Investigations of the allergens of cocksfoot grass (dactylis glomerata) pollen

Calam, D.H.; Davidson, J.; Ford, Anna

doi:10.1016/s0021-9673(01)90903-3

Cited by 15 publications

(3 citation statements)

References 7 publications

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“…The subject of this study is the pollen of orchard grass (cocksfoot) (Dactylis glomerata), a species which is widespread in temperate regions and a significant cause of allergic disease [3,6,7], Preliminary investi gations have shown that extracts of this species con tain a number of allergens [8][9][10][11][12] but none of the al lergens have yet been purified, characterized and as sessed for biological potency and the full spectrum of allergens has yet to be revealed.…”

Section: Introductionmentioning

confidence: 99%

Identification of Orchard Grass <i>(Dactylis glomerata)</i> Pollen Allergens following Electrophoretic Transfer to Nitrocellulose

Ford¹,

Tovey²,

Baldo³

1985

Int Arch Allergy Immunol

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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 > 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.

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Section: Introductionmentioning

confidence: 99%

Identification of Orchard Grass <i>(Dactylis glomerata)</i> Pollen Allergens following Electrophoretic Transfer to Nitrocellulose

Ford¹,

Tovey²,

Baldo³

1985

Int Arch Allergy Immunol

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“…4,550,085). Then 50 pi of prepared blood samples was incubated with a 20-pl dilution of lyophilized polypeptides (10,11,12,14,16,19,23,26,28,30,35 and 45 kD) or crude extract of timothy grass pollen (2 mg/ml in Pipes-AMC; dilution series of 10°, 10"', I0"2 and 10'3 were tested) and added to the microtitre plates. After incuba tion, released histamine was bound to the microfibres.…”

Section: Histamine Release Testmentioning

confidence: 99%

“…Purification of single timothy grass pollen components was a prerequisite. Many different technical approaches have been made to isolate grass pollen allergens [5,7,10,[15][16][17], We isolated allergenic polypeptides of timothy grass pollen extract by means of preparative sodium do decyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) [18,19], These polypeptides were used in inhibition experiments. Based on the results of the inhibitions, it became obvious that allergens of timo thy grass detected in immunoblot dispose of private and cross-reactive determinants for IgE binding.…”

Section: Introductionmentioning

confidence: 99%

Hyposensitization Therapy with Whole Pollen Extract or Purified Allergens Monitored by Immunoblotting

Jarolim

Matthiesen²,

Skov

et al. 1990

Int Arch Allergy Immunol

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Patients allergic to grass pollen were hyposensitized with two major allergenic components or whole extract of timothy grass pollen. Specific IgE, IgG1, and IgG4 formed during immunotherapy were analyzed by immunoblotting. Similar antibody-binding patterns were observed in both patient groups. Inhibition experiments using allergenic components isolated by preparative sodium dodecyl sulphate-polyacrylamide gel electrophoresis indicated that all antigenic components of timothy grass pollen detected in immunoblot dispose of private and cross-reactive determinants for binding of human IgE. The worse clinical outcome of immunotherapy after hyposensitization with two cross-reactive components did, therefore, not correlate with a specific antibody formation pattern.

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Macromolecules

Harborne

1984

Phytochemical Methods

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Investigations of the allergens of cocksfoot grass (dactylis glomerata) pollen

Cited by 15 publications

References 7 publications

Identification of Orchard Grass <i>(Dactylis glomerata)</i> Pollen Allergens following Electrophoretic Transfer to Nitrocellulose

Identification of Orchard Grass <i>(Dactylis glomerata)</i> Pollen Allergens following Electrophoretic Transfer to Nitrocellulose

Hyposensitization Therapy with Whole Pollen Extract or Purified Allergens Monitored by Immunoblotting

Macromolecules

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