Dermatophagoides farinae Allergens Diversity Identification by Proteomics

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. Molecular & Cellular

Glycoproteomic Analysis of Seven Major Allergenic Proteins Reveals Novel Post-translational Modifications

“…32. These results clearly demonstrate that K 50 , and not T 53 or T 67 , is modified by Hex. Similar results were obtained for the Der f 2 allergen.…”

“…S8 and S9) both T 53 and T 67 , was observed without Hex modification at m/z 1595.79. In addition, the c4 fragment, which excludes both T 53 and T 67 but includes K 50 , was observed with a 162.05 amu mass increment, corresponding to a Hex residue, at m/z 609. 32.…”

“…The results therefore strongly suggest that these glycans on the allergens originate from the mites. HDM are believed to have a repertoire of up to 30 distinct allergens (50,51) and it is likely that the identified glycans may be present on many other allergens from mites. Glycations have previously been described on peanut allergens Ara h 1 and Ara h 3 as well as on ovalbumin (52,53).…”

Hierarchy and molecular properties of house dust mite allergens

“…71 The IgE responses have yet to be quantitated or compared with other allergens. 72 Allergenicity was further examined with chromatographically isolated proteins although the profiles show a requirement for further purification and then quantitation and the exclusion of cross-reactive carbohydrate binding. Little IgE binding to myosin alkali light-chain protein, pyrophosphatase and ferritin has been found to recombinant proteins 71,73 in other studies and ferritin has been found not to induce Th2-biased cytokine responses.…”

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“…Detection of cross-reactive pollen allergens: Previously, proteins of GST-family were identified as allergens in house dust mite allergen, Der 20 and German cockroach, Bla g5 21 . We investigated a potential cross-reactivity between GST from birch and GST from parthenium.…”