Progress and gaps in understanding mechanisms of ash tree resistance to emerald ash borer, a model for wood-boring insects that kill angiosperms.

We review the literature on host resistance of ash to emerald ash borer (EAB, Agrilus planipennis), an invasive species that causes widespread mortality of ash. Manchurian ash (Fraxinus mandshurica), which coevolved with EAB, is more resistant than evolutionarily naïve North American and European congeners. Manchurian ash was less preferred for adult feeding and oviposition than susceptible hosts, more resistant to larval feeding, had higher constitutive concentrations of bark lignans, coumarins, proline, tyramine and defensive proteins, and was characterized by faster oxidation of phenolics. Consistent with EAB being a secondary colonizer of coevolved hosts, drought stress decreased the resistance of Manchurian ash, but had no effect on constitutive bark phenolics, suggesting that they do not contribute to increased susceptibility in response to drought stress. The induced resistance of North American species to EAB in response to the exogenous application of methyl jasmonate was associated with increased bark concentrations of verbascoside, lignin and/or trypsin inhibitors, which decreased larval survival and/or growth in bioassays. This finding suggests that these inherently susceptible species possess latent defenses that are not induced naturally by larval colonization, perhaps because they fail to recognize larval cues or respond quickly enough. Finally, we propose future research directions that would address some critical knowledge gaps.

Fraxinus pollen and allergen concentrations in Ourense (South-western Europe).

In temperate zones of North-Central Europe the sensitization to ash pollen is a recognized problem, also extended to the Northern areas of the Mediterranean basin. Some observations in Switzerland suggest that ash pollen season could be as important as birch pollen period. The allergenic significance of this pollen has been poorly studied in Southern Europe as the amounts of ash pollen are low. Due to the high degree of family relationship with the olive pollen major allergen (backed by a sequence identity of 88%), the Fraxinus pollen could be a significant cause of early respiratory allergy in sensitized people to olive pollen as consequence of cross-reactivity processes. Ash tree flowers in the Northwestern Spain during the winter months. The atmospheric presence of Ole e 1-like proteins (which could be related with the Fra a 1 presence) can be accurately detected using Ole e 1 antibodies. The correlation analysis showed high Spearman correlation coefficients between pollen content and rainfall (R(2)=-0.333, p<0.01) or allergen concentration and maximum temperature (R(2)=-0.271, p<0.01). In addiction CCA analysis showed not significant differences (p<0.05) between the component 1 and 2 variables. PCFA analysis plots showed that the allergen concentrations are related to the presence of the Fraxinus pollen in the air, facilitating the wind speed its submicronic allergen proteins dispersion. In order to forecast the Fraxinus allergy risk periods, two regression equations were developed with Adjusted R(2) values around 0.48-0.49. The t-test for dependent samples shows no significant differences between the observed data and the estimated by the equations. The combination of the airborne pollen content and the allergen quantification must be assessed in the epidemiologic study of allergic respiratory diseases.

Rising Out of the Ashes: Additive Genetic Variation for Crown and Collar Resistance to Hymenoscyphus fraxineus in Fraxinus excelsior.

Since the early 1990s, ash dieback due to the invasive ascomycete Hymenoscyphus fraxineus is threatening Fraxinus excelsior in most of its natural range. Previous studies reported significant levels of genetic variability in susceptibility in F. excelsior either in field or inoculation experiments. The present study was based on a field experiment planted in 1995, 15 years before onset of the disease. Crown and collar status were monitored on 777 trees from 23 open-pollinated progenies originating from three French provenances. Health status was modeled using a Bayesian approach where spatiotemporal effects were explicitly taken into account. Moderate narrow-sense heritability was found for crown dieback (h2 = 0.42). This study is first to show that resistance at the collar level is also heritable (h2 = 0.49 for collar lesions prevalence and h2 = 0.42 for their severity) and that there is significant genetic correlation (r = 0.40) between the severities of crown and collar symptoms. There was no evidence for differences between provenances. Family effects were detected, but computing individual breeding values showed that most of the genetic variation lies within families. In agreement with previous reports, early flushing correlates with healthier crown. Implications of these results in disease management and breeding are discussed.

Characterization of profilin and polcalcin panallergens from ash pollen.

