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PubMed1992

Recombinant pollen allergens from Dactylis glomerata: preliminary evidence that human IgE cross-reactivity between Dac g II and Lol p I/II is increased following grass pollen immunotherapy.

We previously described the isolation of three identical complementary DNA (cDNA) clones, constructed from Orchard/Cocksfoot grass (Dactylis glomerata) anther messenger RNA (mRNA), expressing a 140,000 MW beta-galactosidase fusion protein recognized by IgE antibodies in atopic sera. Partial nucleotide sequencing and inferred amino acid sequence showed greater than 90% homology with the group II allergen from Lolium perenne (Lol II) indicating they encode the group II equivalent, Dac g II. Western blot immunoprobing of recombinant lysates with rabbit polyclonal, mouse monoclonal and human polyclonal antisera demonstrates immunological identity between recombinant Dac g II, Lol p I and Lol p II. Similar cross-identity is observed with pollen extracts from three other grass species: Festuca rubra, Phleum pratense and Anthoxanthum odoratum. Recombinant Dac g II was recognized by species- and group-cross-reactive human IgE antibodies in 33% (4/12) of sera randomly selected from grass-sensitive individuals and in 67% (14/21) of sera from patients receiving grass pollen immunotherapy, whilst 0/4 sera from patients receiving venom immunotherapy alone contained Dac g II cross-reactive IgE. Cross-reactive IgG4 antibodies were detectable in 95% of sera from grass pollen immunotherapy patients. These preliminary data suggest that conventional grass pollen allergoid desensitization immunotherapy may induce IgE responses to a cross-reactive epitope(s) co-expressed by grass pollen groups I and II (and possibly group III) allergens.

Allergens↗
PubMed1988

Study of the epitope structure of purified Dac G I and Lol p I, the major allergens of Dactylis glomerata and Lolium perenne pollens, using monoclonal antibodies.

The use of mAb allowed us to further analyze the cross-reactivity between purified Dac g I and Lol p I, the major allergens of Dactylis glomerata (cocksfoot) and Lolium perenne (Rye grass), respectively. It was first shown, using IEF, followed by immunoprinting, that serum IgE antibodies from most grass-sensitive patients recognize both Dac g I and Lol p I. Second, three different anti-Lol p I mAb, 290A-167, 348A-6, and 539A-6, and one anti-Dac g I mAb, P3B2 were all shown to react with Dac g I and Lol p I, indicating that the two molecules share common epitopes. Epitope specificity of the mAb was determined by competitive binding inhibition of a given labeled mAb to solid phase fixed Dac g I or Lol p I by the mAb. The results indicated that the four mAb are directed against four different and non-overlapping epitopes present on both allergens. Using double-binding RIA, our data strongly suggest that the common epitopes are not repetitive on both molecules. In addition to their similar physicochemical characteristics, such as isolectric points and m.w., Dac g I and Lol p I share four identical epitopes. Binding inhibition of human IgE to Lol p I and Dac g I by the mAb was also assessed. The results indicated that each mAb was able to inhibit such reactions to variable degree but no additive inhibition was observed when two mAb of different specificities were used in combination, suggesting that the human IgE binding site is partially shared by each epitope recognized by the four mAb.

Allergens↗
PubMed1983

A report on the consumption, composition and nutritional adequacy of a mixture of lush green perennial ryegrass (Lolium perenne) and cocksfoot (Dactylis glomerata) fed ad libitum to Thoroughbred mares.

Non-pregnant Thoroughbred mares were stabled and subjected to 2 trials, each 24 h in duration, to establish their total consumption of a mixture of freshly cut, lush green perennial ryegrass (Lolium perenne) and cocksfoot (Dactylis glomerata) in approximately equal proportions; and to compare the total intake of crude protein, calcium, phosphorus and mass of the grass mixture on a dry matter basis with their daily nutritional requirements. The body mass of each mare was calculated at the commencement of each trial. In the first trial 2 lactating mares with foals at foot, 65 days and 8 days of age, and one mature non-lactating mare, consumed 75,5 kg, 61,0 kg and 39,5 kg of the grass mixture, cut in the early vegetative stage, respectively. The perennial ryegrasss (L. perenne) contained 79,63% moisture, 1,67% crude protein, 0,75% calcium, 0.057% phosphorus and 20.37% dry matter. The cocksfoot (D. glomerata) contained 79,52% moisture, 2,27% crude protein, 0,051% calcium, 0,061% phosphorus and 20,48% dry matter. The younger foal did not eat the grass mixture. The mass of grass mixture consumed by the older foal was not determined. In the second trial conducted 3 weeks later, when oat straw (Avena sativa) was also fed ad lib itum, the same lactating mares, but a different mature non-lactating mare, consumed 54.5 kg, 56.0 kg and 40.5 kg of the grass mixture, cut in the mid bloom stage, respectively. The perennial ryegrass (L. perenne) contained 70,03% moisture, 2,37% crude protein, 0,086% calcium 0,068% phosphorus and 29,97 dry matter.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗
PubMedDate not supplied

[The monitoring of natural populations of Dactylis glomerata L. in the area of the accident at the Chernobyl Atomic Electric Power Station].

Results of five years monitoring (1986-1991) of seed quality (SQ) and chlorophyll mutation frequency (MF) of Dactylis glomerata L., growing within 30-km zone of Chernobyl NPP are presented. It was shown that during initial postaccident period (1986, 1987) the criteria depend of the gamma-phone on the sampling plots. It was not found the correlation between SQ, MF and duration of plant growing under condition chronic irradiation.

Analysis of Variance↗
PubMed2006

Fractional and structural characterization of hemicelluloses from perennial ryegrass (Lolium perenne) and cocksfoot grass (Dactylis glomerata).

