Molecular biological study of the pathogenesis of summer-type hypersensitivity pneumonitis

根据分子数据，我们确定夏季型超敏性肺炎（SHP）的病原体为阿萨希毛孢霉和粘液样衣原体。在本研究中，我们应用聚合酶链反应（PCR）和Southern杂交技术检测了小鼠模型或SHP患者支气管肺泡灌洗液（BALF）和患者家中空气中的抗原特异性dna。吸入激射后6h和96h，模型BALF样品阳性率分别为100%和60%。18例患者入院后1周内采集BALF样本，94%为阳性。其他疾病患者23例，健康志愿者7例，分别为22%和29%。空气样本采用安徒生式小体积空气采样器采集。取样板上的颗粒用超声波清洗机采集，用PCR扩增。2/2的空气样本呈阳性。通过附加限制性内切酶Sau3AI的PCR，将ashii和T.mucoides的每个信号与其他真菌或人细胞区分开来。应用PCR技术检测宿主和环境中病原特异性dna，将为探讨微生物暴露职业病的发病机制提供一种可拓展的方法。

We assigned the causative agents of summer-type hypersensitivity pneumonitis(SHP) as Trichosporon asahii and T.mucoides, using molecular data. In this study, we applied polymerase chain reaction(PCR) and Southern hybridization to detect the antigen specific DNAs in the bronchoalveolar lavage fluid(BALF) of a murine model or patients with SHP, and in the air of patients' homes. BALF samples of the model were positive in 100% and 60% at 6h and 96h after the inhalation challenge, respectively. The BALF samples of 18 patients obtained within a week after admission were positive in 94%. Those of 23 patients with other diseases and 7 healthy volunteers were 22% and 29%, respectively. Air samples were collected by Andersen type low volume air sampler. Particles on the sampling plates were harvested by ultrasonic cleaner and amplified by PCR.The 2/2 air samples were positive. The each signal of T.asahii and T.mucoides were discriminated from the other fungi or human cells by PCR itself of additional restriction enzyme, Sau3AI.The detection of pathogen specific DNAs in host and environment by PCR would be an expandable method to investigate the pathogenesis of occupational diseases caused by microbial exposures.