BACKGROUND: Ash (Fraxinus excelsior) is an important source of allergenic pollen in temperate areas of Europe. Profilin and polcalcin are 2 important panallergens involved in cross-reactivity between different sources. OBJECTIVE: To clone and produce Fra e 2 (profilin) and Fra e 3 (polcalcin) as recombinant proteins and evaluate their immunological properties using the natural forms obtained from ash pollen. METHODS: Total RNA from ash pollen was used as a template to obtain the specific complementary DNA (cDNA) sequences of the 2 panallergens. The cDNA-encoding sequences were cloned into the pET11b expression vector and used to transform BL21 (DE3) Escherichia coli cells. Proteins were expressed, purified by chromatography, and characterized structurally by circular dichroism, mass spectrometry, and immunologically by western blot and ELISA using profilin and polcalcin polyclonal antibodies and human sera from ash pollen-sensitized patients. RESULTS: Profilin and polcalcin amino acid sequences from ash pollen showed a high degree of identity with homologous allergens from different sources. The cDNA-encoding allergen sequences were expressed as nonfusion recombinant proteins and purified to homogeneity. Secondary structure values were similar to those obtained from other members of these families. Allergenic properties of the recombinant allergens were observed to be equivalent to those of the natural counterparts of F excelsior pollen. CONCLUSIONS: Fra e 2 and Fra e 3 recombinant allergens might be used in clinical diagnosis to determine profilin- and polcalcin-specific IgE levels present in the sera of ash pollen-sensitized patients, thus facilitating the finding of the sensitizing source in areas with complex sensitization profiles.

Presence of natural genetic resistance in Fraxinus excelsior (Oleraceae) to Chalara fraxinea (Ascomycota): an emerging infectious disease.

Fraxinus excelsior, common ash native to Europe, is threatened by a recently identified pathogenic fungus Chalara fraxinea, which causes extensive damage on ash trees across Europe. In Denmark, most stands are severely affected leaving many trees with dead crowns. However, single trees show notably fewer symptoms. In this study, the impact of the emerging infectious disease on native Danish ash trees is assessed by estimating presence of inherent resistance in natural populations. Disease symptoms were assessed from 2007 to 2009 at two different sites with grafted ramets of 39 selected clones representing native F. excelsior trees. A strong genetic variation in susceptibility to C. fraxinea infections was observed. No genetic or geographic structure can explain the differences, but strong genetic correlations to leaf senescence were observed. The results suggest that a small fraction of trees in the Danish population of ash possess substantial resistance against the damage. Though this fraction is probably too low to avoid population collapse in most natural or managed ash forests, the observed presence of putative resistance against the emerging infectious disease in natural stands is likely to be of evolutionary importance. This provides prospects of future maintenance of the species through natural or artificial selection in favour of remaining healthy individuals.

Oleaceae-induced pollinosis in an area with exposure to olive and ash trees.

BACKGROUND: Navarre, in Northern Spain, is an area with moderate exposure to olive and ash tree pollen. OBJECTIVE: To assess the relevance of ash as a cause of pollinosis in our region. METHODS: The study sample comprised 85 patients from Navarre with clinical symptoms of pollinosis. Specific immunoglobulin E (sIgE) was determined to Fra e 1, Ole e 1, and a mixture of amino- and carboxy-terminal domains of Ole e 9 (Ole e 9 NC) (ADVIA-Centaur). At the same time, the presence of sIgE to other pollen allergens was studied. Prick tests were performed with ash pollen (n=33) and olive pollen (n=85) and the symptomatic period was recorded (n=85). As a control group, we studied the serum of 98 patients with olive pollen allergy, intense exposure to olive pollen, and no exposure to ash. RESULTS: Sensitization to Oleaceae was detected in 24/85 patients in the study group (28.2%). In this group, the mean (SD) level of IgE to Fra e 1 was 8.5 (10) kU(A)/L and to Ole e 16.07 (7.88) kU(A)/L (P < .001). In the control group, these figures were 103.64 (132.19) kU(A)/L and 86.43 (118.5) kU(A)/L (P < .001), respectively. In all patients with positive sIgE to Fra e 1, IgE to Ole e 1 was also detected (concordance index, kappa = 1), both in the study group and in the control group. Patients who were sensitized to Fra e 1 did not present a longer symptomatic period and their symptoms did not have an earlier onset. CONCLUSION: We did not find evidence of clinically relevant sensitization to ash in Navarre.

Evaluation of ash pollen sensitization pattern using proteomic approach with individual sera from allergic patients.

