High-molecular-size mite allergens with high carbohydrate content in mite extract containing mite feces of Dermatophagoides farinae.

A series of mite allergens with high carbohydrate content was fractionated from mite extract containing mite feces by ultrafiltration, and successive chromatography on Ultrogel AcA 54, Sepharose CL-6B, and DEAE Toyopearl 650S. The final fractions isolated were termed HM1-2-Eff, -0.1A, -0.1B, -0.3A, -0.3B, and -1M according to the eluted salt concentration. These six fractions were stained weakly with Coomassie Brilliant Blue but strongly with periodate-Schiff's reagent on SDS-PAGE. Their molecular weight were estimated to be 150-155 KD on Sepharose CL-6B. All of these six fractions provoked strong erythema in skin of mite-sensitive and asthmatic patients. HM1-2-Eff, -0.1A, -0.1B and -0.3B reacted with 73% of mite-sensitive patients' IgE by the nitrocellulose dot blotting. One ng/ml of 4 fractions (HM1-2-0.1A, 0.3A, 0.3B and -1M) released histamine from mite allergic patient's blood cells. HM1-2-0.3A induced proliferation of T cell from immunized mouse by mite extract containing mite feces (Dff).