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PubMed1992

Allergens from rye pollen (Secale cereale). I. Study of protein release by rye pollen during a 19-hour extraction process. Allergen identification.

We have studied the proteins and allergens released by rye pollen in the course of a 19-h pollen incubation process. Nearly 40% of the total extracted proteins were collected during the first 5 min, and most of them had a molecular weight less than 28 kDa. Between 5 and 30 min, 15% of the proteins from total extract were released, showing in the SDS-PAGE analysis an increase in which components moved close to 30 kDa standard. From 30 min to 19 h several extracts were collected. Electrophoretical profile of components from these extracts reveals that bands moving below 28 kDa were practically absent and those of 28 and 23 kDa became very intense. At the end of the process there was a rise of 67 kDa proteins. Dot-immunobinding and immunoblotting techniques reveal that allergens leave the rye pollen, for the most part, after 5 min incubation and are proteins with 28 kDa, 33 kDa, 48 kDa and 67 kDa molecular weights.

Allergens↗
PubMed1998

Double-blind, placebo-controlled evaluation of sublingual immunotherapy with standardized olive pollen extract in pediatric patients with allergic rhinoconjunctivitis and mild asthma due to olive pollen sensitization.

For evaluation of the efficacy and the safety of specific sublingual immunotherapy with high allergen dose, 66 children with seasonal asthma, rhinitis, and conjunctivitis due to sensitization to olive pollen were enrolled in a double-blind, randomized, placebo-controlled study between October 1994 and October 1996 in Greece. Thirty-four patients were randomly allocated to the active group, and 32 received placebo. Immunotherapy consisted of olive-allergen extracts (Stallergènes SA) administered sublingually pre- and coseasonally from January to July for 2 consecutive years. Serial concentrations from 1 to 300 IR. were used up to the maintenance dose of 20 drops of 300 IR daily. The cumulative dose for each patient was 300 times higher than in parenteral immunotherapy, and the cumulative dose of the major allergen Ole e 1 was 8.1 mg/2 years. The patients were assessed by clinical parameters (symptom and medication scores from patients' daily diaries) and immunologic measurements (specific IgE, IgG4, eosinophil cationic protein [ECP]) were performed. The actively treated patients had a significantly lower score for dyspnea (P<0.04 during the first season; P<0.03 during the second season). At the pollinic peak during the second year, a lower score of conjunctivitis was recorded (P<0.05) in the actively treated patients. The analysis of intragroup evolution showed that the total score of rhinitis increased significantly during the pollinic peak in the group under placebo, whereas there was no symptomatic peak for the same period in the group under active treatment. However, the difference between the groups was not significant. The medication score did not differ significantly between the groups. Oral steroids were the only variables with a P value near the significance level (P=0.06) in favor of the actively treated group. A significant decrease in skin reactivity was recorded in the active group after 2 years of treatment. No significant variation in specific IgE and IgG4 was detected. A significantly lower level of serum ECP was observed at the pollinic peak in the actively treated patients during the first pollen season (P=0.01), but this was not confirmed the second year when the ECP levels doubled in both groups without correlation to the clinical findings. Tolerance was excellent with only a few minor side-effects reported. In conclusion, high-dose specific sublingual immunotherapy appears to be safe and effective in improving mild seasonal asthma and conjunctivitis linked to olive-pollen sensitization.

Administration, Sublingual↗
PubMed2004

Evaluation of recombinant and native timothy pollen (rPhl p 1, 2, 5, 6, 7, 11, 12 and nPhl p 4)- specific IgG4 antibodies induced by subcutaneous immunotherapy with timothy pollen extract in allergic patients.