BACKGROUND: In Europe, sensitization to ash pollen induces pollinosis with cross-reactivities with other pollen sources. The aim of the study was to identify the repertoire of ash pollen allergens and evaluate the extent of the diversity of the IgE response in ash allergic patients. METHODS: The IgE reactivities of 114 ash pollen- and eight grass pollen-sensitized patients were screened by 1D immunoblot (SDS-PAGE) against ash pollen extract. The IgE reactivities of 13 ash pollen- and two grass pollen-sensitized patients were then evaluated in 2D immunoblots. Some IgE- and non-IgE-reactive proteins were identified by mass spectrometry. RESULTS: In 1D analysis, 86% of sera showed binding to Fra e 1 (18-20 kDa), 23% to Fra e 2 (14 kDa), 3% to Fra e 3 (10 kDa) and 57% to High Molecular Weight allergens (HMW, >30 kDa). Individual analysis of 2D immunoblots showed several IgE-binding protein areas among which three were more often recognized: (i) Fra e 1 comprising, at least, 15 isoforms, (ii) a series of acidic spots (45 kDa), and (iii) Fra e 2, the ash profilin. HMW allergens could be resolved in four areas; two unidentified, one homologous to beta-galactosidase and the other to sugar transport proteins. A malate deshydrogenase and calmodulin were shown to be IgE-binding proteins and 10 non-IgE reactive proteins were identified. CONCLUSIONS: No direct correlation was evidenced between IgE profile and the degree of sensitization even though 2 spectrotypes could be distinguished. Our data contribute to a better delineation of ash pollen allergens and patterns of sensitization.

Short-term effects of airborne pollens on asthma attacks as seen by general practitioners in the Greater Paris area, 2003-2007.

AIMS: To investigate for the first time the short-term effects of airborne pollen counts on general practitioner (GP) consultations for asthma attacks in the Greater Paris area between 2003-2007. METHODS: Counts were available for common pollens (Betula, Cupressa, Fraxinus and Poaceae). Weekly data on GP visits for asthma attacks were obtained from the French GP Sentinel Network. A quasi-Poisson regression with generalised additive models was implemented. Short-term effects of pollen counts were assessed using single and multi-pollen models after adjustment for air pollution and influenza. RESULTS: A mean weekly incidence rate of 25.4 cases of asthma attacks per 100,000 inhabitants was estimated during the study period. The strongest significant association between asthma attacks and pollen counts was registered for grass (Poaceae) in the same week of asthma attacks, with a slight reduction of the effect observed in the multi-pollen model. Adjusted relative risk for Poaceae was 1.54 (95% CI: 1.33-1.79) with an inter-quartile range increase of 17.6 grains/m3 during the pollen season. CONCLUSIONS: For the first time, a significant short-term association was observed between Poaceae pollen counts and consultations for asthma attacks as seen by GPs. These findings need to be confirmed by more consistent time-series and investigations on a daily basis.

Airborne pollen of Carya, Celtis, Cupressus, Fraxinus and Pinus in the metropolitan area of Monterrey Nuevo Leon, Mexico.

The concentration of pollen grains in the atmosphere over the metropolitan area of Monterrey, Nuevo Leon, Mexico, was analyzed throughout a year from March 2003-February 2004, focused on the genus Carya, Celtis, Cupressus, Fraxinus and Pinus owing to their interest as etiological pollinosis agents in diverse regions of the world. A 7-day Hirst type volumetric spore and pollen trap was located on a building roof of the city at 15 m from ground level for continuous sampling. The total quantity of pollen recorded for the study period was 21,083 grains/m(3), corresponding to 49.75 % of the taxa of interest. February and March were the months with higher pollen amounts in the air with 7,525 and 2,781 grains/m(3), respectively, and amounted to 49 % of total year through pollen. Fraxinus was the genus which contributed to the largest amount of pollen with 28 % of total grains (5,935 grains/m(3)) followed by Cupressus with 13 % (2,742 grains/ m(3)). Celtis, Pinus and Carya contributed with 5.3 % , 2.7 % , and 0.6 % of total pollen, respectively. These results indicate that Fraxinus and Cupressus are present in the area in sufficient quantity to indicate likely involvement in the origin of allergic disorders in the human population.

Development of new sensitizations in asthmatic children monosensitized to house dust mite by specific immunotherapy.

It has been hypothesized that specific immunotherapy (SIT) significantly decreases the development of new allergen sensitizations in mono-sensitized patients. In this study, we evaluated the effect of SIT on the development of new allergen sensitizations in 129 asthmatic children mono-sensitized to house dust mite. SIT was accepted by only 70 of them (SIT group). The remaining 59 children were treated only with medication (control group). At the end of the study we found that 33% of all patients developed new sensitizations. Surprisingly, the prevalence of new sensitizations was significantly higher in the SIT group (45.5%) than in the control group (18.1 %). Ash tree (Fraxinus excelsior), Olive and Meadow fescue (Festuca elatior) were the most common allergens responsible for the new sensitizations. We conclude that SIT did not prevent the onset of new sensitizations in asthmatic children mono-sensitized to house dust mite.

