross-Reactivity between Amb a 1, the Major Ragweed Pollen Allergen, and Its Mugwort Homolog Art v 6

Ragweed and mugwort are closely related weeds that represent the major cause of pollen allergy in late summer. Concomitant sensitization and clinical cross-reactivity frequently occur in subjects who are coexposed to both pollen species, and have implications for diagnosis and specific immunotherapy. Molecules involved in this cross-reactivity might be Amb a 1, the major ragweed pollen allergen, and Art v 6, a highly homologous allergen from mugwort. Therefore, we investigated the IgE and T cell response to Art v 6 of 60 weed pollen-allergic patients and assessed its immunological cross-reactivity with Amb a 1. Results of ELISA inhibition experiments suggested that both allergens are largely cross-reactive, but Amb a 1 possesses more IgE epitopes than Art v 6. In patients with IgE to both allergens, Amb a 1-induced T cell lines and clones responded weakly to Art v 6. Moreover, Art v 6-induced T cell lines responded stronger to Amb a 1. T cell epitope mapping of Art v 6 revealed that it contains only a few cross-reactive epitopes, which is opposed to the multiple T cell-activating regions present in Amb a 1. In summary, Amb a 1 can elicit more diverse allergen-specific IgE and T cell responses than Art v 6 and dominates the cross-reactivity with its homolog. Nevertheless, Art v 6 can act as a primary sensitizing allergen in areas with high mugwort pollen exposure, and consequently may facilitate sensitization to Amb a 1 by epitope cross-recognition of T and B cells.

Oral administration of allergen extracts from mugwort pollen desensitizes specific allergen-induced allergy in mice

Clinically, sublingual immunotherapy (SLIT) using allergen extracts effectively alleviates the symptoms of allergic rhinitis and asthma. We hypothesized that oral administration of a high-dose of allergen extracts imitates SLIT, which may prevent IgE-related responses in allergic diseases. In the present study, we investigated the effects of oral administration of allergen extracts from mugwort pollen (MP) on allergen-induced inflammation and airway hyperresponsiveness (AHR) in an allergic mouse model. After administration of MPdrop containing Art v 1 and Art v 4 extracts derived from MP specifically in MP-sensitized mice, the effects of MPdrop on AHR, inflammatory cell accumulation, cytokine production in the bronchoalveolar lavage fluid and lung tissue, and serum IgE and IgG levels were investigated. The results indicated that MPdrop not only prevented the AHR in response to methacholine in a dose-dependent manner but also significantly reduced the total serum and allergen-specific IgE levels. All of the maximal effects were achieved at a dose of 100μg/(kgd) and were comparable to the effects of dexamethasone at a dose of 0.5mg/(kgd). Furthermore, oral administration of MPdrop dose-dependently elevated allergen-specific serum IgG2a levels, reduced total and allergen-specific IgE levels and normalized the imbalance between the Th1 cytokine IL-12 and Th2 cytokines IL-4 and IL-5. Finally, oral administration of MPdrop significantly reduced goblet cell hyperplasia and eosinophilia in the MP-sensitized allergic mouse model. These data suggest that MPdrop effectively improves specific allergen-induced inflammation and AHR in MP-sensitized and -challenged mice and provides the rationale for clinical use of MPdrop in the specific allergen-induced asthma.

Pollen concentrations of some plants in the air over Olszanica (Bieszczady Niskie Mountains) and Wrocław in the 2008 season

This paper presents the results of an analysis of pollen season patterns for taxa which show the strongest allergenic activity (alder, birch, grasses, and mugwort) in 2008 in the air over Wrocław and Olszanica. The study was carried out using the volumetric method (Burkard trap). The results show variation in pollen seasons between the analyzed localities. An attempt was made to find out in which of the sites in question - the urban site or the rural one - there was a greater risk of allergens of the selected plants. The results of the present study show that the alder, birch and grass pollen seasons in 2008 started and ended earlier in Wrocław, and maximum pollen concentrations were definitely lower. But the mugwort pollen season started earlier and ended much later in Olszanica, while maximum pollen concentration of this taxon was more than twice lower than in Wrocław. In 2008 in the investigated localities, the highest pollen concentrations of the plants in question occurred in the following months: alder in February, birch in April, grasses in June, while mugwort in August. In 2008 alder and birch pollen allergen risk was comparable in the investigated urban and rural environment. However, grass and mugwort pollen allergens posed a significantly greater threat in the rural environment than in Wrocław.

