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PubMed2001

Measurement of IgE antibodies against purified grass-pollen allergens (Phl p 1, 2, 3, 4, 5, 6, 7, 11, and 12) in sera of patients allergic to grass pollen.

BACKGROUND: Current allergy diagnosis is performed with allergen extracts which contain a variety of allergenic and nonallergenic components. The availability of highly purified and well-characterized allergen molecules seems to be an advantage of component-based diagnosis. METHODS: With the immunoenzymatic CAP FEIA System, we measured specific IgE levels to the recombinant allergens rPhl p 1, rPhl p 2, rPhl p 5, rPhl p 6, rPhl p 7, rPhl p 11, rPhl p 12, and native Phl p 4 in 77 sera of patients allergic to grass pollen, in order to evaluate the IgE-binding frequency to these purified grass-pollen allergens and their relationship to rBet v 4, rBet v 2, and other allergens. RESULTS: The frequency of sensitization was as follows: rPhl p 1=93.5%; rPhl p 2=67.5%; rPhl p 5=72.7%; rPhl p 6=68.8%; rPhl p 7=7.8%; rPhl p 11=53.2%; rPhl p 12=35.1%; and native Phl p 4=88.3%. As expected, rPhl p 7 and rPhl p 12 had a very good correlation (Spearman's r) with Bet v 4 (r=0.95%, P<0.05) and rBet v 2 (r=0.99, P<0.05), respectively. Good correlations of rPhl p 12 with papain (r=0.93, P<0.05), latex (r=0.92, P<0.05), and bromelain (r=0.86, P<0.05) were found. Highly variable individual sensitization patterns were observed. CONCLUSIONS: A new clinical approach has allowed the determination of specific allergograms for the different patients and may therefore be of great importance for more specific diagnosis. The use of component-resolved diagnostics may be useful to evaluate the allergen content of an extract for immunotherapy by monitoring patient's IgE and IgG directed to relevant allergens.

Adolescent↗

PubMedDate not supplied

Canine tracheal mucus transport of particulate pollutants: comparison of radiolabeled corn pollen, ragweed pollen, asbestos, silica, and talc to Dowex anion exchange particles.

Canine tracheal mucous transport rates of each of two organic (corn and ragweed pollen), and three inorganic (chrysotile asbestos fibers, silica, and talc), 99mTc labeled pollutants were simultaneously measured against those of 125I labeled Dowex anion exchange resin particles in healthy animals. The method of labeling of the pollutants with 99mTc is described. With the exception of ragweed pollen, which moved 15% faster than the reference marker (P <.02), we found no difference in the transport rates of these particulate pollutants and the reference marker. We conclude that the particulate pollutants studied are handled by the mucociliary system in a similar fashion, and that the transport rates of Dowex particles represent valid measurements of the transport rates of these pollutants.

Animals↗

PubMed1978

A comparative study of an aqueous grass pollen extract and glutaraldehyde-treated grass pollen-tyrosine adsorbate in the treatment of pollenosis.

Hyposensitization therapy gave a subjective improvement in 84% of grass pollen-sensitive patients receiving an eighteen dose course of aqueous pollen extract (SDV) and in 69% of patients receiving a three dose course of glutaraldehyde-treated pollentyrosine adsorbate (Pollinex). Systemic reactions to treatment were considerably more frequent with the aqueous course, but virtually absent with the short adsorbate course, which seems therefore more convenient for the patient. The SDV treatment induced a more pronounced IgG antibody response both in patients and in guinea-pigs receiving the clinical dosage schedule. The Pollinex treatment showed a tendency to enhance the IgE antibody response in new patients.

Adult↗

PubMed2005

Pollen flow in the wildservice tree, Sorbus torminalis (L.) Crantz. II. Pollen dispersal and heterogeneity in mating success inferred from parent-offspring analysis.