BACKGROUND: Allergen immunotherapy is a widely accepted treatment for IgE-mediated allergies. The evaluation of immunotherapy-induced IgG4 antibodies based on allergen extract is questionable because the amount of allergen-extract-specific IgG4 to individual disease-eliciting allergens cannot be determined using crude allergen extracts. In this study, we examined the specific IgE and IgG4 serum binding profiles to individual Phleum pratense allergens in grass-pollen-sensitive patients who had received grass-pollen-specific immunotherapy (SIT). METHODS: The study included 33 patients from North-West Italy. All suffered from seasonal rhinoconjunctivitis and/or asthma. A modified "cluster" regimen of injections of a standardized aluminium-adsorbed P.pratense extract, with once-weekly visits and 10 injections for 5 weeks followed by 3 weeks of maintenance injections was instituted. Patients' sera were analyzed for specific IgE and IgG4 reactivity to individual P. pratense allergens (recombinant Phl p 1, Phl p 2, Phl p 5, Phl p 6, Phl p 7, Phl p 11, Phl p12 and native Phl p 4) and natural P. pratense extract using the Pharmacia CAP system. RESULTS: IgE reactivities to new allergen components were not detected by CAP in treated patients after 15 weeks and a cumulative dose of approximately 65 microg of the major allergen Phl p 5. Patients lacking specific IgE reactivity towards individual allergens at the start of SIT did not produce significant levels of specific serum IgG4 to serum IgE-negative allergens. On the other hand, an increase in specific IgG4 only to allergens to which patients were previously sensitized was observed. Significant increases in specific IgG4 levels to rPhl p 1 (p < 0.05), 2 (p < (0.01), 5 (p < 0.0001), 6 (p < 0.0001), 7 (p < 0.05), 11 (p < 0.05) and nPhl p 4 (p < 0.01) were observed after P. pratense extract immunotherapy. No significant rPhl p 12-specific IgG4 antibody increase was documented after treatment. CONCLUSION: These findings suggest that Phl p 12 was underrepresented in the extract used, as indicated by the low specific IgG4 response induced by this grass-pollen-specific vaccine. Thus, the simple detection of specific serum IgG4 antibodies a few weeks after the start of SIT could represent a valuable tool to estimate the presence of relevant allergens in a given immunotherapeutic allergen extract.

Adolescent↗
PubMed2007

Immunotherapy with an aluminum hydroxide-adsorbed Juniperus ashei foreign pollen extract in seasonal indigenous cypress pollen rhinoconjunctivitis. A double-blind, placebo-controlled study.

BACKGROUND: The efficacy of standardized Juniperus ashei extract was assessed in patients with allergic rhinoconjunctivitis due to European cypress pollens. METHODS: Forty adults with European cypress-allergic rhinoconjunctivitis were randomized to receive immunotherapy or a matched placebo. Specific immunotherapy was performed with a standardized, aluminum hydroxide-adsorbed J. ashei extract with a potency of 100 IR (arbitrary index of reactivity) containing 54 microg of Jun a 1/ml (Alustal, Stallergenes, France). Subcutaneous injections started in October 2000. The maintenance dose was 0.30 ml of the 100-IR concentration per month. Rhinitis and conjunctivitis symptoms were rated according to a 4-point score. RESULTS: Seventeen patients from the treated group and 15 patients from the placebo group completed year 2001; 14 in each group completed year 2002. A statistically significant improvement (41%, p < 0.02) in the conjunctivitis symptom score was observed in actively treated patients compared to the placebo group at the peak of the 2001 pollen season. Improvement in rhinitis (17%) was not significant. This significant improvement was greater at the peak of the 2002 pollen season (63%, p < 0.01). CONCLUSIONS: This study therefore indirectly validates the concept of treatment by major allergen because J. ashei is absent from the region in which this study was conducted.

Adsorption↗
PubMed1990

Grass pollen allergens: detection on pollen grain surface using membrane print technique.

A microtechnique is described to visualize pollen allergens by making prints from the surface of mature grains on nitrocellulose membrane. The allergen Lol p I is detected by specific monoclonal antibodies or specific IgE from sera of grass-pollen-allergic patients, visualized by the immunoperoxidase method. This technique offers the potential to study the distribution of individual allergens in aerobiology samples by bright-field microscopy, and to determine the sensitivity spectra of allergic individuals to known and unknown pollen sources.

Allergens↗
PubMed2000

Exposure to Japanese cedar pollen in early life and subsequent sensitization to Japanese cedar pollen.

The effect of exposure to Japanese cedar pollen (JCP) in early life on subsequent sensitization to it was evaluated. Specific IgE antibody to JCP was examined in 440-504 school children in a rural town each year during 1995-98. The amount of dispersed pollen measured by a Durham sampler widely ranged from 165 to 5941 grains/cm2/year during this period. The amount had been measured during the period of 1982-91 in which these children were born, and it also widely ranged from 148 to 8566 grains/cm2/year. Children born during November to January, who were exposed to JCP within 6 months of age, increased at the risk of sensitization to JCP, especially severe sensitization, relative to those born in the other months. Age-adjusted prevalence rate ratio (RR) of having a JCP-IgE > or = 15 U/ml (control; < 0.35 U/ml) for children born in December to February relative to children born in the other months was 1.74 (95% confidence interval; 1.06-2.87, examined in 1998), and for those born in November to January was 1.57 (95% CI; 1.00-2.46, examined in 1997). The risk of sensitization to JCP was low for those born in May to July (RR = 0.42, 95% CI; 0.19-0.93, examined in 1998). There was also a strong correlation between the amount of the dispersed pollen during the period of 2-6 months after birth and the prevalence of sensitization to JCP.

