BACKGROUND: Paper mulberry (Morus papyrifera) pollen is considered to be one of the most clinically relevant aeroallergens in Pakistan. To date, the allergenicity of the pollen has not been investigated. OBJECTIVE: To characterize the sensitization profile of mulberry-allergic patients and the proteins of paper mulberry pollen contributing to pollinosis in the Pakistani population. METHODS: Proteins were extracted from mulberry pollen using different protocols. Immunoglobulin (Ig) E binding proteins to mulberry pollen was determined by ImmunoCAP testing and immunoblotting using sera from 29 mulberry pollen-allergic patients with positive skin prick test results to mulberry pollen antigens. The histamine release assay was performed in vitro to determine the allergenic potency of pollen extracts and a partially purified mulberry pollen allergen. The protein was identified using N-terminal sequencing and matrix-assisted laser desorption/ionization-time of flight spectrometry (MALDI-TOF/TOF). RESULTS: IgE sensitization to mulberry pollen was confirmed by positive ImmunoCAP results to pollen from Morus alba (white mulberry) in 23 out of 29 mulberry pollen-allergic patients. A 10-kDa protein from the paper mulberry pollen extract was considered a major allergen, along with additional IgE-reactive proteins. Sera from 79% of the patients reacted to the 10-kDa allergen, which showed substantial capacity to trigger histamine release in 3 out of 4 patients. N-terminal sequencing and MALDI-TOF/TOF yielded an amino acid sequence with no homology to known proteins. CONCLUSIONS: Mulberry-allergic patients are sensitized to multiple mulberry pollen allergens. We identified a novel 10-kDa protein that was a major allergen and should be further investigated for diagnostic and therapeutic purposes.
BACKGROUND:Paper mulberry (Morus papyrifera) pollen is considered to be one of the most clinically relevant aeroallergens in Pakistan. To date, the allergenicity of the pollen has not been investigated. OBJECTIVE: To characterize the sensitization profile of mulberry-allergicpatients and the proteins of paper mulberry pollen contributing to pollinosis in the Pakistani population. METHODS: Proteins were extracted from mulberry pollen using different protocols. Immunoglobulin (Ig) E binding proteins to mulberry pollen was determined by ImmunoCAP testing and immunoblotting using sera from 29 mulberry pollen-allergicpatients with positive skin prick test results to mulberry pollen antigens. The histamine release assay was performed in vitro to determine the allergenic potency of pollen extracts and a partially purified mulberry pollen allergen. The protein was identified using N-terminal sequencing and matrix-assisted laser desorption/ionization-time of flight spectrometry (MALDI-TOF/TOF). RESULTS: IgE sensitization to mulberry pollen was confirmed by positive ImmunoCAP results to pollen from Morus alba (white mulberry) in 23 out of 29 mulberry pollen-allergicpatients. A 10-kDa protein from the paper mulberry pollen extract was considered a major allergen, along with additional IgE-reactive proteins. Sera from 79% of the patients reacted to the 10-kDa allergen, which showed substantial capacity to trigger histamine release in 3 out of 4 patients. N-terminal sequencing and MALDI-TOF/TOF yielded an amino acid sequence with no homology to known proteins. CONCLUSIONS:Mulberry-allergicpatients are sensitized to multiple mulberry pollen allergens. We identified a novel 10-kDa protein that was a major allergen and should be further investigated for diagnostic and therapeutic purposes.