Knowing the extent of gene movements from parents to offspring is essential to understand the potential of a species to adapt rapidly to a changing environment, and to design appropriate conservation strategies. In this study, we develop a nonlinear statistical model to jointly estimate the pollen dispersal kernel and the heterogeneity in fecundity among phenotypically or environmentally defined groups of males. This model uses genotype data from a sample of fruiting plants, a sample of seeds harvested on each of these plants, and all males within a circumscribed area. We apply this model to a scattered, entomophilous woody species, Sorbus torminalis (L.) Crantz, within a natural population covering more than 470 ha. We estimate a high heterogeneity in male fecundity among ecological groups, both due to phenotype (size of the trees and flowering intensity) and landscape factors (stand density within the neighbourhood). We also show that fat-tailed kernels are the most appropriate to depict the important abilities of long-distance pollen dispersal for this species. Finally, our results reveal that the spatial position of a male with respect to females affects as much its mating success as ecological determinants of male fecundity. Our study thus stresses the interest to account for the dispersal kernel when estimating heterogeneity in male fecundity, and reciprocally.

Conservation of Natural Resources↗

PubMed1975

Cutaneous reactions to pollen extracts in patients with hay fever. Special references to grass pollen recommended for treatment in fixed combination.

Skin reactivity to 8 common grass pollens was investigated in patients with hay fever using commercial, unfractionated extracts in serial dilution technique. The highest degree of sensitivity was observed for cocksfoot, meadow fescue, meadow foxtail, rye grass, timothy and Yorkshire fog. Five of those are included in the Allpyral 5-Grass Mix. The cutaneous responses to rye grass pollen overlap with those to timothy in a high degree.

Adolescent↗

PubMed2005

Cytokine and antibody responses in birch-pollen-allergic patients treated with genetically modified derivatives of the major birch pollen allergen Bet v 1.

BACKGROUND: Recently, recombinant hypoallergenic derivatives of the major birch pollen allergen, Bet v 1, were used to treat birch-pollen-allergic patients in a double-blind, placebo-controlled, multi-centre immunotherapy study. The aim of this study was to evaluate the effects of vaccination with aluminium-hydroxide-adsorbed recombinant Bet v 1 derivatives versus placebo on T-cell, cytokine and antibody responses in a subgroup of patients. METHODS: Blood was drawn from patients of the Swedish centre (n = 27; rBet v 1 fragments: n = 10; rBet v 1 trimer: n = 8, and placebo-aluminium hydroxide: n = 9) before the start and after completion of the treatment. PBMC were stimulated with rBet v 1 and analysed for cytokine (IL-4, IL-5, IL-10, IL-12, IL-13 and IFN-gamma)-secreting cells by ELISpot. Bet v 1-specific antibody levels in serum (IgG(1-4), IgE and IgA) were measured by ELISA. Skin prick tests with defined Bet v 1 concentrations were performed before and 10-11 months after the beginning of the study. RESULTS: Bet v 1-specific IgG levels, consisting of IgG(1), IgG(2) and IgG(4), were significantly increased after treatment with recombinant allergen derivatives. Treatment with rBet v 1 trimer led to a significant (p < 0.05) reduction of Bet v 1-reactive IL-5- and IL-13-producing cells, reflecting a reduced Th2 response. In addition, a decreased number of Bet v 1-reactive IL-4 producing (p = 0.07) and an increase of IL-12-producing (p = 0.06) cells was noted in the trimer-treated patients. In contrast to placebo, active treatment resulted in significantly reduced immediate-type skin reactions to Bet v 1 even 10-11 months after treatment. CONCLUSION: Vaccination with recombinant hypoallergenic Bet v 1 derivatives induces a Bet v 1-specific IgG response and leads to reduced skin reactivity in allergic patients. A reduction of Bet v 1-specific Th2 responses was observed in trimer-treated patients, which may reflect the intrinsic property of this allergen derivative.

Adult↗

PubMed1980

Allergens of Kentucky Blue Grass pollen. II. Isolation of hapten-like components from Kentucky Blue Grass pollen by preparative isoelectrofocussing.

