Diagnosis of <Emphasis Type="Italic">Chenopodium album</Emphasis> allergy with a cocktail of recombinant allergens as a tool for component-resolved diagnosis

Chenopodium album pollen is one of the main sources of pollen allergy in desert and semi-desert areas and contains three identified allergens, so the aim of this study is comparison of the diagnostic potential of C. album recombinant allergens in an allergenic cocktail and C. album pollen extract. Diagnostic potential of the allergenic cocktail was investigated in 32 individuals using skin prick test and obtained results were compared with the acquired results from C. album pollen extract. Specific IgE reactivity against the pollen extract and allergenic cocktail was determined by ELISA and western blotting tests. Inhibition assays were performed for the allergenic cocktail characterization. The exact sensitization profile of all patients was identified which showed that 72, 81 and 46% of allergic patients had IgE reactivity to rChe a 1, rChe a 2 and rChe a 3, respectively. Almost all of C. album allergic patients (30/32) had specific IgE against the allergenic cocktail. In addition, there was a high correlation between IgE levels against the allergenic cocktail and IgE levels against the pollen extract. The allergenic cocktail was able to completely inhibit IgE binding to natural Che a 1, Che a 2 and Che a 3 in C. album extract. In addition, positive skin test reactions were seen in allergic patients that tested by the allergenic cocktail. The reliable results obtained from this study confirmed that the allergenic cocktail with high diagnostic potential could be replaced with natural C. album allergen extracts in skin prick test and serologic tests.     

Evaluation of the age of onset of respiratory allergic symptomatology

Summary In order to evaluate the age of ouset of respiratory allergic symptoms we examined 2315 patients (656 children and 1659 adults) coming to our clinic as out patients. Each patient underwent the following diagnostic tests: anamnestic and clinical examination, skin-tests for the most common allergens and, when requested, Radioallergosorbent test (RAST) and specific and non specific BPT.We selected 278 children and 673 adults for whom it was possible to establish (on the basis of precise anamnestic considerations) the age at which the respiratory symptoms first appeared.Among these, 300 reacted only toParietaria pollen, 58 only to grass pollen, 420 only to house-dust mite, and 173 were not allergic.Patients reacting to multiple allergens were excluded from the analysis.The results of our study show a significant precocity of the onset of symptoms related to allergic sensitization to mites with peak level at 5 years.On the other hand, the onset of pollen-related symptoms was later on (peak level at 20–25 years forParietaria pollen and 30–35 years for grass pollen). For non allergic patients we did not find a definite age of onset.     

The suggestive association of eotaxin-2 and eotaxin-3 gene polymorphisms in Korean population with allergic rhinitis

The eotaxin gene family (eotaxin, eotaxin-2 and eotaxin-3) has been implicated in the recruitment of eosinophils, basophiles and Th2 lymphocytes that are central aspects of allergic diseases. To determine whether single-nucleotide polymorphisms (SNPs) of the eotaxin-2 and eotaxin-3 genes are associated with susceptibility to allergic rhinitis, we scanned 178 allergic rhinitis patients and 281 controls without allergic rhinitis using the direct sequencing and single-base extension (SBE) methods. We also calculated the haplotype frequencies between +179T>C and +275C>T of eotaxin-2 and +2497T>G of eotaxin-3 in both controls and allergic rhinitis patients. The haplotype frequency between controls and allergic rhinitis patients was suggestively associated (P=0.0001). The genotype frequencies of eotaxin-3 +2497T>G in allergic rhinitis patients were suggestively different from those in non-allergic rhinitis controls (P=<0.0007). Our results strongly suggest that the SNP of eotaxin-3 might be associated with susceptibility to allergic rhinitis.     

