家蚕30kD脂蛋白家族的过敏原性及其致敏机制研究

Silkworm alllery is a domestic occupational as well as food allery with Chinese characteristics. However, its major allergens have not been identified. Our previous work found that 30 kD lipoprotein family of silkworm were the most important allergenic components and demonstrated potent allergenicity. We herein will illuminate two scientific problems: (1)we will evaluate the allergenicity of silkworm 30 kD lipoprotein family and submit the family to the WHO/IUIS Allergen Nomenclature Sub-committee for systematic allergen nomenclature.(2)we will conduct research to figure out the mechanism of silkworm 30 kD lipoprotein family-aroused silkworm allergies. To achieve these goals we will carry out the following experiments: (1)we will purify native as well as recombinant 30 kD lipoprotein family members of silkworm and explore the allergenicity of these proteins; we will investigate the immuno-reactivity of the 30 kD lipoprotein family members and observe the effects of various physical, chemical as well biological treatments on the allergenicity of the family members with high allergenicity; We will perform systematic allergen nomenclature and get official approval of the WHO/IUIS Allergen Nomenclature Sub-committee. (2)We will probe IgE epitopes of 30 kD family members with high allergenicity; we will compare the secondary structures of the protein members and look into the correlation between proteins structures and allergenicity; we will crystallize the family members with high allergenicity and analyze the mechanism of lipoprotein-caused silkworm allergies. Our project focuses on an allergic disease with distinguish Chinese Characteristics and will lay the foundation for future diagnosis and therapy of silkworm allergies.

家蚕过敏是一种中国特色的职业性过敏与食物过敏，其主要过敏原成分一直没有确定,机制也未见研究。我们首先发现家蚕30kD 脂蛋白家族具有较强的过敏原活性而且是家蚕最主要的过敏原成分。本项目中我们将研究两个科学问题：（1）家蚕30kD脂蛋白家族的过敏原性高低如何并进行过敏原的标准命名；（2）研究30kD脂蛋白的致敏机制。研究内容包括：（1）30kD脂蛋白家族天然及重组蛋白的纯化；各家族成员与过敏病人IgE 的反应率及反应强度的高低；比较高过敏原性与低过敏原性家族成员的免疫反应性；研究各种物理、化学与生物因素对高过敏性成员过敏原性的影响；进行过敏原的标准命名并获得国际过敏原命名委员会的批准。（2）检测高过敏原个体的线性IgE 表位。比较各个成员的二级结构，分析其与过敏原性的联系。对高致敏性成员进行蛋白质结晶，结合其分子结构研究其致敏机制。本研究具有鲜明的中国特色，为家蚕过敏的诊断与治疗打下基础。

我们开展了儿童家蚕过敏反应过敏原图谱的研究，完成了家蚕30kD家族成员的9个家族成员的克隆、表达与纯化，鉴定了BMLP7的IgE反应性。我们以霍乱毒素为佐剂建立了蚕蛹过敏原的食物过敏模型。建立的模型表现出了明显的食物过敏症状。我们开展了家蚕过敏的机制研究。用不同浓度的蚕蛾过敏原处理上皮细胞系A549。发现，不同浓度的蚕蛾过敏原对A549的生长没有明显致死或者促生长作用。用蛋白处理A549细胞24小时后，用RT-PCR检测了A549细胞中重要的过敏相关分子（如TSLP、IL-33等）的表达水平变化，取得了重要发现，发现了一系列Th2型的细胞因子用明显的上调。我们根据以下的发现，提出了重要的假说，并在后续实验中取得了重要的实际进展，将在后续研究中给予体现。我们的研究为研究家蚕过敏的诊断与治疗打下了基础。