Potential health risk of allergenic pollen with climate change associated spreading capacity: Ragweed and olive sensitization in two German federal states.

BACKGROUND: Global climate changes may influence the geographical spread of allergenic plants thus causing new allergen challenges. OBJECTIVE: Allergy patients from two German federal states were compared for their status quo sensitization to ragweed, an establishing allergen, olive, a non-established allergen, and the native allergens birch, mugwort, and ash. METHODS: Between 2011 and 2013, 476 adult allergy patients per region were recruited. Patients completed a questionnaire, participated in a medical interview, and underwent skin prick testing and blood withdrawal for analysis of specific IgE to allergen components (ISAC technology). Data on regional pollen load from 2006 to 2011 were acquired from the German Pollen Information Service Foundation. RESULTS: Prick test reactivity to ragweed and ash, respectively, was lower in Bavaria than in NRW (ragweed: p=0.001, aOR=0.54; ash: p=0.001, aOR=0.59), whereas prick test reactivity to olive was higher (p=0.000, aOR=3.09). Prick test reactivity to birch and mugwort, respectively, did not significantly differ. 1% (1/127) of patients with prick test reactivity to ragweed showed sIgE to Amb a 1, and 65% (86/132) of olive-but-not-ash reactive patients showed sIgE to Ole e 1 (NRW: 67%, Bavaria: 65%; p=0.823, OR=0.91). Regional differences in sensitization pattern were neither explainable by cross-reactivity to pollen pan-allergens nor non-exposure variables nor by reported plant population or pollen data. CONCLUSIONS: Spread of ragweed and particularly olive may result in prompt occurrence of allergic symptoms. Early identification of invasive allergens due to climate change does need time and spatial close meshed measurement of respective indicator allergens and sensitization pattern.

Prevalence of ragweed allergy in rural Geneva - a pilot study.

OBJECTIVE: The prevalence of ragweed allergy is increasing worldwide. Ragweed distribution and abundance is spreading in Europe in a wide area ranging from the Rhone valley in France to Hungary and Ukraine, where the rate of the prevalence can peak at as high as 12%. Low-grade ragweed colonisation was seen in Geneva and Ticino, less than two decades ago. There were fears that allergies to ragweed would increase Switzerland. The intent of this study was to assess the rate of prevalence of sensitisation and allergy to ragweed in the population living in the first rural Swiss setting where ragweed had been identified in 1996, and to evaluate indirectly the efficacy of elimination and containment strategies. MATERIAL AND METHODS: In 2009, 35 adults in a rural village in the Canton of Geneva were recruited. Data were collected by means of questionnaires and skin-prick tests were done on each participant. The study was approved by the local Ethics Committee. RESULTS: Based on questionnaires, 48.6% had rhinitis (95% confidence interval [CI] 32.9-64.4; n = 17/35) and 17.1% asthma (95% CI 8.1-32.6; n = 6/35). Atopy was diagnosed in 26.4% (95% CI 12.9-44.4) of the sample (n = 9/34). Ragweed sensitisation was found in 2.9% (95% CI 0.7-19.7; n = 1/34), mugwort sensitisation in 2.9% (95% CI 0.1-14.9; n = 1/35), alder sensitisation in 17.1% (95% CI 6.6-33.6; n = 6/35), ash sensitisation in 12.5% (95% CI 3.5-29.0; n = 4/32) and grass sensitisation in 22.9% (95% CI 10.4-40.1; n = 8/35). Ragweed (95% CI 0.1-14.9; n = 1/34) and mugwort allergies (95% CI 0.1-14.9; n = 1/35) were both found in 2.9% of the population. CONCLUSION: This study showed a surprisingly low incidence of ragweed sensitisation and allergy, of 2.9% and 2.9%, respectively, 20 years after the first ragweed detection in Geneva. The feared rise in ragweed allergy seems not to have happened in Switzerland, compared with other ragweed colonised countries. These results strongly support early field strategies against ragweed.

[Polyclonal antisera to birch pollen allergen for the air allergen screening]. / Poliklonal'nye antisyvorotki protiv allergena pyl'tsy berezy dlia skrininga aéroallergenov.

The cross reactivity of the pollen allergens of birch (Betula pendula), hazel (Corylus avelana), alder (Alnus glutinosa), wormwood (Artemisia absinthium), oak (Quercus robur) and ashtree (Fraxinus excelsior) was studied by the methods of the enzyme immunoassay and gel diffusion with the use of rabbit polyclonal antisera (PAS) prepared against birch pollen allergen. PAS was shown to be greatly related to all above-mentioned allergens with the exception of oak pollen allergen. PAS was found capable of forming precipitates in the layer of agar gel only with birch pollen allergen. A proposal was made to use PAS, in view of the established specific features of its immunochemical properties, in the development of test systems for the ecological monitoring of biotopes.