Glycoproteins are species-specific markers and major IgE reactants in grass pollens

Grass pollen allergic patients are concomitantly exposed and sensitized to pollens from multiple Pooideae (i.e. common grass) species. As such, they are currently desensitized by allergen‐specific immunotherapy using extracts made from mixes of pollens from Anthoxanthum odoratum, Dactylis glomerata, Lolium perenne, Phleum pratense and Poa pratensis. Herein, we demonstrate that species‐specific glycoprotein patterns are documented by 1D and 2D electrophoresis and Western blotting analysis, which can be used as an identity test for such pollens. Most allergens are glycoproteins bearing complex N‐glycans encompassing β1,2 xylose and α1,3 fucose glycoepitopes. Glycoepitope destruction using periodate oxidation has no impact on seric IgE reactivity in 75% atopic patients (n = 24). The latter have thus no significant IgE responses to carbohydrate‐containing epitopes. In contrast, periodate treatment strongly impairs IgE recognition of glycoallergens in 25% of patients tested, demonstrating the presence of carbohydrate‐specific IgE in those patients. While the clinical impact of carbohydrate‐specific IgE is still a matter of controversy, the presence of these IgE in the serum of many allergic patients illustrates the need for cross‐reacting carbohydrate epitope‐free recombinant allergens to develop relevant diagnostic tests. These data also support the pertinence of mixing multiple grass pollens to desensitize atopic patients, with the aim to broaden the repertoire of glycoepitopes in the vaccine, thus mimicking natural exposure conditions.     

Comparative aerobiology,allergenicity and biochemistry of three palm pollen grains in Calcutta,India

A Burkard volumetric trap was used at Salt Lake City, Calcutta, to record the occurrence and frequency of three common palm pollen, namely,Areca catechu, Borassus flabellifer andPhoenix sylvestris for two consecutive years (July 1988–June 1990). The meteorological factors responsible for the frequency of relevant airborne pollen grains were analysed. The allergenic potential of these pollen types was investigated by skin-prick tests on adult respiratory allergic patients. These were also chemically analysed in terms of total carbohydrate, lipid and soluble protein. Total soluble protein of the above types was used in 11% SDS-polyacrylamide gel electrophoresis to study the range of molecular components.     

The relationships between the concentrations of airborne pollen and allergic symptoms in Trieste (Northern Italy) in 1989

Summary Concentrations of airborne pollens recorded in Trieste in 1989 are evaluated in relation to allergic complaints in 113 patients with skin prick tests positive to one pollen species. Analysis of the result enable the two most important allergens to be confirmed as: Poaceae in 70.7% of patients and Parietaria in 18.6%. Few people had monosensitation to Compositae (Artemisia), Corylaceae and Fagaceae.Sympotoms are related to the flowering period when pollen levels climbed to daily averages of 15–20 grains/m³ but they started later than airborne peak concentrations and finished later than pollen decline. Pollen concentration recording can be a useful way to predict the clinical manifestation in sensitive patients but other factors are involved in determining symptoms like subjective mucosal reactions, polysensitization, patients living and working environment.     

Prevalence of allergenic pollen grains in the aerosol of the city of Calcutta,India: a two year study

The role of pollen grains as a causative agent of respiratory allergic disorders such as asthma and allergic rhinitis is common and very well established. The aim of this study was to assess the frequencies of airborne pollen in the Calcutta metropolis and to identify the taxa which cause significant amounts of sensitization. An aeropalynological survey of the atmosphere of Calcutta was carried out from 2004 to 2006. Skin tests were performed with a panel of the most common pollen types on local patients with clinical features of pollinosis. The meteorological factors responsible for the frequency of the pollen types were analysed. The results of monthly visits to the clinic by these patients were correlated with the monthly pollen counts of three dominant and perennial pollen taxa. The dominant pollen types were Trema (19%), Poaceae (12.98%), Casuarina (5.76%), Cocos (5.7%), Azadirachta (4.65%), Peltophorum (3.71%), Cyperaceae (3.68%), Delonix (3.18%) and Areca (2.56%). Total pollen concentration seems to have a significant positive correlation with temperature and wind speed whereas there was a negative correlation with humidity. Skin tests were most frequently found to be positive with the pollen of Poaceae (49%), Azadirachta (46%), Cocos (47%), Cyperaceae (35%), Peltophorum (33%), Areca (29%), Phoenix (26%), and Borassus (23%). A positive correlation occurred between visits to the clinic and monthly pollen count of Areca, Cocos, and Poaceae. This is the first study to design a pollen calendar for Calcutta city; it will provide useful data for enabling allergologists to achieve accurate diagnosis for patients with pollen hypersensitivity.     

