Sequence polymorphisms and antibody binding to the group 2 dust mite allergens.
BACKGROUND
The group 2 allergens Der p 2, Der f 2 and Eur m 2 are 14-kD proteins with > 80% sequence identity. Isoforms within each genus have been identified which differ by 3-4 amino acids. The aim of this study was to investigate the importance of these substitutions to antibody binding.


METHODS
Recombinant allergens were expressed and purified from Escherichia coli. ELISA and skin testing were used to evaluate antibody binding. Molecular modeling of the tertiary structure was preformed to examine the location of substitutions.


RESULTS
The three Der f 2 isoforms and two of three of the Der p 2 isoforms reacted with all monoclonal antibodies (mAb). Der p 2.0101, the isoform with aspartate at position 114, bound all mAb except 1D8. Substitution of asparagine for aspartate restored binding of rDer p 2.0101 to mAb 1D8 and increased the correlation coefficient for IgE binding from 0.72 to 0.77. The three Der p 2 isoforms showed comparable skin test reactivity to nDer p 2 and commercial extract. rEur m 2.0101 bound to all mAb except 7A1 and when compared with rDer p 2 for IgE binding, r(2) = of 0.58 (n = 72). Lep d 2 did not react with mAb or with Dermatophagoides spp. allergic sera. Modeling revealed that Eur m 2, Lep d 2 and Tyr p 2 retain the tertiary fold of Der p 2 and the substitutions are on the surface.


CONCLUSIONS
mAb could distinguish isoform substitutions. IgE binding showed a good correlation among all isoforms, thus the recombinant allergens are useful for diagnosis.
Smith
A M
AM
Asthma and Allergic Diseases Center, University of Virginia, Charlottesville, VA 22908-1355, USA. ams3k@virginia.edu
Benjamin
D C
DC
Derewenda
U
U
Smith
W A
WA
Thomas
W R
WR
Chapman
M D
MD
D000485:Allergens / Q000235:genetics / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D039741:Antigens, Dermatophagoides; D006023:Glycoproteins / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D016297:Mutagenesis, Site-Directed; D011110:Polymorphism, Genetic; D020033:Protein Isoforms / Q000235:genetics / Q000276:immunology
D000485:Allergens; D000911:Antibodies, Monoclonal; D039741:Antigens, Dermatophagoides; D006023:Glycoproteins; D020033:Protein Isoforms; D007073:Immunoglobulin E