Sequential three-stage treatments with 80% EtOH containing 0.2% NaOH, 2.5% H2O2-0.2% EDTA containing 1.5% NaOH and 2.5% H2O2-0.2% TAED containing 1.0% NaOH at 75 degrees C for 3h released 8.0% and 10.4%, 79.1% and 77.0% and 12.9% and 12.5% of the original hemicelluloses from perennial grass and cocksfoot grass, respectively. It was found that the four alkaline peroxide-soluble hemicellulosic fractions contained higher amounts of xylose (33.4-38.2%), uronic acids (9.3-15.3%) and rhamnose (3.0-3.9%), but were lower in glucose (25.1-28.3%), galactose (13.3-15.3%) and mannose (0.4-1.5%) than those of the two alkaline EtOH-soluble hemicellulosic fractions in which glucose (32.9-36.0%), xylose (20.1-22.6%), arabinose (14.1-21.4%), galactose (16.6-19.9%), mannose (4.1-9.9%) and uronic acids (3.4-7.4%) were the major sugar components. 13C NMR spectroscopy confirmed that all the six hemicellulosic fractions were composed of galactoarabinoxylans, 4-O-methylglucuronoarabinoxylans and beta-glucan. In addition, the studies showed that the four alkaline peroxide-soluble hemicellulosic fractions were more linear and acidic and had larger molecular weights (Mw, 28,400-38,650 g mol(-1)) than those of the two alkaline EtOH-soluble hemicellulosic fractions (Mw, 16,460-17,420 g mol(-1)).

Chemical Fractionation↗
PubMed2005

The major grass pollen group 5 allergen from Dactylis glomerata and its C-terminal split product both behave as dimers: implications for allergen standardization.

BACKGROUND: On SDS-PAGE grass pollen group-5 allergens migrate as a doublet with an apparent molecular mass (M(r)) of 25 kDa. Immunoblot analysis revealed additional group 5 reactivity at double and half this M(r). The aim of this study was to investigate these group 5 molecular entities and to compare their allergenicity and behavior in quantitative immunoassays. METHODS: Group-5-specific monoclonal antibodies were produced and used for the development of a group-5-specific sandwich ELISA. Affinity-purified Dac g 5 was separated by SDS-PAGE/Western blotting; individual bands were analyzed by N-terminal sequencing. Size exclusion chromatography (SEC) in conjunction with group-5-specific ELISA, competitive RIA and RAST inhibition were used to analyze the size distribution of Dac g 5. Basophil histamine release assays were used to assess biological activity. RESULTS: The lower band of the typical group 5 doublet was identified as a truncated form lacking the typical group 5 N-terminus AD(L)/(A)GY, observed in the upper band. The 12-kDa peptide was shown to be the C-terminal half of Dac g 5 (amino acid 127 onwards). SEC in conjunction with competitive RIA revealed that around 45% of Dac g 5 is represented by the 12-kDa peptide. Both the C-terminal half and the whole allergen dimerize under nondenaturing conditions. In competitive RIA and RAST inhibition both forms are equally well detected. In contrast, the half molecule is poorly recognized in sandwich ELISA and displays negligible biological activity in basophil histamine release tests with purified IgE. CONCLUSIONS: These observations stress the need to evaluate the performance of allergen standardization protocols in detail, with special attention to allergen size distribution.

Allergens↗
PubMed1985

Identification of orchard grass (Dactylis glomerata) pollen allergens following electrophoretic transfer to nitrocellulose.

Orchard grass (cocksfoot) pollen extracts, fractionated by polyacrylamide gradient electrophoresis or SDS gel electrophoresis were electroblotted onto nitrocellulose membranes and probed with sera from orchard grass pollen-allergic patients and 125I-anti-human IgE. The IgE-binding components of the pollen were detected by autoradiography. Elution studies showed that allergens could be extracted immediately and continuously over a 3-hour period. Two fractions of MWs 28,000 and 30,000 could be detected only after 20 min extraction. SDS-PAGE separations gave the better resolution revealing 19 electrophoretically-separate components, 13 of which bound human IgE. All of the IgE-binding components had MWs in the range 14,000 - 70,000. Three of the bands bound IgE from more than 85% of the serum samples. Following gradient gel electrophoresis, IgE binding was exhibited by 10 bands in the range MW 5,000 to greater than 669,000. The technique used allows one to quantitatively examine patients' sera for allergen-specific IgE antibodies and to identify the clinically important allergens. Results revealed numerous allergenic components over a wide MW range while patterns of IgE binding with different patients' sera demonstrated a great diversity of IgE antibody responses. This study demonstrates the suitability of the electroblotting technique combined with autoradiography for the investigation of allergenic components of grass pollen extracts and hence has application to extract standardization and immunotherapy. Such studies can be carried out rapidly, economically and with a high degree of sensitivity.

Allergens↗
PubMedDate not supplied

[Obtaining mutations with cytoplasmic male sterility (CMS) in Dactylis glomerata L. under the influence of chemical mutagens].

When treating seeds of orchard grass with definite concentrations of nitrosoethylurea (NEU), dimethyl sulphate (DMS) and ethylene imine (EI) in M1 and M2 the plants with CMS as well as sterility fixators to them are obtained. Proportionality of mutagenic action and stimulation effect of these substances are found. The greatest amount of sterility fixators is found in M1 among plants the seeds of which were treated with NEU and EI; in M2 the fixators are found among plants obtained after treating seeds with DMS. Sterility fixators were found among plants of the VIK-61 and Asta and VIK-61XAsta hybrid population.

Aziridines↗