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Molecular and immunological characterization of a novel timothy grass (Phleum pratense) pollen allergen, Phl p 11.

BACKGROUND: Allergy to grass pollen is typically associated with serum IgE antibodies to group 1 and/or group 5 allergens, and additionally often to one or several less prominent allergens. Most of the grass pollen allergens identified to date have been characterized in detail by molecular, biochemical and immunological methods, timothy grass being one of the most thoroughly studied species. However, a 20-kDa allergen frequently recognized by IgE antibodies from grass pollen allergics has so far escaped cloning and molecular characterization. OBJECTIVE: To clone and characterize the 20 kDa timothy grass pollen allergen Phl p 11. METHODS: Phl p 11 cDNA was cloned by PCR techniques, utilizing N-terminal amino acid sequence obtained from the natural allergen. Phl p 11 was expressed as a soluble fusion protein in Escherichia coli, purified to homogeneity and used for serological analysis and to study Phl p 11 specific induction of histamine release from basophils and skin reactivity in sensitized and control subjects. RESULTS: Phl p 11 cDNA defined an acidic polypeptide of 15.8 kDa with homology to pollen proteins from a variety of plant species and to soybean trypsin inhibitor. The sequence contained one potential site for N-linked glycosylation. Serological analysis revealed that recombinant Phl p 11 shared epitopes for human IgE antibodies with the natural protein and bound serum IgE from 32% of grass pollen-sensitized subjects (n = 184). Purified recombinant Phl p 11 elicited skin reactions and dose-dependent histamine release from basophils of sensitized subjects, but not in non-allergic controls. CONCLUSION: As the first representative of group 11 grass pollen allergens, Phl p 11 has been cloned and produced as a recombinant protein showing allergenic activity. One-third of grass pollen-sensitized subjects showed specific IgE reactivity to recombinant Phl p 11, corresponding in magnitude to a significant proportion of specific IgE to grass pollen extract.

Amino Acid Sequence↗

Grass group I allergens (beta-expansins) are novel, papain-related proteinases.

Expansins are a family of proteins that catalyse long-term extension of isolated plant cell walls due to an as yet unknown biochemical mechanism. They are divided into two groups, the alpha-expansins and beta-expansins, the latter group consisting of grass group I allergens and their vegetative homologs. These grass group I allergens, to which more than 95% of patients allergic to grass pollen possess IgE antibodies, are highly immunologically crossreactive glycoproteins exclusively expressed in pollen of all grasses. Alignments of the amino-acid sequences of grass group I allergens derived from diverse grass species reveal up to 95% homology. It is therefore likely that these molecules share a similar biological function. The major grass group I allergen from timothy grass (Phleum pratense), Phl p 1, was chosen as a model glycoprotein and expressed in the methylotrophic yeast Pichia pastoris to obtain a post-translationally modified and functionally active allergen. The recombinant allergen exhibited proteolytic activity when assayed with various test systems and substrates, which was also subsequently demonstrated with the natural protein, nPhl p 1. These observations are confirmed by amino-acid alignments of Phl p 1 with three functionally important sequence motifs surrounding the active-site amino acids of the C1 (papain-like) family of cysteine proteinases. Moreover, the significantly homologous alpha-expansins mostly share the functionally important C1 sequence motifs. This leads us to propose a C1 cysteine proteinase function for grass group I allergens, which may mediate plant cell wall growth and possibly contributes to the allergenicity of the molecule.

Allergens↗

Competition for nitrogen between Australian native grasses and the introduced weed Nassella trichotoma.

BACKGROUND: and Aims Nassella trichotoma is an unpalatable perennial grass weed that invades disturbed native grasslands in temperate regions of south-eastern Australia. This experiment investigated whether elevated N levels, often associated with disturbance, increases the competitiveness of N. trichotoma relative to C3 and C4 native Australian grasses. METHODS: A pot experiment investigated competitive interactions between four native grasses, two C3 species (Microlaena stipoides and Austrodanthonia racemosa) and two C4 species (Themeda australis and Bothriochloa macra), and N. trichotoma at three different N levels (equivalent to 0, 60 and 120 kg ha-1) and three competing densities (zero, one and eight neighbouring plants), using an additive design. KEY RESULTS: All native grasses were competitive with N. trichotoma at low N levels, but only M. stipoides was competitive at high N. High densities of native grasses (8:1) had a major competitive effect on N. trichotoma at all N levels. The competitive ranking of native grasses, across all N levels, on N. trichotoma was: M. stipoides>A. racemosa>B. macra>T. australis. The C3 species were generally more competitive than the C4 species and C4 grasses were not inherently more productive at low N levels, in contrast to the results of other studies. CONCLUSION: To resist invasion from N. trichotoma, these native grasses need to be maintained at a high density and/or biomass. The results do not support the theory that species such as N. trichotoma, with high tissues density, are always less competitive than those of low tissue density; in this case competitiveness depended on N levels. The ability of N. trichotoma to accumulate biomass at a higher rate than these native grasses, helps to explain why it is a major weed in disturbed Australian native grasslands.

