Ghosal Kavita, Saha Bodhisattwa, Gupta Bhattacharya Swati
Division of Plant Biology, Bose Institute, Main Campus, 93/1, A.P.C. Road, Kolkata, West Bengal 700009 India.
Allergy Asthma Clin Immunol. 2016 Jul 28;12:33. doi: 10.1186/s13223-016-0135-z. eCollection 2016.
The incidence of allergic diseases is increasing gradually and is a global burden affecting the socio-economic quality of life. Identification of allergens is the first step towards paving the way for therapeutic interventions against atopic diseases. Our previous investigation figured out that total pollen load correlated significantly with the rise of respiratory allergy in a subtropical city in India. The most dominant pollen responsible for IgE sensitivity in most patients emerged to be from Lantana camara (LC) an obnoxious weed growing in and around suburban areas of West Bengal. In this study, we identified allergenic components from this shrub using an immunoproteomic approach.
Determination of dominant pollen species was done using aerobiological sampling during two consecutive years and correlated with hospitalization and skin prick test. Serum was collected from LC positive patients and checked for in vitro allergenicity using ELISA and Histamine assay. Total proteome was profiled in SDS-PAGE, 2D PAGE and immunoblotted to detect IgE binding proteins which were further identified using mass spectrometry.
Lantana camara pollen emerged as a significant contributor from the correlation study with hospital admission of the respiratory allergy sufferers and its extract demonstrated an elevated IgE response in ELISA and histamine release assay tests. Five IgE reactive bands/zones were observed in 1D blot which resolved to 12 allergo-reactive spots in the 2D blot. Mass spectrometric analysis identified nine spots that grouped into four diverse proteins. Pathogenesis-related Thaumatin-like protein was found to be one of the major allergens in Lantana camara.
This is to our knowledge the first attempt to identify allergens from Lantana camara using a proteomic approach. The allergens identified thereof can be used to prepare hypoallergenic vaccine candidates and design immunotherapy trials against LC pollen and other aeroallergen carriers which are cross-reactive and harbor similar proteins.
过敏性疾病的发病率正在逐渐上升,是影响社会经济生活质量的全球性负担。过敏原的鉴定是为特应性疾病的治疗干预铺平道路的第一步。我们之前的调查发现,在印度的一个亚热带城市,总花粉负荷与呼吸道过敏的增加显著相关。在大多数患者中,导致IgE敏感性的最主要花粉似乎来自马缨丹(LC),这是一种生长在西孟加拉邦郊区及其周边的有害杂草。在本研究中,我们采用免疫蛋白质组学方法从这种灌木中鉴定出了过敏原成分。
连续两年通过空气生物学采样确定主要花粉种类,并将其与住院情况和皮肤点刺试验相关联。从LC阳性患者中采集血清,使用酶联免疫吸附测定(ELISA)和组胺测定法检测体外过敏原性。在十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS-PAGE)、二维聚丙烯酰胺凝胶电泳(2D PAGE)中分析总蛋白质组,并进行免疫印迹以检测IgE结合蛋白,然后使用质谱进一步鉴定这些蛋白。
通过与呼吸道过敏患者住院情况的相关性研究发现,马缨丹花粉是一个重要因素,其提取物在ELISA和组胺释放试验中显示出升高的IgE反应。在一维印迹中观察到5个IgE反应条带/区域,在二维印迹中解析为12个过敏反应斑点。质谱分析鉴定出9个斑点,这些斑点归为4种不同的蛋白质。病程相关类thaumatin蛋白被发现是马缨丹中的主要过敏原之一。
据我们所知,这是首次尝试使用蛋白质组学方法从马缨丹中鉴定过敏原。鉴定出的过敏原可用于制备低变应原性疫苗候选物,并设计针对LC花粉和其他具有交叉反应且含有相似蛋白质的气传过敏原载体的免疫治疗试验。