Pathogenomics of Chlamydial Infection

衣原体感染的病理基因组学

• 批准号: 8156915
• 负责人: HARLAN D CALDWELL
• 金额: $ 67.49万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 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/8156915
• 关键词: Animal Model; Antibody Formation; Biology; CD8B1 gene; Characteristics; Chlamydia Infections; Cytotoxic T-Lymphocytes; Dendritic Cells; Development; Disease; Environment; Eukaryotic Cell; Female; Functional disorder; Future; Generations; Genes; Genital system; Goals; HIV; Human; Immune; Immune system; Immunity; Infection; Investigation; Laboratories; Malignant neoplasm of cervix uteri; Membrane Proteins; Molecular Conformation; Mus; Mutation; Pathogenesis; Pathology; Proteins; Recombinants; Risk Factors; Role; Sexually Transmitted Diseases; Structural Protein; Structure; T-Lymphocyte; Vaccine Design; Vaccines; Virulence; Virulence Factors; Virulent; Woman; Work; base; extracellular; immune function; in vivo; insight; neutralizing antibody; success; transmission process

Studies are focused on understanding the structure and function of the chlamydial polymorphic membrane protein D (PmpD) to better define its role in the biology of chlamydial infection and in its interaction with the host immune system. We have purified to homogeneity by immunoaffinity purification a native soluble fragment of PmpD that is secreted into the extracellular environment late in the infection cycle. The purified soluble fragment is being characterized at the protein level to define its secondary structural characteristics, and biologically by studying the interaction of the native protein with culture eukaryotic cells, human T cells, and dendritic cells. These studies are aimed at understanding the potential role of PmpD as virulence factor that might function in the suppression of chlamydial specific immune functions; specifically CD8 cytotoxic T cell immunity. PmpD is highly conserved antigenically among all C. trachomatis serovariants therefore a subunit PmpD vaccine based on the generation of a broadly cross-reactive protective antibody response is an attractive and ongoing goal of the laboratory. As the conformation of PmpD is essential for its ability to generate highly efficacious protective neutralizing antibodies we are using our protein structural analysis findings to design recombinant expression strategies capable of mimicking the proteins native structure. Although challenging, the success of these investigations are likely critical to the development of an efficious PmpD based vaccine. Using in vivo selection we isolated a hypervirulent C. trachomatis human urogential strain. The virulent isolate produces infection and disease in the mouse female genital tract with similar pathology to that of human infection. These findings are the first description of a human strain that is virulent for the mouse thereby providing a much needed small animal model for the study of human infection and disease. Remarkably, comparative genomic studies of virulent and avirulent clonal strains revealed a mutation in only a single gene unambiguously identifying the gene (CT153) as a critical in vivo virulence factor. These findings will enhance our understanding of the pathophysiology of human disease and better define at the cellular level immune mechanisms important to the devopment of protective immunity to infection. Future studies will focus on understanding how CT135 exacerbates infection and whether CT135 mutations are associated with chlamydial virulence in human disease. Defining the role of CT135 in pathogenesis could provide new insights important to chlamydial vaccine design and development.

研究的重点是了解衣原体多态性膜蛋白D（PmpD）的结构和功能，以更好地定义其在衣原体感染的生物学中的作用，以及其与宿主免疫系统的相互作用。 我们已经通过免疫亲和纯化将在感染周期后期分泌到细胞外环境中的PmpD的天然可溶性片段纯化至均一。 纯化的可溶性片段在蛋白质水平上进行表征，以确定其二级结构特征，并通过研究天然蛋白质与培养的真核细胞、人T细胞和树突细胞的相互作用进行生物学表征。 这些研究旨在了解PmpD作为毒力因子的潜在作用，其可能在抑制衣原体特异性免疫功能中起作用;特别是CD8细胞毒性T细胞免疫。 PmpD在所有C.因此，基于产生广泛交叉反应性保护性抗体应答的亚单位PmpD疫苗是实验室的一个有吸引力的和正在进行的目标。 由于PmpD的构象对于其产生高效保护性中和抗体的能力至关重要，我们正在使用我们的蛋白质结构分析结果来设计能够模拟蛋白质天然结构的重组表达策略。 虽然具有挑战性，但这些研究的成功可能对开发有效的基于PmpD的疫苗至关重要。 采用体内筛选的方法，我们分离到一株高毒力的C.沙眼人泌尿生殖道菌株 强毒株在小鼠雌性生殖道中产生感染和疾病，其病理学与人类感染的病理学相似。 这些发现首次描述了对小鼠具有毒性的人类菌株，从而为人类感染和疾病的研究提供了急需的小动物模型。 值得注意的是，毒性和无毒克隆菌株的比较基因组研究揭示了一个突变，只有一个基因明确地确定该基因（CT153）作为一个关键的体内毒力因子。 这些发现将增强我们对人类疾病的病理生理学的理解，并更好地在细胞水平上定义对感染保护性免疫的发展至关重要的免疫机制。未来的研究将集中在了解CT135如何加剧感染以及CT135突变是否与人类疾病中的衣原体毒力相关。 确定CT135在发病机制中的作用可以为衣原体疫苗的设计和开发提供重要的新见解。