To characterize the allergens of Blomia tropicalis, a cDNA library was constructed and screened with allergic sera from asthmatic patients. One clone, Bt6, was subcloned and sequenced. The nucleotide sequence of 934 bp length shows a 390-bp reading frame which encodes a 130-amino acid protein with a MW 14.8 kD. No potential glycosylation site was found in the predicted protein. The inferred amino acid sequence has no homology to known allergens. It has a cytosolic fatty acid-binding protein (FABP) signature at 5-22 amino acid residues, 42.3% identity with the Sml4-FABP of Schistosoma mansoni and 36% identity with FABPs from rat, mouse, bovine and human. The protein was expressed as a GST fusion protein and the purified GST-BtG used for dot blot, RAST and RAST inhibition assays. The frequency of IgE binding of allergic sera to Bt6 was low (11%) and usually weak. One positive serum did, however, show strong reactivity by RAST and dot blot and Bt6 could inhibit 60% of the IgE binding of this serum to the B. tropicalis extract. These data show that Bt6 encodes a mite FABP with allergenic properties, which are pronounced in some atopic subjects.

The mite Blomia tropicalis is a potent source of allergens in tropical and subtropical regions. So far most of these allergens have only been studied by immunoblotting. To characterize them at the molecular level a lambda gt11 complementary DNA library was constructed from messenger RNA isolated from whole B. tropicalis mites. This library was screened by using pooled sera from patients allergic to B. tropicalis in a plaque IgE radioimmunoassay. A B. tropicalis IgE-positive clone (Bt-M) was selected for immunologic studies. After subcloning into pBluescript (Stratagene, La Jolla, Calif.), it produced a sequence of 310 bp, with a probable amino acid sequence of 72 residues for the expressed peptide. The recombinant protein was transferred to nitrocellulose filters and probed with 100 sera from patients allergic to B. tropicalis. Forty-seven percent of sera reacted with the recombinant allergen. Immunoblottings performed with allergic serum and B. tropicalis-affinity-purified IgE demonstrated that the recombinant protein shares allergenic epitopes with the 11-13, 14, and 16 kd native allergens of B. tropicalis, which are known to be important allergens of this mite.

A panel of 188 unrelated Caucasian subjects who were exposed to the larvae of Chironomus thummi (Diptera, nonbiting midges) was HLA-typed by polymerase chain reaction amplification of the second exons of the DRB, DQA1, and DQB1 genes followed by dot-blot hybridization with sequence-specific oligonucleotide probes. Type I sensitization to the allergen Chi t I and a large number of other inhalant allergens was determined by RAST and skin testing. Sixty-one individuals were found to be sensitized to Chi t I, of whom 24 were sensitive to this allergen and to no other allergens tested. Statistical analyses showed that only in the latter group were the HLA-D genes DRB1*0101, DQA1*0101, and DQB1*0501 associated with IgE-responsiveness to Chi t I. These results suggest that HLA associations responsiveness to certain allergens may be more striking in monosensitized subjects.