Purification and immunological characterization of a 12-kDa allergen from Epicoccum purpurascens

Exposure to Epicoccum purpurascens is implicated in respiratory allergies and asthma. Several allergens of clinical importance were identified in Epicoccum extract (EE), but only one allergen has been isolated and characterized. In the present study, a 12-kDa allergen was isolated from an Epicoccum spore-mycelial extract by concanavalin-A sepharose, reverse-phase hydrophobic and gel filtration chromatography. The purified protein was recognized as a single 12-kDa allergen on immunoblot with a serum pool of Epicoccum- sensitive patients. Of the 94 respiratory allergy patients tested intradermally, 17 showed marked positive skin reactions to EE and 12 of them reacted with the 12-kDa protein, indicating a diagnostic sensitivity of 70%. More than 80% patients' sera showed immunoglobulin E (IgE) reactivity to the purified protein in enzyme-linked immunosorbent assay and immunoblot, identifying it as a major allergen. Preincubation of pooled serum with the protein led to inhibition of IgE binding to solid-phase-bound EE (effective concentration 50%=180 ng). Twelve of the 17 serum samples showed significant basophil histamine release upon stimulation with purified protein. The protein induced significant proliferation of peripheral blood mononuclear cells of 13 patients. A high level of interleukin-4 in the culture supernatant of these cells indicated induction of a T-helper type 2 response. The purified 12-kDa protein is a clinically relevant allergen and has potential for the diagnosis and therapy of Epicoccum allergies.     

Identification, expression and characterisation of a major salivary allergen (Cul s 1) of the biting midge Culicoides sonorensis relevant for summer eczema in horses

Salivary proteins of Culicoides biting midges are thought to play a key role in summer eczema (SE), a seasonal recurrent allergic dermatitis in horses. The present study describes the identification, expression and clinical relevance of a candidate allergen of the North American midge Culicoides sonorensis. Immunoblot analysis of midge saliva revealed a 66 kDa protein (Cul s 1) that was bound by IgE from several SE-affected (SE+) horses. Further characterisation by fragmentation, mass spectrometry and bioinformatics identified Cul s 1 as maltase, an enzyme involved in sugar meal digestion. A cDNA encoding Cul s 1 was isolated and expressed as a polyhistidine-tagged fusion protein in a baculovirus/insect cell expression system. The clinical relevance of the affinity-purified recombinant Cul s 1 (rCul s 1) was investigated by immunoblotting, histamine release testing (HRT) and intradermal testing (IDT) in eight SE+ and eight control horses. Seven SE+ horses had rCul s 1-specific IgE, whereas only one control animal had IgE directed against this allergen. Furthermore, the HRT showed rCul s 1 induced basophil degranulation in samples from seven of eight SE+ horses but in none of the control animals. rCul s 1 also induced immediate (7/8), late-phase (8/8) and delayed (1/8) skin reactivity in IDT on all SE+ horses that had a positive test with the whole body extract (WBE) of C. sonorensis. None of the control horses showed immediate or delayed skin reactivity with rCul s 1, and only one control horse had a positive late-phase response, while several non-specific late-phase reactions were observed with the insect WBE. Thus, we believe rCul s 1 is the first specific salivary allergen of C. sonorensis to be described that promises to advance both in vitro and in vivo diagnosis and may contribute to the development of immunotherapy for SE in horses.     

牛奶过敏原β-乳球蛋白的重组制备及磁微粒化学发光检测法的建立

运用原核系统表达牛奶β-乳球蛋白(Bos d5)蛋白,建立一种纳米磁微粒化学发光方法用于检测牛奶组分过敏原β-乳球蛋白特异性Ig E抗体的含量。通过优化合成牛奶β-乳球蛋白基因,与质粒重组导入Rosetta原核表达菌株中表达目标蛋白,纯化的重组蛋白采用生物素标记后,在化学发光平台上进行过敏原项目检测。获得纯度﹥85%的22.5 kD重组牛β-乳球蛋白,将生物素化的重组蛋白用于牛奶β-乳球蛋白纳米磁微粒检测试剂盒,检测临床110例血清样本,和Phadia进行方法学比对阳性符合率为88.9%,阴性符合率97.3%,总符合率94.5%,P<0.001,χ^2=84.238,Kappa=0.874,其与Phadia参照试剂盒同样具有良好的检测能力。原核表达系统中获得的重组牛奶过敏组分β-乳球蛋白具有较好的免疫活性,开发的免疫诊断试剂盒具备良好的性能,可用于临床辅助诊断。     