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.

Changing the rules of the game.

Genomics researchers have built a Facebook game that allows members of the public to join the effort to understand a disease that has killed millions of ash trees across Europe.

Standardization of an ash (Fraxinus excelsior) pollen allergen extract.

BACKGROUND: Ash tree (Fraxinus excelsior) is the main representative of the Oleaceae family in temperate zones. Diagnosis of ash pollen allergy is made difficult due to (1) an overlapping pollinization period with Betulaceae, (2) non-inclusion in current diagnostic assays, and (3) some cross- reactivity with minor allergens from Betulaceae. The aim of this study was to calibrate an ash pollen in-house reference preparation (IHRP) in allergic patients in order to produce standardized products for diagnosis and immunotherapy purposes. METHODS: Ash pollen IHRP was extracted, ultrafiltered and freeze dried. Allergens in the extract were detected after 2-dimensional PAGE using specific sera and a monoclonal antibody. The Fra e 1 content of IHRP was evaluated by quantitative immunoprint. Forty-eight subjects from the North-East of France exhibiting clinical symptoms, a positive skin test and specific IgE levels > or =class 2 to ash pollen were recruited. IgE immunoprints were performed to select patients sensitized to the ash Fra e 1 allergen as opposed to cross-reacting allergens. Serial 10-fold dilutions of the IHRP were tested by skin prick tests in order to determine the concentration inducing a geometrical mean wheal diameter of 7 mm, said to correspond to an index of reactivity (IR) of 100 per millilitre. RESULTS: IgE-reactive molecules in IHRP comprise Fra e 1, Fra e 2, a 9-kDa molecule (presumably Fra e 3), as well as a doublet at 15 kDa and high molecular weight allergens. The 100 IR concentration of IHRP inducing a geometrical mean wheal diameter of 7 mm in 22 patients sensitized to Fra e 1 corresponds to the 1/126 (w/v) extraction ratio (i.e. 259 microg/ml of protein by Bradford) and contains 17 microg/ml of Fra e 1. The variability in total activity of 5 batches of standardized extracts was found to be significantly reduced when compared with 7 non-standardized extracts. CONCLUSION: An ash pollen IHRP was defined and molecularly characterized. Its successful standardization at 100 IR/ml in patients specifically sensitized to Fra e 1 allowed a skin reactivity-based calibration in properly diagnosed patients. Such a standardized ash pollen extract is a reliable tool to support immunotherapy of ash pollen allergy.

Short-term effect of pollen exposure on antiallergic drug consumption.

BACKGROUND: Several studies have investigated the association between pollen exposure and asthma emergency admissions, but only 2 have investigated the effect of airborne allergens on consultations for rhinitis or conjunctivitis and none has used drug consumption as the health indicator. OBJECTIVE: To analyze the short-term association between pollen exposure and antiallergic drug consumption in the urban area of Clermont-Ferrand, France, taking into account the potentially confounding effect of air pollution and meteorological factors. METHODS: We used the French health insurance database to select all individuals from the Clermont-Ferrand urban area having benefited from reimbursement for antiallergic treatment from January 1, 2000, through December 31, 2001, and from January 1, 2003, through December 31, 2004. An episode of treated allergic rhinitis, rhinosinusitus, or conjunctivitis (ARC) was defined as the association of an oral antihistamine and a local antiallergic drug on the same prescription. The relations between daily changes in pollen concentrations and daily changes in the number of treated ARC cases were analyzed using a Poisson regression model with penalized spline functions. RESULTS: The risk of treated ARC associated with an interquartile increase in pollen concentration increased significantly for Poaceae (5%, P < .001), Fraxinus (7%, P < .001), Betula (7%, P < .001), and Corylus (2%, P < .02). This increase was significant in all age groups for Poaceae and Fraxinus pollen and in people younger than 65 years for Betula pollen. The effect was mainly concentrated on the present day, except for Poaceae pollens, for which the risk remained significantly (P < .001) increased until 3 days lag time. CONCLUSIONS: This study showed a significant increase in treated ARC cases related to Poaceae, Fraxinus, and Betula. Specific risks are difficult to evaluate for species that share the same pollination period. Time-series studies based on drug consumption are useful to highlight and to supervise pollen-related diseases requiring ambulatory care.

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.