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PubMed2004

Clinical efficacy of microencapsulated timothy grass pollen extract in grass-allergic individuals.

BACKGROUND: Conventional allergen immunotherapy is clinically effective in reducing the symptoms of allergic rhinitis and asthma. It differs from other pharmacotherapies in that it can induce long-term clinical remission of these diseases. However, it requires years of treatment and is associated with serious allergic reactions. OBJECTIVE: To evaluate the safety, clinical efficacy, and immunologic mechanisms of immunotherapy with an oral, microencapsulated form of timothy grass allergen. METHODS: In this double-blind, placebo-controlled study, 24 patients aged 19 to 55 years with grass pollen allergy were randomized to receive either microencapsulated timothy grass pollen extract or placebo once a day for 10 weeks. The dose of study drug was doubled weekly. Safety was evaluated through weekly visits, daily symptom diaries, and routine laboratory tests. Efficacy was evaluated by comparing medication use and symptoms scores during peak grass pollen season before and after treatment. Allergen-specific T-cell responses, cytokine production, and IgG, IgE, and skin reactivity were measured to evaluate immunologic mechanisms. RESULTS: Eleven of 12 patients in the active treatment group had a decrease in the combined medication and symptom score, but only 4 of 10 patients in the placebo group had a decrease in scores. The proliferative response to timothy grass was reduced by at least 30% in 9 of the 12 grass-treated patients, but only 3 of 11 placebo patients had a proliferative response reduction. Timothy grass-induced interleukin-5 messenger RNA was reduced in the active group, but not in the placebo group. There were no significant changes in either group in IgG, IgE, and skin reactivity. CONCLUSIONS: Oral immunotherapy with microencapsulated allergen induces a form of immunologic tolerance to the allergen and is a safe, efficient, and effective method of allergen immunotherapy.

Administration, Oral↗
PubMed1989

Transfer of Cs-137 from grass and wilted grass silage to milk of dairy cows.

Deposition of radiocaesium from the Chernobyl reactor accident on the Netherlands made it possible to collect contaminated fresh grass and first cut wilted grass silage. These contaminated roughages were used in transfer experiments with lactating dairy cows to determine transfer coefficients and half-lives for Cs-137 in milk. The experimental design was based on three consecutive periods: a preliminary period to determine the background concentration of the isotope in milk, a contamination period to determine the magnitude of accumulation and finally a depletion period to measure the rate at which the activity concentration of Cs-137 in milk declined after continuous feeding. The average transfer coefficient (Fmilk) for cows fed on contaminated dried grass under steady-state conditions was 0.002 d/kg and for cows fed on slightly contaminated second cut fresh grass 0.006 d/kg. The highest transfer coefficients were obtained for cows fed on contaminated grass silage for 119 days, which also included the dry period of about two months. For the first five days after calving the Fmilk values varied from 0.0066 to 0.0091 d/kg. There were no significant differences in transfer coefficients between cows in early lactation (third month of lactation), cows in late lactation (the last month of the lactation period) and cows fed on both contaminated grass silage and uncontaminated maize silage simultaneously. Half-life values for the rate of decline of the isotope in milk during the depletion period were estimated on the basis of a mathematical model with two exponential components. These components were characterized by half-lives of 0.5 to 3.5 days and 10 to 46 days.

Accidents↗
PubMed1995

T-cell epitopes of Phl p 1, major pollen allergen of timothy grass (Phleum pratense): evidence for crossreacting and non-crossreacting T-cell epitopes within grass group I allergens.

BACKGROUND: The use of peptides representing T-cell epitopes of allergens is a modern concept for improvement of specific immunotherapy. A prerequisite for this approach is the identification of T-cell epitopes of atopic allergens. METHODS: T-cell lines and 40 T-cell clones (TCC) specific for Phl p 1, the group I allergen of timothy grass (Phleum pratense), were established from the peripheral blood of nine patients allergic to grass pollen and mapped for epitope specificity by using overlapping dodecapeptides. Phenotype and cytokine production profile of TCC were investigated. Representative TCC were analyzed for HLA-restriction, T-cell receptor V beta gene usage, and crossreactivity with grass pollen extracts from Dactylis glomerata, Poa pratensis, Lolium perenne, Secale cereale, and selected amino acid sequence-derived peptides. RESULTS: Patients displayed IgE binding to all grass species investigated. Forty TCC were established. Fifteen T-cell epitopes could be identified on Phl p 1. Of 40 TCC, 39 displayed the helper cell (Th) phenotype; one clone was CD8+. Specific stimulation induced a Th2-like type of cytokine production in 20 of 39 TCC. Crossreactivity studies revealed crossreacting and non-crossreacting T-cell epitopes. CONCLUSION: Phl p 1, a major grass pollen allergen, harbors multiple T-cell epitopes. Species-specific and crossreacting T-cell epitopes exist among group I allergens of grasses. Epitope recognition patterns could not be correlated with particular HLA haplotypes. A restricted T-cell receptor V beta gene usage was not observed.