IgE Reactivity Patterns in Patients with Allergic Rhinoconjunctivitis to Ragweed and Mugwort Pollens

Differential diagnosis between ragweed and mugwort pollen allergy represents a large clinical problem in areas where both plants are present. The aim of this study was to investigate ragweed- and mugwort-sensitized patients to identify specific IgE reactivity profiles. Results were correlated to clinical findings such as medical history and health-related quality of life (HRQL). Seventy-four patients with allergic rhinoconjunctivitis between July and October were examined and underwent in vivo tests (skin-prick test [SPT] and nasal provocation). Sera were evaluated for IgE reactivity to mugwort and ragweed pollen extracts, major (Art v 1; Amb a 1) and minor (profilin and calcium-binding protein) allergens. HRQL was evaluated using a standardized questionnaire. Seventy-one patients revealed positive SPT reactivity against mugwort and 60 patients against ragweed extracts. Of these patients, 74 revealed IgE antibodies against mugwort extracts, whereas anti-Art v 1 antibodies were detectable in 50 individuals. Fifty-five patients showed IgE antibodies against natural ragweed extracts; anti-Amb v 1 antibodies were detected in six cases only. Using standardized clinical history and HRQL questionnaires we were not able to detect any differences within different reactivity patterns. Within the investigated population of 74 weed-allergic patients the prevalence of true mugwort and ragweed sensitization can be calculated as 68 and 8%. High prevalence of ragweed sensitization when testing with full extracts can be explained by cross-reactivity between other weeds, e.g., mugwort rather than cosensitization. Differences in medical history and HRQL between different reactivity patterns were not detectable.

Mugwort (Artemisia L.) pollen in aeroplankton of Lublin, 2001-2005

The course of the <i>Artemisia</i> pollen season was recorded in Lublin over a period of five years: 2001-2005. The volumetric method was applied in the studies, using a VPPS 2000 Lanzoni trap. The length of the season was determined by the 98% method. The impact of several meteorological factors on the start and course of the pollen season was analysed. It was found that in the five-year period studied the mugwort pollen season started in the second or third decade of July and lasted 59-90 days. Maximum concentrations in the range of 103-221 pollen grains in 1 m<sup>3</sup> of air were noted between 2 and 9 August. Annual totals of mugwort pollen grains ranged from 1496 to 2532. A significant positive correlation was demonstrated between the <i>Artemisia</i> pollen concentration and air temperature, and a negative correlation between the pollen concentration and air relative humidity and cloudiness. A significant impact of temperature on the start of the <i>Artemisia</i> pollen season was also found.

Meteorological factors and pollen season dynamics of selected herbaceous plants in Szczecin, 2004-2008

The pollen of mugwort, plantain, sorrel, nettle and pigweed is an important airborne allergen source worldwide. The occurrence of pollen grains in the air is a seasonal phenomenon and estimation of seasonal variability in the pollen count permits evaluation of the threat posed by allergens over a given area. The aim of the study was to analyse the dynamics of <i>Artemisia</i>, <i>Plantago</i>, <i>Rumex</i>, <i>Urticaceae</i> and Chenopodiaceae pollen season in Szczecin (western Poland) in 2004-2008 and to establish a relationship between the meteorological parameters versus the pollen count of the taxa studied. Measurements were performed by the Hirst volumetric trap (model Lanzoni VPPS 2000). Consecutive phases during the pollen season were defined for each taxon (1, 5, 25, 50, 75, 95, 99% of annual total) and duration of the season was determined using the 98% method. On the basis of this analysis, temporary differences in the dynamics of the seasons were most evident for <i>Artemisia</i>. Correlation analysis with weather parameters demonstrated that the maximum wind speed, mean and maximum air temperature, relative humidity and dew point are the main factors influencing the average daily pollen concentrations in the atmosphere.