Identification of methionine synthase (Sal k 3), as a novel allergen of <Emphasis Type="Italic">Salsola kali</Emphasis> pollen

Salsola kali pollen is a common cause of pollinosis during summer and early fall in desert and semi-desert regions. The aim of this study was the identification and characterization of Sal k 3, a new allergen from S. kali pollen. S. kali pollen extract was fractionated by SDS-PAGE and the allergenic profile was determined by IgE-immunoblotting using twelve S. kali allergic patients. Protein identification was carried out by the means of mass spectrometry. Using degenerated primers, two DNA fragments encoding N- and C-terminal domain of Sal k 3 were amplified by PCR, then cloned into the PTZ57R/T vector and sequenced. The open reading frame of Sal k 3 fragments were subcloned in the pET-32b(+) vector, expressed in E. coli, and purified by Ni²⁺ affinity chromatography. The IgE-binding capacity of rSal k 3 fragments was then studied by IgE-immunoblotting, inhibition assays, and skin prick tests. A 45-kDa allergen was identified as a fragment of the cobalamin-independent methionine synthase (MetE) by mass spectrometry and was detected in the sera of 8/12 (66.6%) of S. kali allergic patients. Moreover, inhibition assays demonstrated that the purified rSal k 3 fragments were similar to their counterparts in the crude extract. Sal k 3 represents a new allergen of S. kali pollen and seems to be an important allergenic compound in S. kali pollen.     

Aerobiological, clinical and immunobiochemical studies on Lantana camara pollen and cross-reactivity with other Verbenaceae pollen species

Airborne pollen is an important and potent source of aeroallergens. The aim of the study was to conduct a 2-year aerobiological survey in Calcutta, India, for knowing the concentration and seasonal periodicity of Lantana camara (LC) pollen. The sensitization due to this pollen among seasonal respiratory allergic patients and its chemical composition was studied. An aerobiological survey was conducted with a volumetric Burkard sampler from 2004 to 2006. Protein components of LC pollen were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis and IgE immunoblotting. Allergenic activities were determined by in vivo (skin prick test) and in vitro (enzyme-linked immunosorbent assay. In vitro inhibition tests were performed to evaluate cross-reactivity. LC pollen was present from March to May and from September to December contributing up to 10.5% to the total aeropollen load during peak month. Horizontal profile showed highest concentration for nearest (0.5 m) rotorod and it was decreased by half in a distance within 4.5–6.5 m from plot edge. LC pollen contained 7.5% carbohydrate, 19.3% lipid with proline and valine as dominant amino acid. Among 1,500 adult respiratory allergic patients tested, 7.93% showed higher level of positive reaction. IgE binding proteins of 22, 42, 45 and 95 kD were revealed. LC pollen showed remarkable cross-reactivity with other local Verbenaceae pollen taxa (Clerodendron viscosum, Tectona grandis and Vitex negundo). This is the first study on LC pollen regarding its aerobiological, clinical and immuno-biochemical aspects; it should be helpful for the diagnosis and therapy of patients susceptible to LC pollen.     

Proteomic analysis of <Emphasis Type="Italic">Parietaria judaica</Emphasis> pollen and allergen profiling by an immunoproteomic approach

Parietaria judaica pollen is a common cause of airway allergic disease in the Mediterranean area. Proteome analysis of mature Parietaria judaica pollen by two-dimensional gel electrophoresis (2-DE) and mass spectrometry has established the first reference proteome map of this weed. Proteins involved in a variety of cellular functions as well as the occurrence of allergens were detected. By using 2-DE and immunoblotting with sera from Parietaria judaica allergic patients we obtained a more detailed characterization of Parietaria judaica allergen profile so to improve our comprehension of the pathogenesis of pollen-induced allergic reaction.     

Occurrence of fungal spores in the respiratory tract and homes of patients with positive skin test to fungi

The occurrence of fungal spores in the upper respiratory tract and in the homes of 86 patients with allergic manifestations and positive skin tests for fungi has been investigated.Alternaria tenuis andCladosporium herbarum have been more frequently found to give positive reactions, followed byAspergillus mix,Candida albicans andPenicillium notatum. Cladosporium andAlternaria represented also the most common moulds isolated from nasal specimens. The prevalent fungi isolated from patients' environments wereCladosporium, Penicillium andAlternaria, occurring respectively in 93%, 78% and 72% of the homes sampled. An examination of individual cases have revealed a surprisingly close correlation, in 47 out of 86 patients, between positive skin test response and presence in high amount of the same fungi in patient's respiratory tract and home. In the majority of cases the fungi involved wereAlternaria and/orCladosporium.     

