Biochemical, molecular and immunological characterization of Mycoplasma pneumoni

肺炎支原体是一种常见的非典型细菌病原体，与急性和慢性肺炎密切相关。 慢性呼吸道疾病，包括新发哮喘和病情恶化，以及#年持续感染 无论是孩子还是成年人。对肺炎支原体引起感染伴发炎性疾病的认识 通路、病理和后遗症一直是欠缺的。然而，我们对社区的认同 获得性呼吸窘迫综合征毒素(CADS TX)，一种真正的ADP核糖化和空泡化 毒素，在肺炎支原体和呼吸道炎症和损伤之间提供了直接的因果联系。这 肺炎支原体、卡介苗与肺部高反应性和炎症之间的联系 由SA-AADCRC研究工作加强，展示肺炎支原体和卡介苗TX直接临床 相关并得到动物建模和创新诊断、病理学、生物学、 化学和免疫学评估。考虑到这些发现以及结构和功能 百日咳和白喉毒素与卡介苗毒素的相似性和潜在致病意义 假设卡介苗Tx是肺炎支原体介导性哮喘的关键致病因子 与呼吸道相关的疾病。为了验证这一假设，我们将：1.表征TX介导的腺苷二磷酸核糖 转移酶(ART)活性通过检测ART功能所必需的卡片TX最小结构域并通过 鉴定被CADS TX ADP核糖化的哺乳动物靶蛋白。2.描述 通过识别最小区域(S)和介导TX结合的必需氨基酸来识别TX结合区 与表面活性蛋白-A和哺乳动物细胞受体的结合(S)。3.生成单克隆和多克隆 抗卡介苗Tx抗体的反应性及其表位的确定 阻断/中和抗体。在后一种情况下，抗体试剂的可用性可以中和和 阻止卡TX活动，以及在整个SA-AADCRC中概述的协作研究 应用应有助于治疗剂的创新设计和改进诊断。

Mycoplasma pneumoniae is a common atypical bacterial pathogen strongly associated with acute and chronic airway diseases, including new-onset asthma and exacerbations, as well as persistent infections in children and adults. Knowledge on how M. pneumoniae causes infection with accompanying inflammatory pathways, pathologies and sequelae has been deficient. However, our identification of the Community Acquired Respiratory Distress Syndrome ToXin (CARDS TX), a bona fide ADP ribosylating and vacuolating toxin, has provided a direct causal link between M. pneumoniae and ainway inflammation and injury. This connection between M. pneumoniae, CARDS TX and lung hyper-responsiveness and inflammation is reinforced by SA-AADCRC research efforts demonstrating M. pneumoniae and CARDS TX direct clinical relevance and further supported by animal modeling and innovative diagnostic, pathological, biological, chemical and immunological assessments. Considering these findings and the structural and functional similarities and potential pathogenic significance of CARDS TX relative to pertussis and diphtheria toxins, we hypothesize that CARDS TX is a critical pathogenic contributor to M. pneumoniae-mediated asthma and related ainway diseases. To test this hypothesis, we will: 1. Characterize CARDS TX-mediated ADP-ribosyl transferase (ART) activity by detecting the CARDS TX minimal domain essential for ART function and by identifying the mammalian target proteins that are ADP-ribosylated by CARDS TX. 2. Characterize the CARDS TX binding region by identifying the minimal region(s) and essential amino acids that mediate binding to surfactant protein-A and mammalian cell receptor(s). 3. Generate monoclonal and polyclonal antibodies reactive against CARDS TX and determine epitopes of CARDS TX capable of inducing blocking/neutralizing antibodies. In the latter case, the availability of antibody reagents that neutralize and block CARDS TX activities, along with the collaborative studies outlined throughout this SA-AADCRC application should assist in the innovative design of therapeutic agents and improved diagnostics.