Pollen production in selected species of anemophilous plants

In the study, structural features of flowers of the following allergenic plant species were analysed: <i>Betula verrucosa</i>, <i>Secale cereale</i>, <i>Rumex acetosella</i>, <i>Plantago major</i> and <i>Artemisia vulgaris</i>. Pollen production was established by calculating the number of pollen grains produced by the stamen, flower and inflorescence. The dates of occurrence and pollen grains concentration in the air of Lublin were determined. A positive correlation was found between the length of anthers and the number of pollen grains produced. The largest number of pollen grains per anther is produced by Secale cereale (22 360), whereas the smallest one by Plantago major (5 870). The other species produced intermediate numbers of pollen grains in the anther: <i>Betula verrucosa</i> - 11 160, <i>Rumex acetosella</i> - 10 850, <i>Artemisia vulgaris</i> - 9 580. The birch pollen season in Lublin lasts about a month, and pollen of this taxon reaches the highest airborne concentrations among the studied taxa. Low values of pollen concentrations are characteristic for rye and plantain, whereas slightly higher values are recorded for sorrel pollen. Mugwort pollen reaches high concentrations which are noted at the beginning of August.

Expression of the major mugwort pollen allergen Art v 1 in tobacco plants and cell cultures: problems and perspectives for allergen production in plants

An economic and cheap production of large amounts of recombinant allergenic proteins might become a prerequisite for the common use of microarray-based diagnostic allergy assays which allow a component-specific diagnosis. A molecular pharming strategy was applied to express the major allergen of Artemisia vulgaris pollen, Art v 1, in tobacco plants and tobacco cell cultures. The original Art v 1 with its endogenous signal peptide which directs Art v 1 to the secretory pathway, was expressed in transiently transformed tobacco leaves but was lost in stable transformed tobacco plants during the alternation of generations. Using a light-regulated promoter and “hiding” the recombinant Art v 1 in the ER succeeded in expression of Art v 1 over three generations of tobacco plants and in cell cultures generated from stable transformed plants. However, the amounts of the recombinant allergen were sufficient for analysis but not high enough to allow an economic production. Although molecular pharming has been shown to work well for the production of non-plant therapeutic proteins, it might be less efficient for closely related plant proteins.Electronic supplementary materialThe online version of this article (doi:10.1007/s00299-011-1199-3) contains supplementary material, which is available to authorised users.

Polymeric Structure and Host Toll-like Receptor 4 Dictate Immunogenicity of NY-ESO-1 Antigen in Vivo

In search of intrinsic factors that contribute to the distinctively strong immunogenicity of a non-mutated cancer/testis antigen, we found that NY-ESO-1 forms polymeric structures through disulfide bonds. NY-ESO-1 binding to immature dendritic cells was dependent on its polymeric structure and involved Toll-like receptor-4 (TLR4) on the surface of immature dendritic cells in mouse and human. Gene gun-delivered plasmid encoding the wild-type NY-ESO-1 readily induced T cell-dependent antibody (Ab) responses in wild-type C57BL/10 mice but not TLR4-knock-out C57BL/10ScNJ mice. Disrupting polymeric structures of NY-ESO-1 by cysteine-to-serine (Cys-to-Ser) substitutions lead to diminished immunogenicity and altered TLR4-dependence in the induced Ab response. To demonstrate its adjuvant effect, NY-ESO-1 was fused with a major mugwort pollen allergen Art v 1 and a tumor-associated antigen, carbonic anhydrase 9. Plasmid DNA vaccines encoding the fusion genes generated robust immune responses against otherwise non-immunogenic targets in mice. Polymeric structure and TLR4 may play important roles in rendering NY-ESO-1 immunogenic and thus serve as a potent molecular adjuvant. NY-ESO-1 thus represents the first example of a cancer/testis antigen that is a also damage-associated molecular pattern.