Occurrence of fungal spores in the respiratory tract and homes of patients with positive skin test to fungi

The occurrence of fungal spores in the upper respiratory tract and in the homes of 86 patients with allergic manifestations and positive skin tests for fungi has been investigated.Alternaria tenuis andCladosporium herbarum have been more frequently found to give positive reactions, followed byAspergillus mix,Candida albicans andPenicillium notatum. Cladosporium andAlternaria represented also the most common moulds isolated from nasal specimens. The prevalent fungi isolated from patients' environments wereCladosporium, Penicillium andAlternaria, occurring respectively in 93%, 78% and 72% of the homes sampled. An examination of individual cases have revealed a surprisingly close correlation, in 47 out of 86 patients, between positive skin test response and presence in high amount of the same fungi in patient's respiratory tract and home. In the majority of cases the fungi involved wereAlternaria and/orCladosporium.     

Lymphocyte responses to food antigens in food sensitive patients with allergic tension-fatigue syndrome

Scores of radio allergosorbent test (BAST) for cow's milk or buckwheat flour and proliferative responses of peripheral blood mononuclear cells (PBMCs) to bovine serum albumin and-lactoglobulin or buckwheat flour were measured in cow's milk or buckwheat flour sensitive patients with allergic tension-fatigue syndrome. In all 3 cow's milk sensitive patients with allergic tension-fatigue syndrome, RAST scores for cow's milk were negative or slightly positive, but PBMCs well responded to bovine serum albumin and-lactoglobulin, but not to ovalbumin. In a buckwheat flour sensitive patient with allergic tension-fatigue syndrome, RAST scores for buckwheat flour were negative, but PBMCs well responded to buckwheat flour, but not to ovalbumin, bovine serum albumin and-lactoglobulin. Conversely, in cow's milk or buckwheat flour sensitive patients with immediate allergic symptoms, RAST scores for offending foods were positive although PBMCs did not respond to offending food antigens. These results suggest that proliferative responses of PBMCs to food antigens are very useful for detection of offending foods in allergic tension-fatigue syndrome.     

Grifola frondosa extract and ergosterol reduce allergic reactions in an allergy mouse model by suppressing the degranulation of mast cells

ABSTRACT

The increasing number of patients suffering from allergic diseases is a global health problem. Grifola frondosa is an edible mushroom consumed as a health food in Asia, and has recently been reported to have anti-allergic effects. We previously reported that G. frondosa extract (GFE) and its active components, ergosterol and its derivatives, inhibited the antigen-induced activation of RBL-2H3 cells. Here, we demonstrated that GFE and ergosterol also had an inhibitory effect on the degranulation of bone marrow–derived mast cells (BMMCs) and alleviated anaphylactic cutaneous responses in mice. Using an air pouch-type allergic inflammation mouse model, we confirmed that oral administration of GFE and ergosterol suppressed the degranulation of mast cells in vivo. Our findings suggest that G. frondosa, including ergosterol as its active component, reduces type I allergic reactions by suppressing mast cell degranulation in mice, and might be a novel functional food that prevents allergic diseases.     