Australia↗

Specific cellular and humoral immunity in children with grass pollen asthma.

Lymphocyte stimulation, as determined by incorporation of thymidine, to rye grass extract in twenty-three children with bronchial reactivity to rye grass and to house dust mite, did not differ significantly from four children with reactivity to house dust mite alone, or from nine children with asthma but without a bronchial response to these allergens. Sixteen children underwent hyposensitization with rye grass extract or treatment with placebo. There was no consistent effect of hyposensitization on the lymphocyte stimulation indices to rye grass. A decrease in lymphocyte responsiveness occurred to rye grass and to house dust mite after the grass pollen season but was not statistically significant. Analysis of changes in lymphocyte responsiveness to both house dust mite and rye grass of the children most highly sensitized to rye allergen, showed that the lymphocyte responsiveness to rye grass fell during the pollen season (P less than 0.05) but this effect was not seen with house dust mite. The study suggests that a decrease in lymphocyte responsiveness to rye grass allergen in children with large amounts of anti-rye IgE antibodies is antigen specific and may be seen following seasonal exposure.

Animals↗

Comparison of IgE and IgG antibody responses of atopic individuals with sensitization to tree and grass pollens.

Sera of atopic individuals with predominant sensitization to either tree pollen (TAs) or tree and grass pollens (TGAs) as well as of nonatopic subjects (NAs) were analyzed for IgE, IgG, and IgG4 antibodies specific for grass pollens allergens. Of 600 atopic individuals with serum IgE antibodies specific for birch pollen allergens, 54% also had serum IgE antibodies specific for grass pollen. The mean titers of IgG antibodies specific for grass pollen proteins were about 10 times higher in the sera of TGAs than those in the TAs and NAs. SDS-PAGE immunoblotting analysis of grass pollen proteins using sera of TGAs, TAs, and NAs with respect to the binding of these proteins with IgE and IgG antibodies in these sera exhibited a similar pattern of variation. Quantitation by enzyme immunoassay of the antibody binding to a recombination grass pollen allergen, rKBG8.3, further demonstrated the elevated IgG antibody levels in TGAs are mainly due to a broader range of specificities, and not to high specific binding to the individual protein. Statistically significant correlation was found between IgE and IgG4 antibodies specific for the Kentucky bluegrass (KBG) extract, but not for the isolated recombinant allergen. These results indicate that the grass pollens elicit a complex array of antibody specificities in both atopics and nonatopics, and that the profile of antibodies specific to the pollen extract and pure allergens differs, suggesting that single grass allergens may be inadequate for replacing grass pollen extracts for immunotherapy.

Allergens↗

Antibody profiles and self-reported symptoms to pollen-related food allergens in grass pollen-allergic patients from northern Europe.

BACKGROUND: Most studies on pollen-related food allergy have so far focused on the association of birch/weed pollen allergens and plant food allergy. The aim of this study was to elucidate the allergen spectrum among a group of grass pollen-allergic patients from northern Europe and to relate the results to clinical histories of pollen-related food allergy. METHODS: Fifty-eight grass pollen-allergic patients answered a questionnaire regarding allergy to foods. Blood samples were taken to test IgE-reactivity to a large panel of pollen allergens and pollen- and nonpollen-related food allergens using crude allergen extracts and recombinant and native allergens. RESULTS: Three different groups of grass pollen-allergic patients were identified according to their IgE antibody profile: a grass pollen group only (19%), a grass and tree pollen group (29%) and a grass, tree and compositae (pan-) pollen group (48%). No sensitization to Bet v 1 as well as almost no IgE to plant food was observed in the grass pollen group. In contrast, nearly all patients in the two tree-related groups had IgE to Bet v 1, which reflected the high frequency of adverse reactions to typical birch-related food in these groups. Only four patients belonging to the pan-pollen group displayed IgE to profilin Phl p 12/Bet v 2. Patients in the pan-pollen group reported significantly more symptoms to food allergens compared with patients in the two other groups. The most frequently reported symptom was the oral allergy syndrome. CONCLUSIONS: Sensitization to grass pollen alone is rare among grass pollen-allergic patients from northern Europe. The majority of patients are in addition sensitized to birch (Bet v 1), which seems to be closely related to their pollen-derived food allergy. The study highlights the advantage of using well-defined allergen molecules for the diagnosis of cross-reactivity between pollen and food allergens.

