Study of the epitope structure of purified Dac G I and Lol p I, the major allergens of Dactylis glomerata and Lolium perenne pollens, using monoclonal antibodies.
The use of mAb allowed us to further analyze the cross-reactivity between purified Dac g I and Lol p I, the major allergens of Dactylis glomerata (cocksfoot) and Lolium perenne (Rye grass), respectively. It was first shown, using IEF, followed by immunoprinting, that serum IgE antibodies from most grass-sensitive patients recognize both Dac g I and Lol p I. Second, three different anti-Lol p I mAb, 290A-167, 348A-6, and 539A-6, and one anti-Dac g I mAb, P3B2 were all shown to react with Dac g I and Lol p I, indicating that the two molecules share common epitopes. Epitope specificity of the mAb was determined by competitive binding inhibition of a given labeled mAb to solid phase fixed Dac g I or Lol p I by the mAb. The results indicated that the four mAb are directed against four different and non-overlapping epitopes present on both allergens. Using double-binding RIA, our data strongly suggest that the common epitopes are not repetitive on both molecules. In addition to their similar physicochemical characteristics, such as isolectric points and m.w., Dac g I and Lol p I share four identical epitopes. Binding inhibition of human IgE to Lol p I and Dac g I by the mAb was also assessed. The results indicated that each mAb was able to inhibit such reactions to variable degree but no additive inhibition was observed when two mAb of different specificities were used in combination, suggesting that the human IgE binding site is partially shared by each epitope recognized by the four mAb.
Mourad
W
W
Unité de Recherche en Inflammation et Immunologie-Rhumatologie, CHUL, Québec, Canada.
Mécheri
S
S
Peltre
G
G
David
B
B
Hébert
J
J
D000485:Allergens / Q000276:immunology / Q000302:isolation & purification*; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D052179:Antigens, Plant; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D003429:Cross Reactions; D000939:Epitopes / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010940:Plant Proteins; D006109:Poaceae; D011058:Pollen / Q000276:immunology / Q000302:isolation & purification*; D006255:Rhinitis, Allergic, Seasonal / Q000276:immunology; D012434:Secale
D000485:Allergens; D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; C057603:Dac g I protein, Dactylis glomerata; D000939:Epitopes; C042814:Lol p I protein, Lolium perenne; D010940:Plant Proteins; D007073:Immunoglobulin E