Pollinosis related to Zygophyllum fabago in a Mediterranean area

Zygophyllum fabago is a herbaceous plant withairborne pollen found widely in the MurciaRegion, in the Southeast of Spain. Although itsallergenicity has been recently reported,little is known of its involvement inpollinosis. Aerobiological study andsensitization in pollinotics weremeasured using a Hirst volumetric trap. Wehave measured the atmospheric concentrationsof this pollen and other allergenicpollen types in our region, between March 1993 andMarch 1997. Z. fabago pollen wascollected for a morphometric study of thepollen grain, and a lyophilized extract wasprepared for skin prick tests. We haveconducted skin tests with different pollen typesfrom our region and with Z. fabago in1736 patients with symptomssuggesting pollinosis. The size of the pollengrain averages 15.17 × 17.35 µm. Thepollination period extends from May to August,with a mean accumulated concentration of 448grains/m³. Out of 1736 pollinotics,263 (15.15%) showed a positive skin test forZ. fabago, 6 were monosensitized and 257were sensitized to other common pollen types fromour Region. Specific IgE to Z. fabago wasequal or higher than 0.35 ku/l in 86.56% ofsensitized patients. Chenopodiaceae pollinateduring spring and autumn and sensitize a largernumber of patients; Urticaceae reach thehighest pollen concentrations for a longerperiod but are not the primary cause ofpollinosis. This study shows that Z. fabagopollen becomes airborne, elicits an IgEresponse and, like other pollens, contributestowards triggering allergic symptoms.It should therefore be considered arelevant allergen and accordingly beincluded in skin test procedures.     

Seasonal variation in food allergy to apple

The aim of the study was to investigate the possibility of a seasonal variation in reactivity to apples in 27 birch pollen allergic patients. Before and during the birch pollen season 1998, the patients were subjected to double-blind, placebo-controlled food challenges (DBPCFCs) with grated fresh Golden Delicious apple followed by an open food challenge with whole fresh apple. The clinical reactions elicited during the challenges were evaluated both by the patients and the investigators. Moreover, the skin reactivity and the in vitro reactivity to apple were evaluated by skin prick test (SPT), leukocyte histamine release (HR), measurement of specific IgE, and immunoblotting experiments. The sensitivity of the DBPCFC, when compared with the result of the open challenge, was 0.74 (14/19) before the season and 0.80 (16/20) during the season. None of the patients reacted to the blinded challenge without a subsequent reaction to the open challenge. One placebo reaction was registered both before and in season, but not in the same patient. The patient scores of the first positive challenges, and the maximal scores of each combined blinded and open challenge session, were significantly increased during the pollen season (P<0.05). The scores of the open challenge were significantly higher than the scores of the DBPCFC both before the season and during the in-season challenges (P<0.05). Specific IgE against Golden Delicious increased during season (P<0.05), while neither SPT, HR, nor immunoblotting experiments could confirm an increase in reactivity. In conclusion, the results of the oral challenge tests indicated an increase in clinical reactivity to apples during the birch pollen season in birch pollen allergic individuals.     

Prevalence of intestinal parasites among individuals with allergic skin diseases

The prevalence of intestinal protozoans and helminths in stool samples of individuals with allergic cutaneous symptoms was evaluated to study a possible link between parasites and allergy. Altogether, 218 patients who had chronic urticaria, atopic dermatitis, or pruritus of unknown origin were included in the study. Standard laboratory tests for the detection of allergic etiology were performed for all patients. The presence of intestinal parasites was investigated using microscopy, immunofluorescence, and immunoenzymatic assays. Overall, protozoans and helminths were recovered from the stools of 48 subjects (P = 0.004), 18 of whom were affected with intestinal symptoms (P = 0.023). The presence of Giardia lamblia in the stools was significantly associated with allergic cutaneous manifestations (P = 0.030). In addition, patients with allergy were significantly more likely to have > or = 5 Blastocystis hominis organisms per field (P = 0.046). There was a set of patients with allergic cutaneous diseases in whom the presence of intestinal parasites may not be incidental.     

Pollinosis in trentino (Northern Italy). Aerobiological and clinical research

Summary A study to evaluate and define the atmospheric pollen concentration in Trentino was carried out through the aerobiologic sampling in three localities chosen according to their different climatic conditions.1375 patients with pollinosis living in Trentino were studied retrospectively over the period ranging from 1986 to 1988 and selected according to the area they came from.Results have proved that the most allergenic pollen types are the following: Poaceae, Urticaceae (Parietaria), Compositae (Artemisia) and the tree pollen of Betulaceae and Corylaceae (Alnus, Betula, Corylus), and that pollinosis caused by such pollen, types has different features and frequencies according to the different localities.As far as symptoms are concerned, our data shows that rhinoconjuntivitis is more frequent in those patients who are allergic toParietaria while asthma results being more frequent in patients who are allergic to tree pollen.