Adult↗

Use of tall fescue EST-SSR markers in phylogenetic analysis of cool-season forage grasses.

Microsatellites or simple sequence repeats (SSRs) are highly useful molecular markers for plant improvement. Expressed sequence tag (EST)-SSR markers have a higher rate of transferability across species than genomic SSR markers and are thus well suited for application in cross-species phylogenetic studies. Our objectives were to examine the amplification of tall fescue EST-SSR markers in 12 grass species representing 8 genera of 4 tribes from 2 subfamilies of Poaceae and the applicability of these markers for phylogenetic analysis of grass species. About 43% of the 145 EST-SSR primer pairs produced PCR bands in all 12 grass species and had high levels of polymorphism in all forage grasses studied. Thus, these markers will be useful in a variety of forage grass species, including the ones tested in this study. SSR marker data were useful in grouping genotypes within each species. Lolium temulentum, a potential model species for cool-season forage grasses, showed a close relation with the major Festuca-Lolium species in the study. Tall wheat grass was found to be closely related to hexaploid wheat, thereby confirming the known taxonomic relations between these species. While clustering of closely related species was found, the effectiveness of such data in evaluating distantly related species needs further investigations. The phylogenetic trees based on DNA sequences of selected SSR bands were in agreement with the phylogenetic relations based on length polymorphism of SSRs markers. Tall fescue EST-SSR markers depicted phylogenetic relations among a wide range of cool-season forage grass species and thus are an important resource for researchers working with such grass species.

Alleles↗

Using plant breeding and genetics to overcome the incidence of grass tetany.

Plant breeders developing cultivars to minimize the hazards of grass tetany are concentrating largely on increasing herbage Mg concentrations in cool-season (C3) grasses. Significant genetic variation has been found for Mg, Ca and K concentrations within C3 grass species studied to date. For most C3 forage grass species, heritability estimates are highest for Mg, slightly lower for Ca and lowest for K concentrations. The largest genotype x environmental interactions are found for K values, whereas small environmental effects have been observed for Mg and Ca values. No C3 forage grass cultivar has been developed to date that would eliminate hypomagnesemia. Grass breeders need to develop more experimental C3 plant populations that have high Mg and Ca concentrations. These experimental synthetics with genetically altered mineral concentrations need to be fed to ruminants susceptible to grass tetany to determine whether grass tetany can be eliminated or reduced. Limited feeding trials using ruminants show that improved animal performance can be expected when feeding forage grasses bred for higher Mg concentrations.

Animals↗

Antigenic and allergenic relationships among airborne grass pollens in India.

BACKGROUND: Pollen from grasses (Poaceae) are predominant aeroallergens throughout the world including tropical countries. Studies from USA, Europe, and Australia have shown extensive allergenic/antigenic cross reactivity among the grass pollen allergens prevalent there. No such information is available about airborne grass pollens of tropical countries. OBJECTIVE: The present study was undertaken to explore common antigenic/allergenic components, if any, of five important grass pollens of India. METHODS: Intradermal tests (ID) were performed with pollen extracts of Cenchrus, Cynodon, Imperata, Pennisetum, and Sorghum in patients with nasobronchial allergy. ELISAs were performed for estimating the allergen-specific IgE in sera of patients eliciting markedly positive ID response (2+ to 4+). To detect cross reactivity, ELISA inhibition experiments were carried out using pooled patient sera and five grasses, individually, as inhibitors with different solid phase antigens. To evaluate common antigenic components in Cenchrus, Imperata, and Pennisetum pollen extracts, rocket immunoelectrophoresis (RIE) and ELISA inhibition were conducted using rabbit antisera. RESULTS: Among 133 patients, Cynodon extract elicited markedly positive skin reactivity in most patients followed by Pennisetum, Imperata, Cenchrus, and Sorghum. A large number of patients showed markedly positive skin reactions and enhanced specific IgE levels to more than one grass pollen extract. ELISA inhibition experiments showed different degrees of cross reactivity among the grass pollens studied. Rocket immunoelectrophoresis and ELISA inhibition using rabbit antisera with homologous and heterologous pollen revealed the presence of shared antigenic components in Cenchrus, Imperata, and Pennisetum extracts. CONCLUSION: The varied dose-response curves obtained with ELISA inhibition using different inhibitors suggest the presence of both common and specific antigens/allergens in the grass pollens studied. Based on the extensive immunologic cross reactivity, among the tropical grass species, it may be possible to use mixed allergen preparations for allergy diagnosis and immunotherapy.

