Department of Environmental Health Sciences, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, United States.
J Immunol Methods. 2013 Feb 28;388(1-2):86-9. doi: 10.1016/j.jim.2012.11.009. Epub 2012 Nov 29.
Fungal cell wall components, such as (1-3)-β-D-glucan, are known to be capable of activating the innate immune system and pose a respiratory health risk in different environments. Mass-based non-viable techniques commonly used for assessment of fungal exposures could be β-D-glucan-specific, but are limited to analysis of liquid extracts. The variable solubility of different β-D-glucans may underestimate β-D-glucan exposure and long sampling times required for mass-based methods make assessing short-term exposures difficult. In this study, we evaluated the utility of the halogen immunoassay (HIA), an immunoblotting technique previously used for allergens, to immunodetect and quantify β-D-glucan-carrying particles (BGCPs). The HIA was able to detect BGCPs without background staining when β-D-glucan standards and air samples collected at a poultry house during short sampling periods were evaluated. The image analysis protocol previously developed by our group for mouse allergen allowed simultaneous immunodetection and quantification of β-D-glucan-containing particles. Our results suggest that the HIA holds promise for quantifying β-D-glucan exposures. To our knowledge, this is the first time in which the HIA was used for non-allergenic compounds of microbial or fungal origins.
真菌细胞壁成分,如(1-3)-β-D-葡聚糖,已知能够激活先天免疫系统,并在不同环境中对呼吸道健康构成风险。常用于评估真菌暴露的基于质量的非存活技术可能是β-D-葡聚糖特异性的,但仅限于对液体提取物的分析。不同β-D-葡聚糖的可变溶解度可能会低估β-D-葡聚糖的暴露,而基于质量的方法所需的长时间采样使得评估短期暴露变得困难。在这项研究中,我们评估了卤化物免疫测定(HIA)的效用,该技术是一种先前用于过敏原的免疫印迹技术,用于免疫检测和定量携带β-D-葡聚糖的颗粒(BGCPs)。当评估β-D-葡聚糖标准品和在短时间采样期间在家禽养殖场收集的空气样本时,HIA 能够在没有背景染色的情况下检测到 BGCPs。我们小组之前为小鼠过敏原开发的图像分析方案允许同时免疫检测和定量含有β-D-葡聚糖的颗粒。我们的结果表明,HIA 有望定量β-D-葡聚糖暴露。据我们所知,这是 HIA 首次用于微生物或真菌来源的非过敏原化合物。