5岁以下哮喘患儿血清过敏原病例对照研究及相关性分析

摘要 目的：探讨5岁以下哮喘儿童与血清特异性过敏原（specific IgE,sIgE）的分布情况。方法：本研究采用免疫印迹法对 2019 年1月至 2019 年12月在西安交通大学第二附属医院住院的5岁以下62例哮喘患儿和49例喘息患儿的行血清特异性过敏原检测,对比分析5岁以下哮喘和喘息儿童过敏原分布情况及与哮喘的发病关系。结果：户尘螨、猫毛皮屑、狗毛皮屑、蒿草、葎草、桤杨柳山毛榉橡胡桃、烟曲霉、念珠菌点青霉分枝孢霉交链孢霉黑曲霉吸入过敏原和花生黄豆、腰果开心果榛子杏仁核桃、虾蟹、桃苹果芒果荔枝草莓食物过敏原这12类过敏原在哮喘组与喘息组有显著差异（P<0.05）,与哮喘发病有关。多因素logistic回归分析结果显示户尘螨、猫毛皮屑、坚果类、霉菌、水果类是哮喘发病的危险因素（P<0.05）。户尘螨、猫毛皮屑和虾蟹是男性哮喘患儿发病的危险因素,念珠菌点青霉分枝孢霉交链孢霉黑曲霉是女性哮喘患儿发病的危险因素（P<0.05）。结论：血清特异性（sIgE）过敏原在哮喘与喘息患儿中分布不同,同时发现过敏原在哮喘患儿中存在性别差异,故对哮喘患儿进行过敏性检测可以作为回避过敏原的依据。     

One-Year pollen and spore calendars of Saudi Arabia Al-Khobar, Abha and Hofuf

Airborne Pollen grains and Spores of three different cities viz., Al-Khobar (1987–1988), Abha (1991–1992) and Hofuf (1992–1993) in Saudi Arabia were studied using Burkard Volumetric Seven-Day Spore Trap. The data were analyzed in relation to their allergenic capability and one-year pollen and spore calendars were designed to correlate the patients’ symptoms as well as for selection of appropriate allergen extracts for diagnosis and treatment of allergic diseases. Amongst pollen group, Amaranthus viridis, Plantago spp., Chenopodium album, Ricinus communis, Rumex vesicarius, Juniperus spp., Parkinsonia aculeata, Prosopis spp., and Phoenix dactylifera were some of the frequent types. Amongst the fungal spores group Cladosporium, Smuts spores, Colored basidiospores, Alternaria, Ulocladium and Drechslera were the dominant types.     

The ecology of microscopic life in household dust

We spend the majority of our lives indoors; yet, we currently lack a comprehensive understanding of how the microbial communities found in homes vary across broad geographical regions and what factors are most important in shaping the types of microorganisms found inside homes. Here, we investigated the fungal and bacterial communities found in settled dust collected from inside and outside approximately 1200 homes located across the continental US, homes that represent a broad range of home designs and span many climatic zones. Indoor and outdoor dust samples harboured distinct microbial communities, but these differences were larger for bacteria than for fungi with most indoor fungi originating outside the home. Indoor fungal communities and the distribution of potential allergens varied predictably across climate and geographical regions; where you live determines what fungi live with you inside your home. By contrast, bacterial communities in indoor dust were more strongly influenced by the number and types of occupants living in the homes. In particular, the female : male ratio and whether a house had pets had a significant influence on the types of bacteria found inside our homes highlighting that who you live with determines what bacteria are found inside your home.