Allergens↗

Comparison of natural and recombinant isoforms of grass pollen allergens.

More than 95% of grass pollen allergic patients possess IgE antibodies against grass group I, a heterogeneous group of glycoproteins found in all temperate grasses. We studied the structural variability of the group I allergens in single species and among different grasses. By 2-DE blotting using patients' IgE and monoclonal antibodies, we detected IgE-reactive isoforms with molecular masses between 32 and 37 kDa and focusing in a wide pI ranging from 4.7 to 7.6. While the group I allergens of timothy grass (Phl p 1) were composed of 37 and 35 kDa components, only single isoforms were found for ryegrass (Lol p 1) and velvet grass (Hol l 1): 32 and 34 kDa, respectively. By N-terminal microsequencing we determined single amino acid substitutions in different-sized group I allergens. The post-translational modifications (one N-glycosylation site, two hydroxylated proline residues and seven cysteine residues for potential disulfide formations), which contribute to IgE reactivity, were identical in all. From the cDNA sequences we deduced protein sequence homologies > 90%, a result which might explain the high IgE cross-reactivity among the grasses. In order to test whether recombinant group I grass allergens can act as substitutes for the natural forms, we expressed rPhl p 1 in E. coli and in P. pasteuris. 2-DE immunoblotting again demonstrated a microheterogeneity in molecular mass and pI. While the E. coli products were free from post-translational modifications, rPhl p 1 from Pichia is a heterogeneous glycoprotein fraction with a carbohydrate content of about 15%. This rPhl p 1 is hyperglycosylated compared to the nPhl p 1, which only has a 5% carbohydrate content.

Allergens↗

Influence of landfill gas on the microdistribution of grass establishment through natural colonization.

Many revegetated landfills have poor cover including bare areas where plants do not grow. This study, on the Bisasar Road Landfill site in South Africa, assessed grass species preferences to microhabitat conditions in a mosaic of patches of well-established grassed areas and bare, nonvegetated areas. Factors, including soil CO2, CH4, O2, nutrients, and other general soil conditions, were measured in relation to species distribution and grass biomass in the field. Cynodon dactylon was the dominant grass in the established grass areas but was less abundant in the areas bordering the bare areas where Paspalum paspalodes and Sporobolus africanus were common. A number of soil factors measured were significantly correlated with grass biomass and these included Mg, Ca, Zn, Mn, K, temperature, moisture, and CO2. However, a laboratory bioassay using the growth of C. dactylon with soils removed from the landfill indicated that there were no differences in the soils from the bare areas and those that supported high plant biomass. Thus, no nutrient deficiency or chemical toxicity was inherent in the soil in the laboratory. The results of the field investigation and bioassay indicated that soil CO2 as a result of landfill gas infiltration into the root zone was probably the main factor causing bare areas on the landfill where no grass species could colonize and grow and that C. dactylon was more sensitive to elevated soil CO2 than other grass species such as P. paspalodes and S. africanus.

Carbon Dioxide↗

Short photoperiod inhibits winter growth of subtropical grasses.

Grass development is influenced by length of photoperiod, but no direct measurements under natural conditions exist on mass accumulation in response to photoperiod by subtropical grass species. Grasslands of the subtropics are a major resource, but their growth is inhibited substantially during the short-photoperiod months. This research was designed to examine the consequences on grass production under field conditions when the limitation of short photoperiod is artificially removed. Lights, which extended the daylength to 15 h, were placed over plots of four subtropical forage grasses representing three species (Paspalum notatum Flugge; Cynodon dactylon L.; Cynodon nlemfuensis Vanderyst) to measure their mass accumulation in response to extended photoperiod in a 2-year experiment. Forage yields in all grasses at 5-week harvests during the time of shortest daylength were increased up to 6.2-fold more than the yield under the natural daylength. For the 4.5-month period of shortest daylength in each year, forage yields were increased for all grasses with one grass having a yield increase of 3.6-fold under the extended photoperiod as compared to natural daylength. These results demonstrated that selection of grasses that are insensitive to photoperiod could substantially increase forage yield of subtropical grasslands to benefit animal production and enhance carbon sequestration.

