TLR2 ligands of chlamydiae

衣原体TLR2配体

基本信息

批准号：
7700302
负责人：
CATHERINE MARY O'CONNELL
金额：
$ 18.94万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-18 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7700302
关键词：
Antibiotic Therapy Antigens Bacteria Biochemical Blindness Bone Marrow Carbohydrates Cell Culture Techniques Cells Characteristics Chlamydia Chlamydia Infections Chlamydia trachomatis Cicatrix Classification Complex Data Dendritic Cells Development Disease Drug Delivery Systems Ectopic Pregnancy Eye Infections Genes Genital system Genome Glycogen Goals High Prevalence Immune response Immunologic Adjuvants Incidence Infection Infection prevention Infertility Inflammatory Ligands Lipoprotein (a)Lipoproteins Mammalian Oviducts Membrane Modification Molecular Molecular Profiling Molecular Weight Mus Parents Pathology Pathway interactions Pelvic Inflammatory Disease Plasmids Polymers Process Production Property Proteins Proteomics Publishing Randomized Recombinants Regulatory Element Reporting Risk Estimate Sexually Transmitted Diseases Signal Pathway Signal Transduction Tissues Toll-Like Receptor 2 Trachoma Two-Dimensional Gel Electrophoresis Vaccines Virulent Woman chronic pelvic pain computerized data processing cost cytokine immunopathology in vivo men mutant novel novel strategies public health relevance receptor reproductive reproductive development response tool trafficking two-dimensional

项目摘要

DESCRIPTION (provided by applicant): Chlamydia trachomatis infections are the leading cause of involuntary infertility and ectopic pregnancy in women. Although antibiotic treatment cures infection, it does not ameliorate the inflammatory process that leads to the development of disease. We have previously demonstrated that Toll-like receptor 2 (TLR2)-deficient mice fail to develop oviduct pathology after chlamydial infection, demonstrating that TLR2 signaling is directly involved in disease development. Identification of the bacterial product responsible for induction of TLR2-dependent signaling in response to chlamydial infection is essential to advance our ultimate goal of determining the molecular mechanisms that drive the development of chlamydia-induced immunopathology. We have derived novel, plasmid-deficient C. muridarum mutants that retain the ability to infect the murine genital tract but do not develop the characteristic upper tract scarring and pathology of chlamydial disease because they fail to stimulate TLR2-dependent signaling in cell culture and in vivo. We hypothesize that a gene product or regulatory element transcribed from the chlamydial plasmid influences expression or modification of pathogenic TLR2 ligand(s). In this application we propose careful, systematic proteomic and biochemical approaches to identify and characterize the chlamydial component that drives the development of upper tract disease. Specifically we will: (1) employ two-dimensional difference gel electrophoresis (2D-DIGE) to resolve differences in the expression profile of the plasmid-deficient strain when compared to its parent, and the use of a TLR2-Fc fusion construct as a way to identify candidate TLR2 ligands and to characterize ligand-receptor complexes biochemically; (2) examine the immunomodulatory properties of chlamydial glycogen which is no longer synthesized by plasmid-deficient chlamydiae and (3) determine the contribution of chlamydial lipoproteins to the induction of TLR2-dependent cytokine expression. Plasmid-deficient chlamydiae express Mip, a recently-identified candidate chlamydial TLR2 ligand, normally, suggesting that native chlamydial lipoproteins do not contribute to the development of reproductive tract disease. Identification of the pathogenic TLR2 ligand(s) will serve several purposes. We will be able to evaluate expression of the TLR2-dependent ligands by the plasmid-deficient strains. The genes or pathways involved in the expression of the ligand may serve as drug targets for therapies that limit tissue damage arising from infection. It may aid in the identification of endogenous TLR2 activating ligands that might contribute to the development of tubal scarring. Finally, it is possible that the TLR2 ligand of C. trachomatis could serve as an immunogen, eliciting protective responses that block or down-regulate TLR2-dependent signaling in response to chlamydial infection and provide a novel approach to protecting the upper reproductive tract from deleterious sequelae. PUBLIC HEALTH RELEVANCE: Chlamydia trachomatis infections are the leading cause of involuntary infertility and ectopic pregnancy in women. Tissue damage and scarring of the upper reproductive tract that occurs is the result of an immune response triggered by an as yet unknown molecule, or ligand, made by the bacteria. This application is to use novel mutants of C. trachomatis that cannot stimulate this signaling process as tools to identify this ligand and to examine the stimulatory properties of a carbohydrate polymer and a lipoprotein made by chlamydiae to determine if they also contribute to this damaging signal pathway.

描述（由申请人提供）：沙眼衣原体感染是女性非自愿不育症和异位妊娠的主要原因。尽管抗生素治疗可以治愈感染，但并不能改善导致疾病发展的炎症过程。我们以前已经证明，类似Toll样受体2（TLR2）缺陷小鼠在衣原体感染后未能发展出卵形病理学，这表明TLR2信号直接参与疾病发展。鉴定负责诱导TLR2依赖性信号转导对衣原体感染的细菌产品对于促进我们确定驱动衣原体诱导的免疫病理发展的分子机制的最终目标至关重要。我们衍生出了新颖的质粒缺陷型鼠梭状芽孢杆菌突变体，它们保留了感染鼠类生殖道的能力，但并未发展出特征性的上层疤痕和衣原体疾病的病理学，因为它们无法刺激细胞培养和体内的TLR2依赖性信号。我们假设从衣原体质粒转录的基因产物或调节元件会影响致病性TLR2配体的表达或修饰。在此应用中，我们提出了仔细的，系统的蛋白质组学和生化方法，以识别和表征驱动上层疾病发展的衣原体成分。具体而言，我们将：（1）使用二维差异凝胶电泳（2D-DIGE）来解决质粒缺乏菌株的表达谱的差异，与其父母相比，使用TLR2-FC融合构建体的使用作为候选TLR2配体和识别配体 - 培养基复合物Bioecteres Bioceptores biiochemecementes biochemchemchemsemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchemchem cossemate; （2）检查衣原体糖原的免疫调节特性不再由缺乏质粒的衣原体合成，（3）确定衣原体脂蛋白对TLR2依赖性细胞因子表达的诱导的贡献。质粒缺陷型衣原体Express MIP是最近被识别的候选衣原体TLR2配体，通常表明天然衣原体脂蛋白不影响生殖道疾病的发展。致病性TLR2配体（S）的鉴定将有几个目的。我们将能够通过质粒缺陷菌株评估TLR2依赖性配体的表达。参与配体表达的基因或途径可以作为限制感染引起的组织损伤的疗法的药物靶标。它可能有助于鉴定内源性TLR2激活可能有助于输卵管疤痕发展的配体。最后，沙眼梭状芽孢杆菌的TLR2配体可能充当免疫原，引起保护性反应，以响应衣原体感染的反应阻断或下调TLR2依赖性信号传导，并提供一种新的方法，可以保护上繁殖型的上层生殖型seeelae。公共卫生相关性：沙眼衣原体感染是女性非自愿不育症和异位妊娠的主要原因。发生的上部生殖道的组织损伤和疤痕是由细菌制造的尚未清楚的分子或配体触发的免疫反应的结果。该应用是使用不能刺激该信号传导过程的新型突变体作为识别该配体的工具，并检查碳水化合物聚合物的刺激特性和衣原体生成的脂蛋白，以确定它们是否也有助于这种有害的信号途径。