Grass Pollen Allergens: Antigenic Relationships Detected Using Monoclonal Antibodies and Dot Blotting Immunoassay

Abstract

Monoclonal antibodies against major rye-grass pollen allergens have been used to detect cross-reactive determinants in other grass pollen extracts. Antibody binding was detected by the dot blotting immunoassay in which alkaline phosphatase-conjugated anti-mouse IgG was used as secondary antibody. Taxonomically ordered variations were found between pollen of 22 grass species representing all major natural groups. One of the antibodies, that bound to the 28 to 30Kd allergen, showed high species specificity; immunoblotting showed binding to similar polypeptides in Festuca elatior but to none of the other grasses tested. This simple assay has applications in standardizing grass pollen extracts.

IgE cross-reactivity between Lolium multiflorum and commercial grass pollen allergen extracts in Brazilian patients with pollinosis

“…Concerning the IgE-binding Lm components, immunoblot results revealed a number of immunodominant and minor allergenic bands recognized by SAR patients. The major components recognized by IgE (>90% recognition) had apparent molecular masses of 26, 28-30, and 32-35 kDa, the latter two probably corresponding to allergens from groups 1 and 5, since group 1 consists of glycoproteins of 32-35 kDa, and group 5 of non-glycosylated proteins with 28-30 kDa (26,27). In the Lm extract, antigenic components of 55 and 60-66 kDa were also recognized by IgE antibodies of most patients with pollinosis.…”

Isolation of cDNA encoding a newly identified major allergenic protein of rye-grass pollen: intracellular targeting to the amyloplast.

“…Proteins from the leaf, root, and seed were extracted by grinding the tissue in 150 mM NaCl/10 mM phosphate, pH 7.2/1 mM phenylmethylsulfonyl fluoride. Conditions for electrophoresis and immunoblotting were essentially as described (12). Monoclonal antibodies (mAbs) 3.2 and 21.3 bind specifically to the Lol pI group of antigens (9); mAb 40.1, although originating in a different laboratory, shows similar specificity (12,15).…”

“…Conditions for electrophoresis and immunoblotting were essentially as described (12). Monoclonal antibodies (mAbs) 3.2 and 21.3 bind specifically to the Lol pI group of antigens (9); mAb 40.1, although originating in a different laboratory, shows similar specificity (12,15). mAb 12.3 binds to a 31-kDa component in the Lol pI group (15).…”

“…Elevated levels of IgE specific for this allergen have been detected in sera of up to 95% of grass pollenallergic individuals (8). Antigenically cross-reacting proteins corresponding to Lol pI, known as the group I allergens, are present in the pollen of other clinically important grasses (9)(10)(11)(12).…”

Detection of Some Airborne Pollen Grain and Fungal Spore Allergens

“…The purpose of this study is to identify the IgEbinding allergens of some common airborne pollen grains (ragweed, tag alder, timothy grass and white ash) and fungi (Alternuria and Cladosporhm) species, which are widespread in temperate regions and significant causes of allergic disorder (Haas et al 1986, Baldo 1983, Singh & Knox 1985, Nilsson & Aas 1976, Gravesen 1979, Beaumont et al 1984. The protein blotting approach for the identification of important allergens allowed us to rapidly identify the major IgE-binding components in the above extracts.…”