Ball T, Linhart B, Sonneck K, Blatt K, Herrmann H, Valent P, Stoecklinger A, Lupinek C, Thalhamer J, Fedorov A A, Almo S C, Valenta R
Department of Pathophysiology, Center for Physiology, Pathophysiology and Immunology, Medical University of Vienna, Austria.
Allergy. 2009 Apr;64(4):569-80. doi: 10.1111/j.1398-9995.2008.01910.x. Epub 2009 Feb 19.
The major timothy grass pollen allergen, Phl p 1, resembles the allergenic epitopes of natural group I grass pollen allergens and is recognized by more than 95% of grass-pollen-allergic patients. Our objective was the construction, purification and immunologic characterization of a genetically modified derivative of the major timothy grass pollen allergen, Phl p 1 for immunotherapy of grass pollen allergy.
A mosaic protein was generated by PCR-based re-assembly and expression of four cDNAs coding for Phl p 1 fragments and compared to the Phl p 1 wild-type by circular dichroism analysis, immunoglobulin E (IgE)-binding capacity, basophil activation assays and enzyme-linked immunosorbent assay competition assays. Immune responses to the derivative were studied in BALB/c mice.
Grass-pollen-allergic patients exhibited greater than an 85% reduction in IgE reactivity to the mosaic as compared with the Phl p 1 allergen and basophil activation experiments confirmed the reduced allergenic activity of the mosaic. It also induced less Phl p 1-specific IgE antibodies than Phl p 1 upon immunization of mice. However, immunization of mice and rabbits with the mosaic induced IgG antibodies that inhibited patients' IgE-binding to the wild-type allergen and Phl p 1-induced degranulation of basophils.
We have developed a strategy based on rational molecular reassembly to convert one of the clinically most relevant allergens into a hypoallergenic derivative for allergy vaccination.
主要的梯牧草花粉过敏原Phl p 1与天然I组草花粉过敏原的致敏表位相似，超过95%的草花粉过敏患者可识别该过敏原。我们的目标是构建、纯化并对主要梯牧草花粉过敏原Phl p 1的转基因衍生物进行免疫学特性分析，用于草花粉过敏的免疫治疗。
通过基于PCR的重组和表达编码Phl p 1片段的四个cDNA生成一种嵌合蛋白，并通过圆二色性分析、免疫球蛋白E（IgE）结合能力、嗜碱性粒细胞活化试验和酶联免疫吸附试验竞争试验与Phl p 1野生型进行比较。在BALB/c小鼠中研究对该衍生物的免疫反应。
与Phl p 1过敏原相比，草花粉过敏患者对嵌合体的IgE反应性降低了85%以上，嗜碱性粒细胞活化实验证实了嵌合体的致敏活性降低。在小鼠免疫后，它诱导产生的Phl p 1特异性IgE抗体也比Phl p 1少。然而，用嵌合体免疫小鼠和兔子可诱导产生IgG抗体，该抗体可抑制患者IgE与野生型过敏原的结合以及Phl p 1诱导的嗜碱性粒细胞脱颗粒。
我们开发了一种基于合理分子重组的策略，将临床上最相关的过敏原之一转化为用于过敏疫苗接种的低致敏性衍生物。
Zotero 插件