Major allergen Phl p Va (timothy grass) bears at least two different IgE-reactive epitopes.
It is established that most grass pollen allergens consist of several isoforms of which the function is mainly still unknown. A number of these allergens belonging to group V have been cloned, sequenced, and expressed. Antigenic sites and IgE-reactive epitopes of the major allergen Phl p Va, are unknown. We have identified the complete cDNA sequence of a Phl p Va isoallergen by immunoscreening of a timothy grass pollen cDNA library and mixed oligonucleotide primed amplification of N-terminal cDNA. Additionally, we found an incomplete isoallergenic cDNA clone of the same protein. Immunoreactivity of the fusion proteins with patients' sera and monoclonal antibodies showed that the clones represent group Va allergens. Comparison of deduced amino acid sequences with published sequences of Lol p V and Poa p IX revealed a homology of 81.1% and 86.9%, respectively. With affinity-purified IgE antibodies recognizing the recombinant fusion protein, we can demonstrate the existence of a common group V IgE-reactive epitope. By construction of both an N-terminal and a C-terminal peptide of the complete Phl p Va and cross-inhibition, we identified at least two different IgE epitopes. Eleven patients showed variable IgE immunoreactivities to both IgE-reactive epitopes.
Bufe
A
A
Forschungsinstitut Borstel, Germany.
Becker
W M
WM
Schramm
G
G
Petersen
A
A
Mamat
U
U
Schlaak
M
M
D000485:Allergens / Q000235:genetics / Q000276:immunology*; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western; D004247:DNA / Q000032:analysis; D017931:DNA Primers; D000939:Epitopes / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D017343:Genes, Plant; D006801:Humans; D006969:Hypersensitivity, Immediate / Q000276:immunology; D007073:Immunoglobulin E / Q000276:immunology*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology*; D006109:Poaceae / Q000276:immunology*; D011058:Pollen / Q000235:genetics / Q000276:immunology*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014162:Transfection
D000485:Allergens; D000911:Antibodies, Monoclonal; D017931:DNA Primers; D000939:Epitopes; C076049:Phl p V protein, Phleum pratense; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E; D004247:DNA