Allergens↗
PubMed1993

Comparison of four grass pollen species concerning their allergens of grass group V by 2D immunoblotting and microsequencing.

The identification and characterization of allergenic components is a vital step towards improving diagnosis and therapy. Members of the grass family (Poaceae) reveal a high cross-reactivity among each other caused by the close phylogenetical relationship. In order to investigate the variability between allergenic components, we studied the allergen grass group V, one of the major allergens. Pollen extracts of 4 different tribes (timothy grass (Phleum pratense)--Agrostidae, perennial rye grass (Lolium perenne)--Festuceae, meadow velvet (Holcus lanatus)--Aveneae, and rye (Secale cereale)--Triticeae) of the Festucoideae subfamily were separated by 2D PAGE and investigated by immunoblotting using patients' poolserum and monoclonal antibodies (raised against group V allergens of timothy grass pollen). The antibodies identify different allergens in the four grass species. The components vary from 30-50 kDa and pI 4.8-7.0. The eight NH2-terminal amino acids were determined and indicated high similarities between the different components. These results cast doubt on the suitability of classifying allergens into groups based only on their molecular mass, isoelectric point and N-terminal sequence analysis. It suggests to classify allergens according to their IgE-reactive epitopes.

Allergens↗
PubMed2001

Milk production of fall-calving dairy cows during summer grazing of grass or grass-clover pasture.

Milk production of fall-calving dairy cows during subsequent summer grazing was evaluated in two consecutive years using a total of 80 mid- to late-lactation Holsteins. Cows calved during September and October and grazed from April to August in the following year. In yr 1, 27 cows grazed a native grass pasture and 13 cows grazed a native grass-clover mixed pasture containing 26% red clover and white clover. In yr 2, 40 cows grazed native grass pasture as one group. Also, cows in yr 2 were administered bovine somatotropin, whereas in yr 1, no bST was used. Grazing cows also were fed concentrate supplements at 6.2 kg/d of dry matter (DM) in yr 1 and 7.9 kg/d of DM in yr 2 to provide 35 to 40% of total intake. Average daily milk during the grazing period decreased 3.6 kg in yr 1 and 7.7 kg in yr 2 when compared with milk yield extrapolated from the lactation curve established 10 wk before being turned out to pasture. Estimated DM intake during grazing was also less than what would have been expected had cows continued on a total mixed ration in confinement. Cows grazing the mixed pasture of grass and clover yielded 1.3 kg/d more milk than those grazing the grass pasture in yr 1. A decrease in milk resulting from the change from total mixed ration fed in confinement to grazing supplemented with concentrates was not avoided with these mid- to late-lactation cows, but the cumulative loss over the lactation was less than with early lactation cows in a companion study. Clover enhances the grazing value of pasture when grown with grasses.

Animal Feed↗
PubMedDate not supplied

Manure management effects on grass production, nutritive content, and soil nitrogen for a grass silage-based dairy farm.

Legislation in the United States has recently focused on improving water quality by establishing management practices that limit the quantities of nutrients entering the water supply. Timely application and quantification of the amount of manure applied throughout the grass-growing season can reduce the loss of nutrients into ground or surface water while improving the quality and quantity of grass harvested. During the 2001 and 2002 growing seasons, we measured the effects of different manure application rates on grass yields, grass nutritive value, and soil chemistry on a dairy farm. On-farm estimates of manure N were combined with yield estimates and forage quality measures to evaluate the effects of varying levels of manure application. Yield estimates, N content of grass, and the amount of N in soil and manure were monitored at each cutting for plots amended at different manure application rates. There are three major outcomes of this evaluation: (i) new grass seedings were at higher risk of elevated levels of nitrate N in forage; (ii) increased forage nitrate N at harvest was associated with malfermented silage and increased levels of ammonia N, which resulted in less efficient use of metabolizable protein for milk production; and (iii) increased understanding of N cycling between manure, soil, and plant provided an opportunity to reduce purchased fertilizer.

Animals↗
PubMed1986

A comparison of the immune response to immunotherapy with polymerized grass allergen and monomeric grass allergen.