Sensitization Prevalence, Antibody Cross-Reactivity and Immunogenic Peptide Profile of Api g 2, the Non-Specific Lipid Transfer Protein 1 of Celery

BackgroundCelery (Apium graveolens) represents a relevant allergen source that can elicit severe reactions in the adult population. To investigate the sensitization prevalence and cross-reactivity of Api g 2 from celery stalks in a Mediterranean population and in a mouse model.Methodology786 non-randomized subjects from Italy were screened for IgE reactivity to rApi g 2, rArt v 3 (mugwort pollen LTP) and nPru p 3 (peach LTP) using an allergen microarray. Clinical data of 32 selected patients with reactivity to LTP under investigation were evaluated. Specific IgE titers and cross-inhibitions were performed in ELISA and allergen microarray. Balb/c mice were immunized with purified LTPs; IgG titers were determined in ELISA and mediator release was examined using RBL-2H3 cells. Simulated endolysosomal digestion was performed using microsomes obtained from human DCs.ResultsIgE testing showed a sensitization prevalence of 25.6% to Api g 2, 18.6% to Art v 3, and 28.6% to Pru p 3 and frequent co-sensitization and correlating IgE-reactivity was observed. 10/32 patients suffering from LTP-related allergy reported symptoms upon consumption of celery stalks which mainly presented as OAS. Considerable IgE cross-reactivity was observed between Api g 2, Art v 3, and Pru p 3 with varying inhibition degrees of individual patients' sera. Simulating LTP mono-sensitization in a mouse model showed development of more congruent antibody specificities between Api g 2 and Art v 3. Notably, biologically relevant murine IgE cross-reactivity was restricted to the latter and diverse from Pru p 3 epitopes. Endolysosomal processing of LTP showed generation of similar clusters, which presumably represent T-cell peptides.ConclusionsApi g 2 represents a relevant celery stalk allergen in the LTP-sensitized population. The molecule displays common B cell epitopes and endolysosomal peptides that encompass T cell epitopes with pollen and plant-food derived LTP.

Abstract 2696: Polymeric structure and host Toll-Like Receptor 4 dictate the immunogenicity of NY-ESO-1 in vivo

Abstract Introduction: NY-ESO-1 is a tumor-associated Ag (TAA) with distinctively strong immunogenicity. Our recent investigation of the specific interaction between NY-ESO-1 and complement C1q receptor (or Calreticulin, CRT) on the surface of immature DC, macrophages, and monocytes, indicated a unique interaction between NY-ESO-1 and the innate immune system (The Journal of Immunology, Zeng et al 2006). We further explore the molecular mechanism of the link between NY-ESO-1 and immature DC. Experimental Procedures: Cysteine-to-serine (C-to-S) substitutions were introduced into NY-ESO-1 proteins. Wild type NY-ESO-1 and mutant proteins were expressed in bacterial and mammalian cells and detected by western blot. The binding properties of wild type and mutant NY-ESO-1 with human and mouse immature DC were characterized. Bone marrow derived immature DC from the wild-type, TLR2-/-, and TLR4-/- mice were used for binding to NY-ESO-1. Mouse prostate cancer myc-cap cells with NY-ESO-1 and variants expression were co-cultured with human immature DC and mouse macrophage cell RAW264, and Th1/Th2 cytokines secretion were measured. Normal and the TLR4-/- mice were immunized with gene-gun delivery of plasmids encoding the wild type and variants of NY-ESO-1. Plasmids encoding NY-ESO-1 fused with a major mugwort pollen allergen Art v1 and a TAA carbonic anhydrase 9 were also delivered by gene gun to balb/c mice for immunization. Results: Wild Type NY-ESO-1 showed clear formation of dimers, trimers and even polymers due to inter-molecular disulfide bonds. Mutant NY-ESO-1 with all cysteines substituted by serines appeared only as monomer. C-to-S mutations significantly abolished the ability of NY-ESO-1 binding to human and mouse immature DC in vitro. The absence of TLR4 but not TLR2 partially impaired the binding of NY-ESO-1 to mouse bone marrow derived DC in vitro. Similarly, binding of recombinant NY-ESO-1 to human immature DC was blocked by anti-TLR4 monoclonal antibody. Mouse TLR4 co-precipitated with NY-ESO-1 by western blotting confirming the direct interaction between NY-ESO-1 and TLR4. Disrupting polymeric structure of NY-ESO-1 led to diminished immunogenicity and altered TLR4 dependence in the induced antibody responses. NY-ESO-1 expression in myc-cap cells impacted the immature DC inflammatory responses in co-culture system. Myc-cap cells bearing membrane-anchored NY-ESO-1 also affected tumor growth in vivo, correlated with in vitro studies. Gene gun-delivered plasmid DNA encoding the fusion genes generated robust immune responses against otherwise non-immunogenic targets in mice. Conclusion: NY-ESO-1 binds to immature DC depending on its polymeric architectural structure and its interaction with immature DC surface receptor TLR4. The unique interaction between NY-ESO-1 and immature DC renders NY-ESO-1 serve as a molecular adjuvant. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2696. doi:10.1158/1538-7445.AM2011-2696

