MWF Mycobacteria antigens in occupational hypersensitivity pneumonitis

描述（由申请人提供）：机械工人的职业性过敏性肺炎（HP）与分枝杆菌有关，分枝杆菌经常定植在现代水基金属加工液（MWFs）中。HP是一种由吸入微生物抗原引起的免疫介导的肺部疾病，在其诊断、发病机制和治疗方面的研究相对较少。本研究的总体目标是鉴定和表征与MWF分枝杆菌种M. immunogenum （MI）和M. chelonae （MC）诱导HP潜能相关的抗原，并评估它们在HP免疫诊断和暴露评估中的潜力。在以前的资助周期中，我们已经分离出具有可变抗原特征的两种物种的多个菌株（基因型）。我们将验证我们的假设，即商业MWF中流行的MWF分枝杆菌菌株的抗原多样性是导致HP症状和潜在免疫反应差异的原因，并且负责的特异性抗原可以形成更有信息的诊断和个人和环境暴露评估的基础。具体目的是：(1)。在优化的HP小鼠模型中，基于MI和MC菌株与宿主免疫系统的相对相互作用，鉴定关键抗原；（2）. 通过克隆、蛋白表达和特异性抗体的产生重组制备鉴定出的关键抗原；(3)。探讨特异性抗原在有症状和无症状个体的早期个人暴露评估和免疫诊断中的潜力；(4)。评估MWF分枝杆菌的特异性抗原在环境暴露评估（流体监测）中的潜力。利用我们优化的小鼠模型，基于HP诱导电位和细胞介导的免疫反应，比较分离的MI(6)和MC(4)不同基因型的相对免疫原性，并通过差异免疫蛋白质组学分析鉴定推定的蛋白抗原。将利用小鼠模型和人类受试者评估重组表达抗原的免疫诊断潜力，特别是在区分无症状（暴露）与有症状（HP患者）以及环境暴露评估方面。由此产生的信息将有助于NORA的目标，为分枝杆菌暴露评估和HP免疫诊断提供一套工具；这些结果将有助于未来的流行病学研究和制定长期MWF暴露的预防（疫苗接种）和干预策略。

DESCRIPTION (provided by applicant): Occupational hypersensitivity pneumonitis (HP) in machine workers has been associated with mycobacteria that frequently colonize modern water-based metalworking fluids (MWFs). HP is an immune-mediated lung disease caused by inhalation of microbial antigens and is relatively poorly studied in terms of its diagnosis, pathogenesis, and treatment. Overall goal of this study is to identify and characterize antigens associated with HP-inducing potential of MWF mycobacteria species M. immunogenum (MI) and M. chelonae (MC) and evaluate their potential for HP immunodiagnosis and exposure assessment. In previous funding cycles, we have isolated multiple strains (genotypes) of both the species with variable antigenic characteristics. We will test our hypothesis that antigen diversity in MWF mycobacteria strains prevalent in commercial MWF is responsible for differentially inducing the HP symptoms and underlying immune response, and that the responsible specific antigens could form the basis for more informative diagnosis and personal and environmental exposure assessment. The specific aims are to: (1). Identify key antigens in MI and MC strains based on their relative interaction with the host immune system in an optimized mouse model of HP; (2). Recombinant production of the identified key antigens by cloning and protein expression and generation of specific antibodies; (3). Investigate potential of the specific antigens for early personal exposure assessment and immunodiagnosis in symptomatic versus asymptomatic individuals; (4). Evaluate potential of the specific antigen(s) of MWF mycobacteria for environmental exposure assessment (fluid monitoring). Relative immunogenicity of the isolated different genotypes of MI (6) and MC (4) will be compared based on HP induction potential and cell-mediated immune response using our optimized mouse model and the putative protein antigens will be identified by differential immunoproteomic profiling. The recombinantly expressed antigens will be evaluated for their potential for immunodiagnosis using the mouse model and human subjects, particularly in terms of differentiation of the asymptomatic (exposed) versus symptomatic (HP patients) and for environmental exposure assessment. The resulting information will help NORA's objectives by providing a set of tools for mycobacterial exposure assessment and HP immunodiagnosis; these outcomes will facilitate future epidemiological studies and development of prevention (vaccination) and intervention strategies for MWF exposures in long-term. PUBLIC HEALTH RELEVANCE: Non-tuberculous mycobacteria (NTM) colonizing modern metalworking fluids (MWFs) have been associated with occupational respiratory illnesses. Recent studies by NIOSH and other groups have linked MWF mycobacteria with occupational hypersensitivity pneumonitis (HP) in exposed machine workers. HP is an immune-mediated lung disease caused by inhalation of microbial antigens. Hence, characterization of specific antigens in MWF mycobacteria and evaluation of their potential for HP immunodiagnosis and exposure assessment will facilitate development of pre-emptive measures and intervention strategies in MWF-associated occupational environments.