Togawa Akihisa, Panzani Raphael C, Garza Maritza A, Kishikawa Reiko, Goldblum Randall M, Midoro-Horiuti Terumi
Department of Pediatrics, Child Health Research Center, University of Texas Medical Branch, Galveston 77555-0366, USA.
Ann Allergy Asthma Immunol. 2006 Sep;97(3):336-42. doi: 10.1016/S1081-1206(10)60798-3.
The prevalence of seasonal allergic diseases of the upper airways is increasing in industrialized countries. The Cupressaceae are important causes of pollinosis, particularly in Europe.
To determine whether the pollen from Cupressus sempervirens (Italian cypress) contains a pathogenesis-related group 5 (PR-5) protein, similar to that found in other allergenic Cupressaceae pollens.
Messenger RNA was purified from Italian cypress pollen, and complementary DNA (cDNA) was synthesized. cDNAs for PR-5 proteins were amplified by polymerase chain reaction and extended by rapid amplification of cDNA ends methods. Recombinant Cup s 3 was expressed in Escherichia coli as a fusion protein. Inhibition enzyme-linked immunosorbent assays were used to test the allergenicity of Cup s 3.
Three cDNAs were cloned. These clones had approximately 95% identity to Jun a 3 and Cup a 3. Recombinant Cup s 3.0102 maltose-binding protein inhibited the IgE from most patients from binding to an extract of Italian cypress. The extent of inhibition suggested that antibodies to Cup s 3 were a prominent component of the IgE response to Italian cypress pollen.
Cup s 3, an allergen of Italian cypress pollen, was identified based on cross-reactivity and homology with other pollen PR-5 proteins, despite an apparently low level of protein expression. Variations in the content of Cup s 3 in the pollen from different regions or trees should be considered in the choice of extracts for diagnosis and specific immunotherapy for Italian cypress pollen hypersensitivity.
在工业化国家，上呼吸道季节性过敏性疾病的患病率正在上升。柏科植物是花粉症的重要病因，尤其是在欧洲。
确定地中海柏木（意大利柏）花粉中是否含有与其他致敏柏科花粉中发现的类似的病程相关蛋白5（PR-5）。
从意大利柏花粉中纯化信使核糖核酸，并合成互补脱氧核糖核酸（cDNA）。通过聚合酶链反应扩增PR-5蛋白的cDNA，并采用cDNA末端快速扩增法进行延伸。重组Cup s 3在大肠杆菌中作为融合蛋白表达。采用抑制酶联免疫吸附测定法检测Cup s 3的致敏性。
克隆了三个cDNA。这些克隆与Jun a 3和Cup a 3具有约95%的同一性。重组Cup s 3.0102麦芽糖结合蛋白抑制了大多数患者的免疫球蛋白E与意大利柏提取物的结合。抑制程度表明，针对Cup s 3的抗体是对意大利柏花粉免疫球蛋白E反应的一个主要成分。
尽管蛋白表达水平明显较低，但基于与其他花粉PR-5蛋白的交叉反应性和同源性，鉴定出了意大利柏花粉的一种变应原Cup s 3。在选择用于意大利柏花粉过敏症诊断和特异性免疫治疗的提取物时，应考虑不同地区或树木的花粉中Cup s 3含量的差异。