Adolescent↗
PubMed1999

[Prevalence rate of allergy to Japanese cedar pollen in Tokyo--from field investigation in 1996 by Tokyo Japanese Cedar Pollen Allergy Measurements and Review Committee].

Prevalence rate of allergy to Japanese cedar pollen was investigated by means of questionnaires and immunological examinations in residents of 3 regions in Tokyo (Akiruno city, Chofu city, and Ota ward). The results were as follows: 1) the prevalence rate of allergy to Japanese cedar pollen in Tokyo was 25.7% in Akiruno city, 21.1% in Chofu city, and 17.7% in Ota ward in 1996; 2) stratification by the age reveal that the prevalence rate was the highest in the age group of 30 to 44, and the group showed prevalence rate of above 40% in Akiruno city; 3) overall prevalence rate of Tokyo was 19.4% in 1996, which was approximately twice as much as that in 1985; 4) increase in prevalence rate in the recent decade was most significant in Akiruno city where the largest number of Japanese cedar pollen was detected in the air, and prevalence rate tended to rise in the young group of 14 or less.

Adolescent↗
PubMed2002

[Gramineae pollen dispersal and pollinosis in the city of Hisai in Mie Prefecture. A 14-year study of gramineae pollen dispersal and cases of sensitization to gramineae experienced at an allergy clinic over a 15-year period].

(1) A study of Gramineae pollen dispersal was conducted in Hisai, Mie Prefecture from 1987 to 2000; Gramineae pollen counts did not increase, with an average of 133 during the 14 years. There was two or three peaks in the annual dispersal period, suggesting that symptoms in late May, late July to early August, and late September are important for diagnosis of Gramineae pollinosis in this region. (2) Changes in the frequencies of sensitization to Gramineae pollen and Gramineae pollinosis were studied in 756 allergy clinic patients with nasal allergies examined over a 15-year period from 1986 to 2000. Results for the frequency of sensitization every 5 years came to 7.9, 22.4, and 23.3% and that of Gramineae pollinosis were 4.8, 10.3, and 12.1%. Both frequencies tended to increase after 1990. Out of 135 patients with sensitization to Gramineae, 88.1% had multiple sensitization to Japanese cedar and 54.8% had sensitization to Japanese cedar as well as house dust or mites. Patients sensitized only to Gramineae accounted for 5.9% of total patients. About half of the patients with sensitization to Gramineae developed pollinosis. As the CAP RAST score increased, so did the increase of pollinosis.

Adolescent↗
PubMedDate not supplied

[Allergological study of pollen of mango (Magnifera indica) and cross reactivity with pollen of piru (Schinus molle)].

With immediate skin test, we studied hypersensitivity to pollen of mango (Magnifera indica), and its probability cross-sensitivity to pollen of pirú (Schinus molle), from anacardaceae class. We have made one extract of Magnifera indica, with 0.485 mg per 100, of protein nitrogen. With this extract we carry out skin test on non-atopic subjects and atopic subjects with bronchial asthma and or allergy rhinitis and common stock of aerealergen. (Freeman stock). We studied seventy-one subjects, both sexes, from 14 to 40 years old. The prevalence of cutaneous sensitive to mango (Magnifera indica) were 66%, and sensitivity from Schinus molle were 31%. Those with sensitivity Schinus molle, were sensitivity from Magnifera, too. Statistical analyisis of correlation, with McNemar Test was p 0.001 under the conditions of the study we conclude that there are hypersensitivity to pollen of Magnifera, and there are cross-reactivity with Schinus molle. Therefore mangifera should be included within the stock of skin test in the areas were this anacardaceae is present.

Adolescent↗
PubMed1987

[Allergologic-immunochemical study of tree and bush pollen. III--Cross reactions of human IgE antibodies with various tree pollen allergens].

Partial identity between the major allergens of birch, beech, alder, hazel and oak pollen extract could be identified by means of RAST-, ELISA- and CRIE-inhibition as well as further types of crossed immunoelectrophoresis. It seems that spring time pollinosis in our region of Central Europe is caused principally by the major allergen of birch pollen. Cross reactivity between grass and tree pollen could not be found. Patients with symptoms in spring and summer time are double sensitized.