Components with hapten-like properties were isolated from the nondialyzable fraction, i.e. the retentate (R) and the dialyzable fractions of the aqueous extract of Kentucky Blue Grass pollen (KBC aq.ext.), by preparative isoelectrofocussing on Sephadex G-100 gel. These haptenic components could not elicit the passive cutaneous anaphylaxis (PCA) reactions in rats passively sensitized with a murine reaginic antiserum to R, but could inhibit completely and specifically the PCA reaction which is normally elicitable with R. It was concluded that the specificity of the murine IgE antibodies was directed to a determinant(s) which was common to either allergenic or haptenic fractions. Moreover, by employing a pool of human sera from individuals allergic to KBG pollen in the radioallergosorbent test procedure, it was apparent that most of the haptenic fractions lacked some of the specificities present on allergenic components of R that were recognized by the human IgE antibodies. Evidence was obtained to suggest that the electrophoretic heterogeneity of allergenic components present in various fractions of KBG aq.ext. may be due primarily to differences in their net charge, rather than to differences in their allergenic specificity.

Absorption↗

PubMed1987

Immunogenicity in guinea pigs and tolerance in grass pollen-sensitive volunteers of enteric-coated grass pollen allergens.

An acid-insoluble, methacrylic acid, methyl methacrylate copolymer (Eudragit L-100) was used to give an enteric coating to a grass pollen extract in order to protect it against gastric degradation. Substantial protection against the degradative effects of simulated gastric secretion was demonstrated using this preparation which was well tolerated by grass pollen-allergic volunteers. The enteric-coated allergen induced a greater secondary antibody response than did an aqueous presentation when administered orally to guinea pigs which had been primed previously by subcutaneous injection. This result indicates that an effective hyposensitisation regimen could consist of a short series of initial parenteral injections, followed by an oral course of the protected allergen.

Acrylic Resins↗

PubMed1980

Characteristics of pollen diffusates and pollen wall cytochemistry in poplars.

Pollen diffusates, known to be important in pollen-stigma interactions controlling interspecific incompatibility between Populus deltoides and Populus alba, have been partly characterized and shown to contain more than 20 protein bands by polyacrylamide gel electrophoresis, at least 4 of these being glycoproteins. Seven fractions had antigenic activity in rabbits and several enzyme activities were also present. Peroxidase and leucine aminopeptidase isoenzymes were detected in the diffusates, demonstrating the extracellular location of these 2 enzymes. Isoenzyme patterns of peroxidase, esterase and acid phosphatase were complex, with some bands common to both species. Localization of acid phosphatase in the intine and esterase in the exine was demonstrated after brief aldehyde fixation and low-temperature embedding in glycol methacrylate. The intine and exine sites were distinguished by their chemical and structural features. Calcofluor white M2R new proved to be an excellent stain for differentiating the intine. Aniline blue-positive material, probably beta-1,3-glucan, is present associated with the intine of many ungerminated as well as germinating grains: production of this material may be a response to damage.

Antigens↗

PubMed1991

Airborne pollen and fungal spore sampling on the central California coast: the San Luis Obispo pollen project.

A semiarid coastal location in San Luis Obispo, California was surveyed for 3 years (1986-1988) using a Rotorod sampler. Significant year-to-year variations in predominant pollen occurred, and abundant levels of fungal spores were observed. Coincidently, a large wildlands fire that may have affected pollen levels occurred in the region shortly before sampling began. The entire survey period took place during a drought.

Air Pollutants↗

PubMed1989

Bahia grass pollen, a significant aeroallergen: evidence for the lack of clinical cross-reactivity with timothy grass pollen.

Thirty-eight subjects were challenged (25 nasal, 13 bronchial) with Bahia grass, Paspalum notatum, pollen extract. A positive Bahia intradermal skin test predicted a positive challenge to Bahia in all (11/11) of the nasal challenges and 75% (6/8) of the bronchial challenges. All 19 subjects with negative Bahia intradermal skin tests had negative challenges with Bahia. Specific IgE antibodies to Bahia pollen were detected by conventional RAST (greater than or equal to 2+) in 82% (14/17) of subjects with positive challenges and in 5% (1/20) of subjects with negative challenges. Eight subjects had positive intradermal skin tests to either Bahia (three) or timothy, Phleum pratense (five). Seven of the eight subjects reacted exclusively to either Bahia or timothy nasal challenge as predicted by their skin tests. Bahia grass is a significant aeroallergen, which in some subjects can be demonstrated not to cross-react with timothy.