Allergic rhinitis (AR) in geriatric patients

Allergic rhinitis (AR) can be defined as an inflammatory disease of the nose and the paranasal sinuses, characterized by a specific IgE-mediated hypersensitivity reaction. The aim of this study was to evaluate the correlation between the symptoms of AR and the prick test results in geriatric patients presenting with symptoms of AR by comparing these with those of a young control group. Thirty-two geriatric patients (Group 1) were analyzed retrospectively, and 37 patients (Group 2) were selected as the control group. Diagnosis of AR was made based upon the physical examination findings, nasal endoscopic examination findings and the skin prick test results. While the skin prick test positivity was 50% in Group 1, this rate was found as 75.7% in Group 2. The difference was found to be statistically significant (p=0.044). A statistically significant difference was found between the two groups in terms of susceptibility to mugwort pollen and fish (p=0.048, p=0.033). In conclusion, in geriatric patients presenting with AR symptoms, systemic treatment should not be initiated before performing skin prick test, due to the adverse effects of the drugs.

Molecular characterization of Api g 2, a novel allergenic member of the lipid‐transfer protein 1 family from celery stalks

Celery represents a relevant cross-reactive food allergen source in the adult population. As the currently known allergens are not typical elicitors of severe symptoms, we aimed to identify and characterize a non-specific lipid transfer protein (nsLTP). MS and cDNA cloning were applied to obtain the full-length sequence of a novel allergenic nsLTP from celery stalks. The purified natural molecule consisted of a single isoallergen designated as Api g 2.0101, which was recombinantly produced in Escherichia coli Rosetta-gami. The natural and recombinant molecules displayed equivalent physicochemical and immunological properties. Circular dichroism revealed a typical α-helical fold and high thermal stability. Moreover, Api g 2 was highly resistant to simulated gastrointestinal digestion. As assessed by ELISA, thermal denaturation did not affect the IgE binding of Api g 2. Natural and recombinant Api g 2 showed similar allergenic activity in mediator release assays. Api g 2-specific IgE antibodies cross-reacted with peach and mugwort pollen nsLTPs. Based on our results, it can be anticipated that inclusion of recombinant Api g 2 in the current panel of allergens for molecule-based diagnosis will facilitate the evaluation of the clinical relevance of nsLTP sensitization in celery allergy and help clinicians in the management of food allergic patients.

Mugwort-Sensitized Individuals from North Europe, South Europe and North America Show Different IgE Reactivity Patterns

Background: Sensitization to weed pollen allergens at the molecular level is not fully understood. We studied IgE reactivity to the major mugwort allergen Art v 1 in relation to allergens from other weed pollen and cross-reactive components in mugwort-sensitized subjects. Methods: Art v 1 and Amb a 1 were affinity purified and coupled to experimental ImmunoCAP® tests. Samples from North Europe (n = 50), South Europe (n = 19) and North America (n = 41) were analyzed for IgE against mugwort pollen, weed allergen components, pan-allergens and cross-reactive carbohydrate determinants (CCDs). Results: The prevalence of IgE reactivity (>0.35 kU_A/l) to Art v 1 was significantly higher in samples from North Europe than in those from North America. IgE to Amb a 1 was more common in North America than in North and South Europe, while IgE to Par j 2 was common in South Europe, less common in North America, and absent in North Europe. IgE to Art v 3 in mugwort-allergic patients was more common in North Europe than in South Europe and North America, while IgE to Sal k 1 was similar between the areas. Subjects with an Art v 1/mugwort-specific IgE ratio <0.5 had more often IgE to Amb a 1, profilin, polcalcin and CCDs than subjects with a ratio >0.5. Conclusions: Mugwort-sensitized subjects have different IgE reactivity profiles to weed allergens, reflecting their exposure to various pollens. Subjects with a low ratio between the IgE levels to Art v 1 and mugwort have a diverse IgE reactivity profile, indicating a role for cross-reactive allergens in their mugwort sensitization.