Antibody Specificity↗
PubMedDate not supplied

Analysis of commercial pollen extracts by enzyme determination. II. Determination of the contamination of pollen extracts with plant matter.

The contamination of pollen extracts by plant matter was studied by means of enzymes. Beta-glucosidase was detected only in flowers and leaves by using the Api-Zym System. 29 pollen extracts were prepared after microscopic control. Among these 23 did not contain any plant matter and no beta-glucosidase activity could be detected. In contrast, 6 extracts were contaminated with plant matter and in every case beta-glucosidase activity was found 34 commercial pollen extracts completed the study. beta-glucosidase activity was present in 8 of them. It may be postulated that glucosidase activity detected using 2- naphthyl - beta D-glucopyranoside as a substrate, can be correlated with the presence of plant matter from the raw material from which the extract was prepared. This contamination may be important to determine since some additional sensitization may occur with such extracts during immunotherapy.

Drug Contamination↗
PubMed1981

Comparative studies on tree-pollen allergens. I. Isolation and partial characterization of a major allergen from birch pollen (Betula verrucosa).

A major allergen from birch pollen (Betula verrucosa) was isolated by a combination of gel permeation chromatography and preparative isoelectric focusing, and was found adequate for further immunological and chemical characterization. The crude aqueous pollen extract was eluted in 6 UV-absorbance peaks from an Ultrogel AcA-54 column. The main allergenic components were detected in two peaks of molecular weights 29,000 (BV4) and 40,000 daltons, respectively. BV4 contained one quantitatively dominating antigen as assessed by crossed immunoelectrophoresis, and was selected for further purification by isoelectric focusing. Column isoelectric focusing of BV4 gave four protein fractions with pI values corresponding to 5.18, 5.42, 5.76, and 5.94. The pI 5.18 protein was quantitatively dominating. This protein inhibited the radio-allergosorbent test in 8 out of 10 sera from birch pollen-sensitive individuals. Using a serum pool, the inhibition curve of BV4 pI 5.18 was parallel to that of the crude extract and gave a similar maximum inhibition. This allergen was shown to be homogeneous as judged by a single, symmetrical precipitate in crossed immunoelectrophoresis, one band in high-volt electrophoresis and a sharp strip in thin-layer analytical electrofocusing. Amino acid analysis showed a high content of serine. The N-terminal analysis as well as the carbohydrate concentration were preliminarily reported and will be confirmed in future studies.

Allergens↗
PubMed1987

An enzyme-linked immunosorbent assay (ELISA) for in vitro pollen growth based on binding of a monoclonal antibody to the pollen tube surface.

An indirect method of quantifying in vitro pollen growth has been developed. This is based on the finding that a monoclonal antibody (PCBC3), which has a primary specificity for alpha-l-arabinofuranosyl residues, binds to the surface of in vitro grown pollen tubes of the ornamental tobacco, Nicotiana alata. This binding was quantified using an enzyme linked immunosorbent assay (ELISA). The method was used to determine the effects of 2-deoxy-d-glucose and nonanoic acid on in vitro pollen growth.

Journal Article↗
PubMed2004

Vaccines for birch pollen allergy based on genetically engineered hypoallergenic derivatives of the major birch pollen allergen, Bet v 1.

BACKGROUND: We have recently engineered recombinant derivatives of the major birch pollen allergen Bet v 1 (rBet v 1 fragments and trimer) with strongly reduced allergenic activity. OBJECTIVE: The aim of this study was the in vivo characterization of potential allergy vaccines based on Al(OH)3-adsorbed genetically modified rBet v 1 derivatives in mice. METHODS: BALB/c mice were immunized either with courses of nine injections of increasing doses of Al(OH)3-adsorbed rBet v 1 wild-type, rBet v 1 fragments, rBet v 1 trimer or Al(OH)3 alone in weekly intervals or with three high-dose injections applied in intervals of 3 weeks. Humoral immune responses to rBet v 1 wild-type and homologous plant allergens were measured by ELISA and Western blotting, and the ability of mouse antibodies to inhibit the binding of allergic patients IgE to Bet v 1 was studied by ELISA competition experiments. RESULTS: In both schemes, hypoallergenic rBet v 1 derivatives induced low IgE but high IgG1 responses against rBet v 1 wild-type. The IgG1 antibodies induced by genetically modified rBet v 1 derivatives cross-reacted with natural Bet v 1 and its homologues from alder (Aln g 1) as well as hazel (Cor a 1) and strongly inhibited the binding of birch pollen allergic patients' IgE to Bet v 1 wild-type. CONCLUSION: Genetically modified hypoallergenic rBet v 1 derivatives induce blocking antibodies in vivo. Their safety and efficacy for the treatment of birch pollen and associated plant allergies can now be evaluated in clinical immunotherapy studies.