Allergens↗

PubMed1986

Comparative studies on tree pollen allergens. XIII. Partial characterization of the alder (Alnus incana) pollen extract by two-dimensional IEF/SDS-PAG electrophoresis combined with electrophoretic transfer and immunoautoradiography.

Horizontal isoelectric focusing (IEF) and two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (2-D SDS-PAGE) combined with electrophoretic transfer (ET) and immunoautoradiography (IAR) were used to characterize the crude alder (Alnus incana) pollen extract (AI crude) and fraction AI 34. IEF resolved AI crude and AI 34 into 34 and 39 distinct protein bands, respectively, when stained with Coomassie brilliant blue. The band patterns were similar and the majority of bands were located in the pH region 4.0-7.0. After ET and IAR, the distribution of antigenic reactivities was similar to that of the protein stain. Allergenic reactivity was detected in the pH region 4.6-5.2 only, where two bands at pH 4.9 demonstrated dominating IgE-binding properties. The 2-D reference maps of AI crude and AI 34 consisted of 40 and 45 protein spots, respectively. The autoradiogram of AI crude demonstrated 35 spots capable of binding rabbit IgG. The main IgE-binding zone was located at pH 4.6-5.2 and 20 kilodaltons with an area of weaker autoradiographic signals at pH 4.6-5.2 and 40 kilodaltons. ET from polyacrylamide gels to nitrocellulose (NC) membranes was quantitative and IAR gave significant radiostaining of spots containing 0.01 and 0.1 microgram of AI crude in the IgG and IgE systems, respectively. For the characterization of the alder pollen extract, horizontal 2-D SDS-PAGE combined with ET and IAR provided a powerful supplement to conventional crossed immuno- and radioimmunoelectrophoretic techniques (CIE/CRIE).

Animals↗

PubMed1986

Allergens in pollen from mugwort (Artemisia vulgaris L.). II. Characterization of a crude and a partly purified extract of mugwort pollen with particular emphasis on the glycoprotein allergen Ag7.

A crude and a partly purified extract of mugwort pollen were characterized with particular emphasis on the glycoprotein allergen Ag7. Crossed immunoaffinoelectrophoresis with immobilized Con A in the intermediate gel demonstrated binding of Ag7 to this lectin, indicating that Ag7 contains terminal alpha-D-mannopyranosyl and/or alpha-D-glucopyranosyl residues. Crossed immunoaffinoelectrophoresis with free Con A in the first-dimension gel demonstrated microheterogeneity in the carbohydrate moiety of the allergen. Crossed radioimmunoelectrophoresis analysis using sera from 26 mugwort-allergic patients indicated that Ag7 is an important allergen in mugwort pollen. Heat stability experiments demonstrated the presence of at least three heat-resistant allergens, one of these being Ag7. Crossed immunoelectrofocusing indicated a pI value for Ag7 of 4.3.

Allergens↗

PubMed1983

Comparative studies on tree-pollen allergens. V. Immunochemical mapping of the antigens and allergens of birch pollen extract (Betula verrucosa).

A comparative electrophoretic and autoradiographic analysis of the different fractions of birch pollen crude extract was performed. This extract included a minimum of 17 reproducible and distinct antigens located in the gel filtration fractions BV2-BV4. Only one of these precipitates had the ability to bind IgE, as demonstrated by immunoelectrophoretic and autoradiographic techniques using several poly- and monospecific rabbit antibodies. The allergenicity of the different fractions was examined in vitro by RAST and RAST inhibition, and in vivo by passive cutaneous transfer and skin prick tests. The data suggested that Betula verrucosa pollen extract contains a group of isoallergens with related antigenicity but with variable molecular sizes. The findings have also confirmed the presence of identical antigenic properties of the previously isolated pI 5.18 and BV45 as deduced from their immunoelectrophoretic and autoradiographic studies.