This study compares the immune response of ten patients treated with polymerized grass (PG) immunotherapy with that of 12 patients treated with conventional monomer grass (MG) immunotherapy. The patients treated with PG immunotherapy received a mean cumulative dose of 44,840 protein nitrogen units (PNU), and those treated with MG immunotherapy received a mean cumulative dose of 46,083 PNU. Total antibody binding of perennial rye grass groups I, II, and III (RGGI, RGGII, and RGGIII) was measured in the serum of each individual. In addition, IgG titers to partially purified extracts of Bermuda, timothy, and orchard grass were also determined. A significant increase in the total antibody binding of RGGI, as well as an increase in the mean IgG titer to Bermuda, timothy, and orchard grass was demonstrated after treatment in the patients who had received PG immunotherapy. The mean total antibody binding of RGGII and RGGIII also increased in these patients, although not significantly. No significant difference in the mean total antibody binding of RGGI, RGGII, or RGGIII nor in the mean IgG titer to the three grass extracts was found in the sera of the two groups of patients. This study demonstrates a comparable immune response between immunotherapy with PG and MG, and retention of antigenic determinants during the polymerization process.

Allergens↗
PubMed1994

A Common Allergenic Epitope of Bermuda Grass Pollen Shared by Other Grass Pollens.

The present study disclosed the cross-reactivity between Bermuda grass pollen (BGP) and other grass pollens using monoclonal antibodies (MAbs) and polyclonal antiserum. MAb 9-13, directed against a group of minor allergens of BGP (Cyn d Bd68K, 48K, 38K) was found to cross-react with extracts of ten other grass pollens. Immunoblotting assays illustrated that MAb 9-13 cross-reacted with multiple components of most of these pollens, and the major cross-reactive components had molecular weights of 29-36 kD. The crossreactivity between BGP and Lol pI, the group I allergen of rye grass pollen, was further evaluated; Lol pI was recognized by MAb 9-13, but not by our MAbs/polyclonal antiserum against Cyn dI, the major allergen of BGP. These results suggest that the epitope recognized by MAb 9-13 is a common (C) epitope shared by Lol pI and Cyn d Bd68K, 48K, 38K, and Cyn dI does not share significant antigenicity with Lol pI. In a modified radio-allergosorbent test, IgE antibodies in the serum of BGP-allergic patients reacted mildly with C-epitope-bearing components of both BGP and rye grass pollens, and this binding could be blocked specifically by MAb 9-13. This suggests that in addition to an antigenic cross-reaction, the C epitope can also lead to an allergenic cross-reaction. Copyright 1994 S. Karger AG, Basel

Journal Article↗
PubMed1986

A double-blind placebo-controlled trial of polymerized whole grass administered in an accelerated dosage schedule for immunotherapy of grass pollinosis.

Forty-four patients were entered into a study of the efficacy and safety of individually polymerized grass (IPG) immunotherapy with an accelerated dosage schedule. Patients were paired on the basis of cutaneous end point titrations to timothy, orchard, and Bermuda grass-pollen extracts. In a double-blind manner, one patient in each pair was treated in nine weekly visits with 13 injections that totaled 24,000 PNU of each grass to which the patient had cutaneous reactivity. The other patient in each pair received caramelized glucose histamine placebo. Symptom and medication score sheets were completed by 33 patients each day of the grass season. Blocking antibody rose significantly in the IPG-treated group but was unchanged in the placebo-treated group. By Wilcoxon paired signed-rank test, the symptom medication scores in the IPG-treated group were significantly lower than those in the placebo-treated group. There were no systemic reactions and no clinically significant changes in routine laboratory tests in either group. In summation, this study demonstrates the safety, immunogenicity, and efficacy of IPG therapy in an accelerated dosage schedule for treatment of grass pollinosis.

Adult↗
PubMed1989

Fluctuation of the number of CD-1(T6)-positive dendritic cells, presumably Langerhans cells, in the nasal mucosa of patients with an isolated grass-pollen allergy before, during, and after the grass-pollen season.

A monoclonal antibody against CD-1(T6) was used for studies in the nasal mucosa of patients with isolated grass-pollen allergy to determine whether the number of CD-1-positive cells, presumably Langerhans cells, depends on the season in which the nasal biopsy is performed. An earlier study had demonstrated that during the grass-pollen season, there are significantly more CD-1-positive cells in nasal mucosa of patients with isolated grass-pollen allergy than in nonallergic control subjects without nasal complaints. During the grass-pollen season, the nasal epithelium of patients with an isolated grass-pollen allergy demonstrated significantly more CD-1-positive cells than before and after the season. Before and after the season, the number of CD-1-positive cells in epithelium of the allergic patients was not significantly greater than the corresponding number in epithelium of nonallergic subjects without nasal complaints.