A New Allergen from Ragweed (Ambrosia artemisiifolia) with Homology to Art v 1 from Mugwort

Art v 1, the major pollen allergen of the composite plant mugwort (Artemisia vulgaris) has been identified recently as a thionin-like protein with a bulky arabinogalactan-protein moiety. A close relative of mugwort, ragweed (Ambrosia artemisiifolia) is an important allergen source in North America, and, since 1990, ragweed has become a growing health concern in Europe as well. Weed pollen-sensitized patients demonstrated IgE reactivity to a ragweed pollen protein of apparently 29-31 kDa. This reaction could be inhibited by the mugwort allergen Art v 1. The purified ragweed pollen protein consisted of a 57-amino acid-long defensin-like domain with high homology to Art v 1 and a C-terminal proline-rich domain. This part contained hydroxyproline-linked arabinogalactan chains with one galactose and 5 to 20 and more alpha-arabinofuranosyl residues with some beta-arabinoses in terminal positions as revealed by high field NMR. The ragweed protein contained only small amounts of the single hydroxyproline-linked beta-arabinosyl residues, which form an important IgE binding determinant in Art v 1. cDNA clones for this protein were obtained from ragweed flowers. Immunological characterization revealed that the recombinant ragweed protein reacted with >30% of the weed pollen allergic patients. Therefore, this protein from ragweed pollen constitutes a novel important ragweed allergen and has been designated Amb a 4.

Mapping the Interactions between a Major Pollen Allergen and Human IgE Antibodies

The interaction of specific IgE antibodies with allergens is a key event in the induction of allergic symptoms, thus representing an important target for therapeutic interventions in Type I allergies. We report here the solution NMR structure of Art v 1, the major mugwort pollen allergen. Art v 1 is the first protein structure with an allergenic defensin fold linked to a polyproline domain, which has not been identified in any reported allergen structure in the PDB. Moreover, the direct interaction of polyclonal IgE antibodies from an allergic patient has been mapped on the surface of an allergen for the first time. The data presented herein provide the basis for the design of tools for safe and effective vaccination against mugwort pollen allergy.

Rape and buckwheat honey allergy in relation to heat treatment

Introduction. Rape and buckwheat are progressively cultured in Lithuania. Pollen of these plants can be present in honeybee products, whereas it is known that pollen can be a potential allergen. Purpose. To explore how often and intensively the pollen of rape, buckwheat and mugwort, as well as monofloral (buckwheat and rape) and polyfloral (spring) honey can cause allergy in humans, to reveal whether there is a difference between honey allergy before and after heat treatment and to investigate the purity of monofloral honey in Lithuania. Materials and methods. The influence of allergens was determined by a skin-prick test. The botanical origin of honey and pollen was determined by the melissopalynological method. Results. Botanical composition studies have shown that in Lithuania where melliferous plants are cultivated, honey made by honeybees is almost homogeneous. The skinprick test revealed no statistically significant difference between honey tested before and after heat-treatment, but proved that monofloral honey was less allergenic than polyfloral. Pollen caused allergy more often and more intensively than monofloral honey did. Keywords: pollen, honey, allergy, heat treatment, skin-prick test

Correlation of oral allergy syndrome due to plant-derived foods with pollen sensitization in Japan

