Abstract
The most important indoor allergens for humans are house dust mites (HDM). Fourteen Dermatophagoides farinae allergens (Der f 1-3, 6, 7, 10, 11, 13-18, and 22) are reported although more than 30 allergens have been estimated in D. farinae. Seventeen allergens belonging to 12 different groups were identified by a procedure of proteomics combined with two-dimensional immunoblotting from D. farina extracts. Their sequences were determined by Edman degradation, mass spectrometry analysis, and cDNA cloning. Their allergenicities were assayed by enzyme-linked immunosorbent assay inhibition tests, immunoblots, basophil activation test, and skin prick tests. Eight of them are the first report as D. farinae allergens. The procedure of using a proteomic approach combined with a purely discovery approach using sera of patients with broad IgE reactivity profiles to mite allergens was an effective method to investigate a more complete repertoire of D. farinae allergens. The identification of eight new D. farinae allergens will be helpful for HDM allergy diagnosis and therapy, especially for patients without response for HDM major allergens. In addition, the current work significantly extendedthe repertoire of D. farinae allergens.
Highlights
The house dust mites (HDM)1 are major sources of indoor allergens for humans, which induce asthma, rhinitis, dermatitis, and other allergic diseases [1]
Allergens Purification from D. farinae Mite Extracts (DME)—As indicated in Fig. 1A, the supernatant of dust mite extract (DME) was divided into four fractions by Sephadex G-75 gel filtration
Their primary structures and allergenicities were characterized by Edman degradation, cDNA cloning, mass spectrometry, competitive Enzyme-linked Immunosorbent Assay (ELISA), Basophil Activation Test (BAT), and Skin Prick Test (SPT)
Summary
The house dust mites (HDM) are major sources of indoor allergens for humans, which induce asthma, rhinitis, dermatitis, and other allergic diseases [1]. It has been found that there are many other HDM allergens containing high IgE binding activity these are present in low and variable concentrations in mite extracts (minor allergens), usually at less than 1% of the group 1 and 2 allergens [3]. Allergens present in low amount in mite extracts, which can induce high titers of IgE, suggest that they are potent at low concentration. Another possibility is that the amount of allergen required to induce allergic responses in the airways is more than that required to induce IgE. 17 allergens or their isoforms have been identified from the mite extracts of D. farinae by a procedure of proteomics combined with two-dimensional immunoblotting. Eight of them are the first to be reported as mite allergens
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