Light↗

A 30-day-ahead forecast model for grass pollen in north London, United Kingdom.

A 30-day-ahead forecast method has been developed for grass pollen in north London. The total period of the grass pollen season is covered by eight multiple regression models, each covering a 10-day period running consecutively from 21 May to 8 August. This means that three models were used for each 30-day forecast. The forecast models were produced using grass pollen and environmental data from 1961 to 1999 and tested on data from 2000 and 2002. Model accuracy was judged in two ways: the number of times the forecast model was able to successfully predict the severity (relative to the 1961-1999 dataset as a whole) of grass pollen counts in each of the eight forecast periods on a scale of 1 to 4; the number of times the forecast model was able to predict whether grass pollen counts were higher or lower than the mean. The models achieved 62.5% accuracy in both assessment years when predicting the relative severity of grass pollen counts on a scale of 1 to 4, which equates to six of the eight 10-day periods being forecast correctly. The models attained 87.5% and 100% accuracy in 2000 and 2002, respectively, when predicting whether grass pollen counts would be higher or lower than the mean. Attempting to predict pollen counts during distinct 10-day periods throughout the grass pollen season is a novel approach. The models also employed original methodology in the use of winter averages of the North Atlantic Oscillation to forecast 10-day means of allergenic pollen counts.

Climate↗

Acute phase proteins in grass sickness (equine dysautonomia).

Four acute phase proteins were assayed in the serum of normal horses and those with acute, subacute and chronic grass sickness, colic and inflammatory conditions, in order to investigate their diagnostic value in grass sickness. The grass sickness and inflammation group had a significantly increased haptoglobin concentration (P less than 0.01-P less than 0.001). Orosomucoid was elevated in acute, subacute and chronic grass sickness and inflammation (P less than 0.001, P less than 0.001, P less than 0.05 and P less than 0.05, respectively). Highest concentrations of haptoglobin and orosomucoid were recorded in subacute grass sickness. Ceruloplasmin was significantly higher in acute grass sickness cases than all other groups except the colic group (P less than 0.05-P less than 0.01). alpha 2-macroglobulin was significantly higher in acute grass sickness than normal, colic and chronic grass sickness cases (P less than 0.01, P less than 0.05 and P less than 0.05). The time scale of changes suggests that the stimulus to haptoglobin and orosomucoid synthesis occurs at the onset of clinical signs whereas the increase in ceruloplasmin and alpha 2-macroglobulin is more likely to reflect haemoconcentration.

Acute Disease↗

Grazers and grass: Monte Carlo simulations.

To illustrate the interplay between grazers and grass, we present a novel Monte Carlo model including grass-island growth, consumption of grass by grazers, and birth, migration and death of grazers. The rates of the former and three latter processes are assumed to depend on the environment so that the conventional mean-field approximation does not hold (in particular, the model takes into account that grass grows on the grass-island boundaries, and grazers are mobile and prefer to stay on the areas covered by grass). Due to the feedback between various processes, as expected, the model predicts stable regimes and irregular oscillations of the area of the grass islands and grazer population. The patterns observed are however different compared to those predicted by conventional Monte Carlo prey-predator models. Specifically, there is no tendency for grazers and grass to segregate. The mean-field version of the model is briefly discussed as well.

Algorithms↗

A molecular identification system for grasses: a novel technology for forensic botany.

