Identification and characterization of fungal exposures

真菌暴露的鉴定和表征

• 批准号: 9430270
• 负责人: Donald Beezhold
• 金额: $ 8.13万
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9430270
• 关键词: Air; Allergic; Alternaria; Antibodies; Antigens; Area Analyses; Ascomycota; Asthma; Basidiomycota; Behavior Therapy; Biological Markers; Botrytis; Cannabis; Child; Data; Data Analyses; Development; Dideoxy Chain Termination DNA Sequencing; Dust; Environment; Environmental Risk Factor; Epitope Mapping; Evaluation; Exhalation; Exposure to; Extrinsic allergic alveolitis; Family; Geographic Locations; Goals; Gray unit of radiation dose; Health; Health Hazards; High-Throughput Nucleotide Sequencing; Home environment; Homologous Gene; House Dust; Housing; Hypersensitivity; Immunoassay; Income; Individual; Industry; Measures; Methodology; Methods; Molds; Molecular; Names; Neighborhoods; Nevada; New York City; Nitric Oxide; Occupational; Opportunistic Infections; Pathogenicity; Penicillium; Philadelphia; Plants; Prevalence; Production; Reaction; Recombinants; Ribosomal RNA; Sampling; Schools; Surveillance Methods; Systemic infection; Testing; United States; Vermont; Work; Xenobiotics; air sampling; airway inflammation; anthropogenesis; built environment; cross reactivity; design; experimental study; fungus; improved; insight; microbial; pathogen; polyclonal antibody; pullulan; rRNA Genes; transcriptome sequencing

To more completely assess the fungal species to which people are exposed, a fungal ribosomal RNA (rRNA) gene sequencing study was designed to test the hypothesis that fungal bioaerosols in the United States indoor built environments are much more diverse than previously estimated using traditional methods of analysis. Fungal rRNA sequencing data from samples collected from a variety of geographic regions within the United States including Atlanta (Georgia), Bennington (Vermont), Reno (Nevada) and Philadelphia (Pennsylvania) were analyzed using Illumina miSeq high throughput sequencing or Sanger sequencing to characterize fungal diversity in air and dust samples. In FY17 the project was extended to include the analysis of fungal diversity in occupational environmental samples collected in NIOSH Health Hazard Evaluations (HHEs). Analysis of area and personal air samples collected in the cannabis production industry has demonstrated a diversity of fungi primarily composed of species placed in the phylum Ascomycota including plant pathogenic species such as Botrytis cinerea, which causes grey mold. Other plant pathogens placed in the phylum Basidiomycota, including Wallemia sebi have also been enumerated in another recently sampled production facility. These HHEs have highlighted that workers can also be exposed to microbial contaminants including species that have been previously associated with occupational cases of hypersensitivity pneumonitis and allergic sensitization. The results of ITS rRNA gene sequencing studies have also provided new insights into the spectrum of fungi in occupational environments previously overlooked using traditional methods of assessment. Additional studies are examining the molecular diversity of fungal contaminants as part of the NYC Neighborhood Asthma and Allergy Study. Preliminary analysis of analyzed dust samples indicates that Aureobasidium pullulans, Penicillium glabrum, Wallemia sebi and Alternaria alternata varied by housing type (single, multi-family or apartment) and neighborhood asthma prevalence. Preliminary results suggest that multiple environmental factors including anthropogenic behavior modification, housing type, and neighborhood are important variables that influence fungal diversity within middle-income homes in New York City. A. alternata measured in house dust was also associated with fractional exhaled nitric oxide, specifically among children with higher combustion byproduct exposure, suggesting a possible interaction between these two exposures on airway inflammation. Fungal diversity captured using Illumina miSeq is being compared to results obtained using qPCR and the data analysis is ongoing. Work has continued in the development of antibodies to recombinant fungal biomarker antigens. The utility of these antibodies is critical for the quantification of fungal biomarkers, particularly to those fungi that are being studied by the NTP. NIOSH has recently developed a recombinant Alt a 1 homologue from Ulocladium chartarum tentatively named Ulo c 1. Polyclonal antibodies (pAb) have been produced toward the recombinant Ulo c 1. In FY17, pAb reactivity with rUlo c 1 as well as cross-reactivity with other Alt a 1 homologs has been determined and sandwich and inhibition ELISAs developed. Epitope mapping experiments and optimization of the immunoassay will continue in FY18.

为了更全面地评估人们接触的真菌物种，设计了一项真菌核糖体RNA（rRNA）基因测序研究，以测试美国室内建筑环境中的真菌生物气溶胶比以前使用传统分析方法估计的要多样化得多的假设。使用Illumina miSeq高通量测序或桑格测序来分析来自从美国内的各种地理区域收集的样品的真菌rRNA测序数据，所述地理区域包括亚特兰大（格鲁吉亚）、本宁顿（佛蒙特）、里诺（内华达州）和费城（宾夕法尼亚），以表征空气和灰尘样品中的真菌多样性。在2017财年，该项目扩展到包括NIOSH健康危害评估（HHE）中收集的职业环境样本中的真菌多样性分析。对在大麻生产行业收集的地区和个人空气样本的分析表明，真菌的多样性主要由子囊菌门的物种组成，包括植物病原物种，如灰葡萄孢，它会导致灰霉病。在另一个最近取样的生产设施中，也列举了属于担子菌门的其他植物病原体，包括沃尔菌。这些HHE强调，工人也可能暴露于微生物污染物，包括以前与职业性过敏性肺炎和过敏性致敏相关的物种。ITS rRNA基因测序研究的结果也为以前使用传统评估方法忽视的职业环境中的真菌谱提供了新的见解。其他研究正在检查真菌污染物的分子多样性，作为纽约市社区哮喘和过敏研究的一部分。初步分析分析的灰尘样品表明，出芽短梗霉，光滑青霉，瓦勒米sebi和链格孢菌不同的住房类型（单，多家庭或公寓）和邻里哮喘患病率。初步结果表明，多种环境因素，包括人为行为的修改，住房类型和邻里是重要的变量，影响真菌多样性的中等收入家庭在纽约市。A.在室内灰尘中测得的alternata也与呼出的一氧化氮分数相关，特别是在燃烧副产物暴露较高的儿童中，这表明这两种暴露对气道炎症的可能相互作用。使用Illumina miSeq捕获的真菌多样性正在与使用qPCR获得的结果进行比较，数据分析正在进行中。 针对重组真菌生物标志物抗原的抗体的开发工作仍在继续。 这些抗体的效用对于真菌生物标志物的定量是至关重要的，特别是对于NTP正在研究的那些真菌。NIOSH最近开发了一种来自Ulocladium chartarum的重组Alt a 1同源物，暂命名为Ulo c 1。针对重组Ulo c 1的多克隆抗体（pAb）已经产生。在2017财年，已经确定了pAb与rUlo c 1的反应性以及与其他Alt a 1同源物的交叉反应性，并开发了夹心和抑制ELISA。表位作图实验和免疫分析优化将在2018财年继续进行。