Caraballo L, Avjioglu A, Marrugo J, Puerta L, Marsh D
Institute of Immunological Research, University of Cartagena, Colombia.
J Allergy Clin Immunol. 1996 Sep;98(3):573-9. doi: 10.1016/s0091-6749(96)70091-x.
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.
热带果螨是热带和亚热带地区一种重要的过敏原来源。到目前为止,这些过敏原大多仅通过免疫印迹法进行研究。为了在分子水平上对它们进行表征,从整个热带果螨分离的信使核糖核酸构建了一个λgt11互补脱氧核糖核酸文库。该文库通过在噬菌斑免疫球蛋白E放射免疫测定中使用对热带果螨过敏患者的混合血清进行筛选。选择了一个热带果螨免疫球蛋白E阳性克隆(Bt-M)进行免疫学研究。亚克隆到pBluescript(Stratagene,拉霍亚,加利福尼亚州)后,它产生了一个310碱基对的序列,所表达肽的可能氨基酸序列为72个残基。将重组蛋白转移到硝酸纤维素滤膜上,并用100份对热带果螨过敏患者的血清进行检测。47%的血清与重组过敏原发生反应。用过敏血清和热带果螨亲和纯化的免疫球蛋白E进行的免疫印迹表明,重组蛋白与热带果螨的11 - 13、14和16千道尔顿天然过敏原共享过敏原表位,已知这些是该螨的重要过敏原。