The clinical features of patients with oral allergy syndrome (OAS) due to plant-derived foods related to pollen allergy have been rarely reported in Japan. To evaluate the characteristics of OAS and to investigate whether pollen sensitizations are correlated with the prevalence of OAS in Japan. We measured specific IgE antibodies against 5 pollens in 622 outpatients (277 males and 345 females; mean age, 37 years) with atopic dermatitis, urticaria, angioedema, and food allergy during a 2-year period. Furthermore, OAS is diagnosed based on anamnesis and positive skin prick test reactions to suspected foods. Eighteen of the 436 patients (4.1%) sensitized to pollens were diagnosed as having OAS. Rates of specific IgE antibody-positive responses against Japanese cedar, ragweed, orchard grass, mugwort, and alder pollen were 69.8%, 35.3%, 29.1%, 24.1%, and 19.6%, respectively. The prevalence of OAS showed a significant positive correlation with sensitization to alder (P < .001). In addition, the most frequent causative foods were found to be apple, peach, and melon. The prevalence of OAS due to apple showed a significant positive correlation with sensitization to alder (P < .001) pollen, due to peach showed a significant positive correlation with sensitization to alder (P < .001) and orchard grass (P < .05) pollen, and due to melon showed a significant positive correlation with sensitization to alder (P < .005), orchard grass (P < .05), and ragweed (P < .05) pollen. Sensitization to pollens from species in the Betulaceae family is most strongly implicated in causing OAS in Japan.

口腔アレルギー症候群におけるシラカバ花粉リコンビナントアレルゲンに対するＩｇＥ抗体価

Persons allergic to birch pollen often report oral and pharyngeal hypersensitivity to fruit and vegetables, such as apples and peaches due to immunological cross-reactivity, or oral allergy syndrome (OAS) sometimes accompanied by systemic reaction. Such cross-reactive antigen reactions involve Bet v 1, the main birch-pollen allergen, and Bet v 2, birch-pollen profilin. We evaluated the food/antigen relationship. Subjects interviewed numbered 60-40 women and 20 men aged 12 to 70 (mean age: 35 years)-suffering OAS episodes and having IgE birch-pollen antibodies. Using CAP scoring we examined IgE antibodies to recombinant Bet v 1 (rBet v 1), recombinant Bet v 2 (rBet v 2), and recombinant Pru p 3 (rPru p 3) a peach lipid transfer protein (LTP). A CAP score of 0.35 or more was considered positive. We evaluated the relationship between recombinant allergens and 9 fruit often involving OAS--apple, peach, cherry, kiwi, pear, melon, plum, strawberry, and watermelon-based on subjects' reports. Of the 60, all (100%) were rBet v 1-positive, 9 (15%) rBet v 2-positive, and none (0%) rPru p 3-positive. Rose-family fruit-apples, peaches, cherries, pears, plums, and strawberries-often caused OAS regardless of positive or negative rBet v 2 CAP and were associated with rBet v 1. In contrast, more of those who were rBet v 2 CAP-positive had OAS to non-rose-family fruit-melon and watermelon-than those rBet v 2-negative. In rose-family and non-rose-family classification of the 9 fruit, cluster analysis and kappa statistics showed non-rose-family melon, watermelon, and kiwi to be associated with rBet v 2, as were grass and mugwort pollen allergies. Bet v 1 is associated with OAS due to rose-family fruit and Bet v 2 with OAS due to non-rose-family fruit.

Targeting the cysteine-stabilized fold of Art v 1 for immunotherapy of Artemisia pollen allergy

Plants of the genus Artemisia domestic in Northern and Central Europe, USA and parts of Asia are a major cause of allergic symptoms from late summer to autumn. Art v 1, the major mugwort pollen allergen appears as two-domain glycoprotein, consisting of an N-terminal defensin-like and a proline/hydroxyproline-rich C-terminal part. Patients sensitized to Art v 1 commonly display IgE antibodies against the cysteine-stabilized defensin fold. Site-directed mutagenesis of eight cysteines was used to disrupt disulfide bonds to generate molecules with altered IgE-binding capacity. Engineered constructs were expressed in E. coli and recombinant proteins were tested for their allergenic and T cell reactivity as well as for their physicochemical characteristics. Three cysteine variants (C22S, C47S, and C49S) exhibited extremely low IgE-binding activity in immunoblot and ELISA using sera from Art v 1-allergic patients. Mediator release assays using rat basophil leukemia cells showed that these variants displayed a 1×105-fold reduced allergenic potency as compared to wild-type protein. All variants were able to activate allergen-specific T cells in PBMC, as well as Art v 1-specific T cell lines and clones. Variant C49S displayed an increased hydrophobic surface potential which correlated with an advanced activation of allergen-specific T cells. The low allergenicity and high immunogenic activity of Art v 1 variant C49S renders the molecule an attractive candidate for hypoallergen-based immunotherapy of Artemisia pollen allergy.