Allergens↗

PubMed1983

Comparative studies on tree-pollen allergens. VI. The effect of thermal denaturation on the reactivity of birch pollen (Betula verrucosa) allergens.

The structural changes as due to thermal denaturation of antigens and allergens of birch pollen produced by gel filtration chromatography were studied by circular dichroism, RAST inhibition and immunoelectrophoretic techniques. Results of circular dichroism showed that both BV3 and BV4 contained 20-25% alfa-helix, while BV45 consists of beta-pleated sheet and random coil. Fractions BV3 and BV4 lost about 50% of their native tertiary structures when heated at 100 degrees C for 3 h. The number of antigenic lines found in the CIE plates for the heat-treated fractions was reduced as well. The CRIE preparations showed unchanged radiostaining for the heated BV3 fraction (correlated to the untreated BV3 fraction). For the BV4 fraction a diminished radiostaining was observed. A reduction of 8-fold and 4-fold of the IgE binding capacity was observed in RAST inhibition for fractions BV3 and BV4, respectively after heating. The purified fraction BV45 showed 50% decrease in CD absorption after heating to 100 degrees C for 1 h. The CIE pattern of the heated fraction gave a single precipitate line and the area under the precipitate line was reduced as compared to the unheated fraction. The CRIE plate showed reduced intensity of radiostaining. Skin prick tests and PK tests confirmed the reduced allergenicity of the heated fractions. These findings confirm the relative thermostability of the allergens in birch pollen. However, heating infers irreversible changes on the tertiary structure of the molecules, and these structural changes can be correlated to a quantitative reduction of allergenic reactivity.

Animals↗

PubMed1980

Analysis of commercial pollen extracts by enzyme determination. I. Comparison of rast-inhibition assay and enzyme titration for orchard grass, rye grass and short ragweed pollen extracts.

Allergenic potency assessed by the 50% RAST-inhibition endpoint titration was compared with enzyme titration for the analysis of commercial pollen extracts. Six different extracts of short ragweed, orchard grass and perennial rye grass were examined. A good correlation (p < 0.01) was found between the acid phosphatase titers and allergenic potency of short ragweed and perennial rye grass extracts. A similar correlation (p < 0.01) was observed between leucin amino peptidase titers and allergenic potency of short ragweed extracts. No correlation could be found with orchard grass extracts. These findings suggest that enzymatic activity may provide a method for the control of certain commercial pollen extracts.

Acid Phosphatase↗

PubMed1985

Pollen sporoplasts: dissolution of pollen walls.

4-Methylmorpholine N-oxide monohydrate (MMNO.H(2)O), a potent solvent for polysaccharides, is an effective vehicle for release of membrane-enclosed male gametophytes (sporoplasts) from spore walls. This release occurs in minutes when pollen (Lilium longiflorum Thunb.) is suspended in a melt of MMNO.H(2)O at 75 degrees C. Continued heating at 75 degrees C leads to distintegration of the exine ;shell' which coalesces into immiscible globules in the MMNO melt. These observations provide a general procedure for preparation of pollen sporoplasts and sporoplast outer membranes, and offer a new method for dissolving the sporopollenin component of the spore wall.

Journal Article↗

PubMed1983

Inhibition of the passive cutaneous anaphylaxis reaction to Dactylis glomerata pollen allergens by a purified component of this pollen.

By isoelectric focusing a protein fraction of pI 4.6 was isolated from a crude water-soluble extract of Dactylis glomerata pollen (SE). This fraction was neither immunogenic nor allergenic in BALB/c mice. In one week, this protein inhibited the mouse IgE-specific antibodies to the soluble extract as measured by PCA in rats and was therefore called Dactylis inhibitory protein (DIP). Two experimental approaches which lowered IgE anti-SE titer were undertaken. Pretreatment with DIP as well as injection of DIP after the last sensitizing injection with SE resulted in an inhibition of the circulating IgE antibody level to SE. For both experiments the regulation of the immune response touched only the IgE class, whereas the titers of anti-SE IgG, IgM, IgA antibodies were not modified. DIP treatment did not alter the IgE titers, measured by PCA, in the immune response to ovalbumin.

Allergens↗