Adolescent↗
PubMed1984

Grass pollen immunotherapy: a single year double-blind, placebo-controlled study in patients with grass pollen-induced asthma and rhinitis.

Fifteen grass pollen--sensitive asthmatic patients were selected from 200 patients with grass pollenosis on the basis of positive SPTs and RASTs that were restricted to grass pollens (except Bermuda grass), no previous IT, and residence and occupation in an area monitored by serial pollen counts. They underwent a double-blind trial of specific IT with a mixture of three grass pollen--aqueous extracts (velvet, sweet vernal, and timothy) or placebo. After 10 mo, the mean maintenance dose of pollen extract (assayed by RAST inhibition) in eight actively treated patients was 6000 RAST units (range 3000 to 8000) and the mean total dose was 18,700 RAST units (range 10,200 to 30,000). Results were assessment done by the following clinical and immunological data: (1) during the pollen season, daily symptom scores; (2) PD 20% FEV1, IgE antibody to timothy by RAST in serum and in nasal secretions, serum IgG antibody to purified timothy allergen D by solid-phase radioimmunoassay, and the four IgG subclass antibodies by enzyme immunoassay were all measured before treatment and before and after the pollen season. Symptom scores of both treated patients and controls correlated with pollen counts (R = 0.88, p less than 0.05 and R = 0.71, p less than 0.05, respectively). There was a significant difference between the mean symptom score values of treated patients versus controls (Kruskal-Wallis test, p less than 0.001). No significant differences or changes either in the PD 20% FEV1 or IgE antibody to timothy in serum and nasal secretions were found in the two groups before or after IT.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1979

Species specific grass pollen sensitivity: diagnosis and treatment with single grass species Allpyral vaccines.

Skin test titrations and nasal provocation tests in sixty patients with hay fever showed specific reactions to extracts of individual grass species. There was, however, no correlation between skin and nasal sensitivity. Repeat testing after treatment with Allpyral vaccines consisting of only the grass species to which the nasal reaction was most severe, or only one of several pollens to which reactions were equally severe, showed marked diminution of skin and nasal sensitivity not only to the single pollen used for immuno-therapy but to all five common pollens used in the Allpyral grass mix. Clinical results seemed much improved as compared with results in the same year for Allpyral five grass mix vaccines, especially in the case of patients treated with Timothy, rye, or cocksfoot. It was concluded that these three grassees were to be preferred for treatment in England, and that these grasses contain common allergens.

Dose-Response Relationship, Immunologic↗
PubMed1997

Immunological changes during specific immunotherapy of grass pollen allergy: reduced lymphoproliferative responses to allergen and shift from TH2 to TH1 in T-cell clones specific for Phl p 1, a major grass pollen allergen.

BACKGROUND AND OBJECTIVE: The mechanisms operative in specific immunotherapy (SIT) of Type I allergy are not completely understood. In the present study we evaluated immunological changes during SIT in pollinosis. METHOD: Eight patients suffering from pollinosis (monosensitized to grass pollen) were treated with conventional SIT. All subjects had IgE specific for Phl p 1, a major allergen of timothy grass. In vitro changes in the immunological reactivity to grass pollen extract and to recombinant Phl p 1 were evaluated. Subjects were examined at three occasions: before, after 3 months and after 1 year of SIT. RESULTS: Serological analysis revealed a marked increase of grass pollen- and Phl p 1-specific IgG, titres of specific IgE did not change significantly. Lymphoproliferative responses to grass pollen extract and rPhl p 1 were reduced already after 3 months of treatment. Accordingly, the cloning efficiency for Phl p 1-specific T-cell clones (TCC) dropped markedly in all patients. The majority of allergen-specific TCC raised before SIT revealed a TH2-like pattern of cytokine production, TCC established after SIT revealed TH1 characteristics. This shift was due to a decrease in IL-4 rather than an increase in IFN-production by T cells. Investigations of the epitopes recognized by T cells before and after SIT did not reveal the outgrowth of new ('protecting') specificities. We could not observe induction of allergen-specific CD8+ lymphocytes (supressor cells). CONCLUSION: Our data indicate that -- on the level of TH lymphocytes -- SIT induces tolerance to the allergen and a modulation of the cytokine pattern produced in response to allergen stimulation.

Adult↗
PubMed1984

Rationale for specific immunotherapy of grass pollen allergy with extracts of rye pollen. Skin test reactivity and immunochemical relationship between pollen allergens from rye and other common grasses.