Allergens↗
PubMed1976

The cross-reactivity of IgE antibodies with pollen allergens. I. Analyses of various species of grass pollens.

Atopic patients with histories of grass pollen allergy often are sensitive to a variety of species of grasses. Using a serum pool from patients sensitive to June grass, we analyzed the reactivity of IgE antibodies to seven grasses by the radioallergosorbent test. Extracts were analyzed for their inhibitory activities with solid-phase allergens prepared from all of the grass pollen. Also samples of serum were exhaustively absorved with solid-phase allergens and the supernatants tested to determine the reactivity of the remaining IgE antibodies. Three patterns of reactivity were observed: (1) June, orchard, meadow fescue, and perennial rye grasses displayed similar reactivity in both inhibition and absorption studies; (2) sweet vernal and Bermuda grasses were considerably less reactive with the serum pool, indicating that they lacked antigenic determinants possessed by the other grasses; and (3) timothy grass possessed unique antigenic determinants. Knowledge of these patterns of cross-allergenicity is of importance for diagnosis and treatment of sensitive patients as well as for in vitro standardization of extracts.

Absorption↗
PubMed1988

Immunoblotting of pollen proteins: Tween 20 mediates nonspecific binding of gammaglobulins to immunoblots of oak-pollen proteins.

Oak-pollen extracts, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred onto nitrocellulose, demonstrated extensive banding in the absence of human sera or with nonatopic sera with both a horseradish peroxidase-conjugated goat antihuman IgE and 125I-labeled rabbit antihuman IgE. When the nonionic detergent Tween 20 was removed from all incubations and washes, negative banding appeared, corresponding to bands that were clearly visible when Tween 20 had been used as a blocking agent. When Tween 20 was reintroduced, the bands reappeared. These results demonstrate that, although Tween 20 is effective in blocking unbound sites on nitrocellulose membranes, this and other nonionic detergents may cause nonspecific binding to immunoblots of oak-pollen proteins. Thus, care must be used in the interpretation of immunoblots to detect allergens when Tween 20 is used as the blocking agent.

Animals↗
PubMed1997

Recombinant Bet v 1, the major birch pollen allergen, induces hypersensitivity reactions equal to those induced by natural Bet v 1 in the airways of patients allergic to tree pollen.

BACKGROUND: Atopic allergens produced by recombinant DNA methods are promising tools for diagnosis and therapy of Type I allergy. To evaluate the immunologic properties of these molecules, it is necessary to compare them with natural allergens in vitro and in vivo. OBJECTIVE: The study was carried out to determine whether the potency of recombinant Bet v 1 (rBet v 1) is comparable to that of natural Bet v 1 (nBet v 1) in inducing allergic reactions in the nose and bronchi. METHODS: Thirteen patients allergic to birch pollen with bronchial asthma and/or rhinitis were investigated. Skin prick tests and nasal and bronchial challenges were performed with rBet v 1 and nBet v 1. RESULTS: In patients allergic to birch pollen, both allergens induced comparable skin reactions. In subjects with rhinitis rBet v 1 was equally potent in inducing nasal reactions (mean PD(+60)NR +/- SD, 10.48 +/- 17.42 microg vs 7.98 +/- 8.9 microg, p > 0.05). In patients with asthma, rBet v 1 was equally potent in inducing bronchial reactions (PD20 FEV1, 0.81 +/- 1.74 microg vs 0.62 +/- 1.44 microg, p > 0.05) as nBet v 1. CONCLUSION: No significant differences were observed between natural and recombinant allergen. We conclude that allergens produced by recombinant techniques can induce typical allergic reactions in important target organs of Type I allergy: the nose and bronchi.

Adult↗
PubMed1998

Identification of Cannabis pollens using an allergic patient's immunoglobulin E and purification and characterization of allergens in Cannabis pollens.

Cannabis pollen allergens were detected using the serum of an allergic patient. The allergens were then purified by sequential column chromatography (including DE52 cellulose and phenyl-Sepharose CL-4B) and preparative HPLC. The molecular weight of the allergens were determined as 10,050 and 13,706 by matrix-assisted laser desorption/ionization time of flight mass spectrometry. We utilised Western blotting and development of an enzyme-linked immunosorbent assay for the detection of Cannabis pollen allergens.

Allergens↗