Abstract：
Background. Parthenium hysterophorus pollen induces chronic clinical conditions such as allergic rhinitis and bronchial asthma. Among the plethora of proteins in the pollens, only few were reported to induce allergy. Currently sensitization to P. hysterophorus pollen allergen is diagnosed by skin prick test (SPT) using the entire pollen extract instead of using the specific allergen. Methods. In P. hysterophorus sensitized patients, SPT was done using the crude pollen extract, 40 kDa allergenic pollen protein and two commercially synthesized allergen epitopes (17 and 24) of P. hysterophorus. Dot-blot of allergen epitopes was done using P. hysterophorus sensitized sera. Crude pollen extract (1, 1.25, 2.5, 5 and 10 µg/mL), 40 kDa allergenic protein (3 µg/mL), and allergen epitopes (3µg/mL) were used to perform Basophil Activation Test (BAT). Results. Crude pollen extract at 2.5, 5, 10 μg/mL and 40 kDa allergenic protein at 3μg/mL concentrations induced wheal and flare reaction by around 15 minutes, whereas commercially synthesized allergen epitopes at 3μg/mL induced wheal and flare reactions in less than 10 minutes. Allergen epitopes (3 µg/mL) revealed strong reactivity with sensitized patient\'s IgE in dot-blot analysis. Basophil activation Test using crude pollen extract (2.5, 5, 10 µg/mL), 40 kDa allergenic protein (3 µg/mL), and allergenic epitopes (3µg/mL) indicated significant basophil activation (as measured by CD63 expression) in sensitized patients. Conclusions. The 40 kDa allergenic protein and its allergenic epitopes (17 and 24) induced phenotypic and cellular immune responses in P. hysterophorus sensitized individuals. The tested allergenic epitopes (17 and 24) induced faster wheal and flare reactions in comparison with the crude extract and the 40 kDa allergenic protein. The novel 40kDa allergenic protein and its allergen epitopes identified here may be useful for the development of component-resolved diagnosis (CRD) while also serving as a potential therapeutic lead for desensitization treatment for P. hysterophorus pollen induced allergy.
摘要：
■。背景。Partheniumhysterophorphus花粉诱导慢性临床疾病，例如过敏性鼻炎和支气管哮喘。在花粉中过多的蛋白质中，据报道只有少数人诱发过敏。目前,通过皮肤点刺试验(SPT),使用整个花粉提取物而不是使用特定的过敏原来诊断对子宫磷花粉过敏原的致敏。方法。在P.hyperphorus致敏患者中，使用粗花粉提取物进行SPT，40kDa变应原花粉蛋白和两个商业合成变应原表位(17和24)。变应原表位的斑点印迹使用P.hysterophorus致敏血清进行。粗花粉提取物（1、1.25、2.5、5和10µg/mL），40kDa致敏蛋白（3µg/mL），和变应原表位(3μg/mL)用于进行嗜碱性粒细胞活化试验(BAT)。结果。2.5、5、10μg/mL的粗花粉提取物和3μg/mL浓度的40kDa变应原蛋白在约15分钟内诱导了风团和耀斑反应，而商业合成的3μg/mL变应原表位在不到10分钟的时间内诱导了风疹和耀斑反应。变应原表位(3μg/mL)在斑点印迹分析中显示与致敏患者IgE的强反应性。嗜碱性粒细胞活化试验，使用粗花粉提取物（2.5、5、10µg/mL），40kDa致敏蛋白（3µg/mL），和变应原性表位(3µg/mL)表明致敏患者中显著的嗜碱性粒细胞活化(通过CD63表达测量).Conclusions.40kDa变应原性蛋白及其变应原性表位(17和24)在P.hy致敏个体中诱导表型和细胞免疫应答。与粗提物和40kDa变应原蛋白相比，测试的变应原性表位（17和24）诱导了更快的风团和耀斑反应。本文鉴定的新型40kDa变应原性蛋白及其变应原表位可用于开发成分分辨诊断（CRD），同时也可作为对P.heserophorus花粉诱导的变态反应的脱敏治疗的潜在治疗线索。