In immunotherapy of grass pollen allergy, an extract of rye (Secale cereale) is often included. The aim of this study was to investigate by skin prick test (SPT) and immunochemical methods whether rye pollen contains specific allergens justifying the use of this extract separately. Twenty grass pollen allergic patients were skin prick tested with a dialysed freeze-dried raw extract of rye pollen (Sc), timothy extract (Soluprick SQ, 1 HEP) and two other rye extracts (Soluprick). Sera from the patients were RAST-tested using Sc and timothy (Pp). CRIE was performed using Sc and rabbit-anti grass (aNG) antibodies. The antigenic relations between rye and common grasses were investigated by CLIE using Sc and aNG as references, and by RAST inhibition. The ability of aNG to absorb the allergen activity of Sc was also tested. Significant correlations were found between timothy and rye when compared by means of SPT and RAST. The immunochemical analyses did not reveal any rye antigens containing rye epitopes only. However, the possibility of rye antigens with several epitopes, of which at least one is specific for rye, could not be excluded. Clinical symptoms supposedly elicited by rye alone can be explained quantitatively by the strongly time-limited and concentrated natural exposition. Diagnosis and treatment can, however, be performed with extracts of common grasses.

Adolescent↗
PubMed1985

Anti-Bermuda grass RAST binding is minimally inhibited by pollen extracts from ten other grasses.

This article addresses the question, Does sensitization with Bermuda grass pollen generate IgE antibodies that cross-react with other grasses? We found that none of ten non-Bermuda grass extracts attained 50% inhibition of RAST binding between sera from patients in Arizona and Bermuda-coupled disks whereas Giant Bermuda grass extract was completely cross-reactive with Common Bermuda grass.

Binding Sites, Antibody↗
PubMed1987

Double-blind, placebo-controlled immunotherapy with mixed grass-pollen allergoids. I. Rush immunotherapy with allergoids and standardized orchard grass-pollen extract.

Forty-five grass pollen-allergic patients were randomly assigned to three groups according to their skin test and RAST sensitivities and the severity of seasonal rhinitis. Eleven patients were treated with placebo (group 1), 19 patients (group 2) were treated with a six-mixed grass-pollen allergoid prepared by mild formalinization with a two-step procedure, and 15 other patients were treated with a standardized orchard grass-pollen extract (group 3). Because of a different immunotherapy schedule, only patients placed in groups 1 and 2 received the extracts in a double-blind fashion. Rush immunotherapy was performed in 3 to 6 days, and the maintenance dose was subsequently administered weekly for 4 weeks and every 2 weeks until the end of the grass-pollen season. During the season, a coseasonal treatment was administered. Systemic reactions occurred during the rush protocol in 36.8% of patients treated with allergoid and 20% of patients who received the standardized extract. Only patients treated with allergoid had systemic reactions during maintenance dose. The reactions observed with the standardized extract were more severe. Total doses of allergoid ranged from 2350 to 13,500 protein nitrogen units. Symptoms and medication scores during the peak of the season were analyzed. Patients treated with the standardized allergen had a significant reduction of the number of days of symptoms during the month of June (9.5 +/- 6.7 days; p less than 0.005) and of medication scores (1.3 +/- 1.4; p less than 0.01) compared to patients receiving placebo (19.4 +/- 8.1 days; medication score, 2.8 +/- 2.1).(ABSTRACT TRUNCATED AT 250 WORDS)

Allergens↗
PubMed1993

Characterization of grass group I allergens in timothy grass pollen.

Using Phl p V-depleted timothy grass pollen extract (Phleum pratense) as immunogen, we obtained a monoclonal antibody, QG 4, which recognized proteins of 33, 35, and 37 kd as determined by Western blotting. The antibody cross-reacted with pollen proteins of other grass species in the molecular weight range of 30 to 37 kd. By means of two-dimensional polyacrylamide gel electrophoresis blot of timothy grass pollen extract, we demonstrated at least seven protein spots: two of 37 kd with isoelectric points of 6.4 and 6.6; four of 35 kd with isoelectric points of 6.5, 6.8, 7.1, and 7.3; and one of 33 kd with an isoelectric point of 8.5. These protein spots were also detected by patients' pooled serum. Microsequencing of the 20 N-terminal amino acid residues revealed structures with sequence identities up to 90% to the well-established allergen, Lol p I of ryegrass (Lolium perenne). Therefore we assume that the monoclonal antibody QG 4 recognized the corresponding allergen Phl p I in timothy grass pollen.

Allergens↗