Our present inability to rapidly, accurately and cost-effectively identify trace botanical evidence remains the major impediment to the routine application of forensic botany. Grasses are amongst the most likely plant species encountered as forensic trace evidence and have the potential to provide links between crime scenes and individuals or other vital crime scene information. We are designing a molecular DNA-based identification system for grasses consisting of several PCR assays that, like a traditional morphological taxonomic key, provide criteria that progressively identify an unknown grass sample to a given taxonomic rank. In a prior study of DNA sequences across 20 phylogenetically representative grass species, we identified a series of potentially informative indels in the grass mitochondrial genome. In this study we designed and tested five PCR assays spanning these indels and assessed the feasibility of these assays to aid identification of unknown grass samples. We confirmed that for our control set of 20 samples, on which the design of the PCR assays was based, the five primer combinations produced the expected results. Using these PCR assays in a 'blind test', we were able to identify 25 unknown grass samples with some restrictions. Species belonging to genera represented in our control set were all correctly identified to genus with one exception. Similarly, genera belonging to tribes in the control set were correctly identified to the tribal level. Finally, for those samples for which neither the tribal or genus specific PCR assays were designed, we could confidently exclude these samples from belonging to certain tribes and genera. The results confirmed the utility of the PCR assays and the feasibility of developing a robust full-scale usable grass identification system for forensic purposes.

Botany↗

Sublingual immunotherapy with once-daily grass allergen tablets: a randomized controlled trial in seasonal allergic rhinoconjunctivitis.

BACKGROUND: Specific immunotherapy is the only treatment modality that has the potential to alter the natural course of allergic diseases. Sublingual immunotherapy has been developed to facilitate access to this form of treatment and to minimize serious adverse events. OBJECTIVE: To investigate the efficacy and safety of sublingual grass allergen tablets in seasonal allergic rhinoconjunctivitis. METHODS: A multinational, multicenter, randomized, placebo-controlled trial conducted during 2002 and 2003. Fifty-five centers in 8 countries included 855 participants age 18 to 65 years who gave a history of grass pollen-induced allergic rhinoconjunctivitis and had a positive skin prick test and elevated serum allergen-specific IgE to Phleum pratense. Participants were randomized to 2500, 25,000, or 75,000 SQ-T grass allergen tablets (GRAZAX; ALK-Abelló, Hørsholm, Denmark) or placebo for sublingual administration once daily. Mean duration of treatment was 18 weeks. RESULTS: Average rhinoconjunctivitis scores during the season showed moderate reductions of symptoms (16%) and medication use (28%) for the grass allergen tablet 75,000 SQ-T (P = .0710; P = .0470) compared with placebo. Significantly better rhinoconjunctivitis quality of life scores (P = .006) and an increased number of well days (P = .041) were also observed. Efficacy was increased in the subgroup of patients who completed the recommended preseasonal treatment of at least 8 weeks before the grass pollen season (symptoms, 21%, P = .0020; and medication use, 29%, P = .0120). No safety concerns were observed. CONCLUSION: This study confirms dose-dependent efficacy of the grass allergen tablet. Although further studies are required, the greater tolerability of the tablet may permit immunotherapy to be available to a much broader group of patients with impaired quality of life caused by grass pollen allergy. CLINICAL IMPLICATIONS: For patients with grass pollen allergy, sublingual immunotherapy is well tolerated and can reduce symptoms and improve quality of life.

Administration, Sublingual↗

Immunologic cross-reactivity among cereal grains and grasses in children with food hypersensitivity.

BACKGROUND: Because of a lack of clinical data from food challenges, allergists often recommend dietary restriction of all cereal grains in patients with sensitivity to at least one grain. OBJECTIVES: The purposes of this study were to assess the degree of intrabotanical cross-reactivity among cereal grains and related grasses, to better define the prevalence of multiple grain hypersensitivity, and to define the protein fractions associated with wheat hypersensitivity. METHODS: One hundred forty-five patients evaluated by food challenges and skin prick tests were divided into three groups: group 1, cereal grain and grass allergies; group 2, wheat allergy alone; and group 3, grass allergy alone. Fifteen patients were further selected from groups 1 to 3. Sodium dodecylsulfate--polyacrylamide gel electrophoresis and immunoblot analyses were performed on six grains and four related grasses with sera from these patients. RESULTS: Only 21% of patients had symptomatic reactivity as determined by food challenge; 80% had reactivity to only one grain. As determined by immunoblot analyses, patients in groups 1 and 2 showed extensive cross-reactivity (within each group) among grains but little cross-reactivity among grasses, whereas patients in group 3 showed cross-reactivity between the grains and grasses. Patients with wheat allergy had specific IgE binding to wheat fractions 47 kd and 20 kd, bands not recognized by patients with grass allergy. CONCLUSIONS: Clinically insignificant cross-reactivity exists among cereal grains and grasses; therefore, elimination of all grains from the diet of a patient with grain allergy is unwarranted. Further purification and characterization of the 47 kd and 20 kd wheat fractions is needed to provide more specific in vitro testing.

Adolescent↗