NOVEL CELLULAR & GENETIC ANALYSES USING A NEW CHLAMYDIA PENETRANT PEPTIDE

新颖的蜂窝

基本信息

批准号：
8281426
负责人：
Ashok A Aiyar
金额：
$ 17.75万
依托单位：
LSU HEALTH SCIENCES CENTER
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-06-15 至 2014-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8281426
关键词：
Address Adverse effects Animals Antisense Oligonucleotides Attenuated Bacterial Genes Biology Case Study Cells Chlamydia Chlamydia trachomatis Chronic Chronic Disease Clinical Data Columnar Epithelium Cytokine Inducible SH2-Containing Protein Data Development Dioxygenases Disease Education Endocervix Epithelial Cells Female Fertility Genes Genetic Genetic Techniques Genital system Genitourinary system Goals Habitats Health Health Care Costs Hour Human Immune Immune Targeting Immune response Immunology In Vitro Infection Interferon Type II Interferons Intervention Invaded Label Life Mediating Mediator of activation protein Methods Molecular Molecular Biology Neonatal Nucleic Acids Organism Pathogenesis Pathway interactions Peptide Nucleic Acids Peptides Physiological Population Protein Biosynthesis Proteins Public Health Regulation Research Role Screening procedure Signal Transduction Sorting - Cell Movement Technology Testing Time Tryptophan United States National Institutes of Health Virulence Woman attenuation genetic analysis human female in vivo indoleamine innovation men neonate novel pathogen positional cloning reproductive research study response skills tool transmission process

项目摘要

DESCRIPTION (provided by applicant): The objectives of this proposal are to elucidate the mechanism by which Chlamydia trachomatis evades the immune response by attenuating IFN?-dependent signaling in infected host cells, and to develop a new reverse genetic strategy to study C. trachomatis pathogenesis and biology. Sexually transmitted C. trachomatis infections, primarily caused by serovars D-F, are a major public health concern with adverse effects on female fertility and neonatal health. Despite significant public health interventions including education, screening and treatment, reported cases n the US reached 1 million in 2006 and continue to rise, resulting in annual health care costs >$2.4 billion. Asymptomatic infections in >70% of women and >30% of men, can take months to years to resolve, and thereby compounding transmission of C. trachomatis. Transmission is exacerbated by short-lived protection to re-infection. In vitro studies in human epithelial cells, and animal studies using C. muridarum, have demarcated IFN? as a major anti-chlamydial mediator. In vivo, although elevated IFN??is found in the infected human endocervix, clinical data indicate that C. trachomatis can evade immune responses. We have developed an innovative approach using a novel penetrant peptide (CPP) to label C. trachomatis Elementary Bodies (EB). Our approach has permitted the isolation of pure populations of infected endocervical epithelial cells, from mixed pools of cells exposed to C. trachomatis. Analyses of pure populations has revealed that C. trachomatis attenuates IFN?-dependent expression of indoleamine-2,3-dioxygenase (IDO1). Attenuation permits C. trachomatis, a tryptophan auxotroph, to ameliorate the effect of intracellular tryptophan depletion by IDO1. The first goal of this application is to understand how Chlamydia blocks host-cell IFN?-dependent signaling. Understanding this mechanism will permit development of new strategies that target immune evasion by C. trachomatis. The paucity of easily applicable genetic tools to manipulate C. trachomatis has limited the capacity to investigate mechanisms employed by C. trachomatis that drive immune evasion and pathogenesis. This application addresses this significant deficiency by proposing an innovative approach using the same CPP to carry cargoes that repress expression of specific C. trachomatis genes. When developed and validated, our strategy will permit robust reverse genetic approaches to molecularly dissect virulence determinants of C. trachomatis. These studies with our novel CPP will significantly shift current paradigms and revolutionize approaches in Chlamydia research by: 1) Establishing a robust method to elucidate previously unidentified immune evasion strategies used by C. trachomatis; and 2) Providing, for the first time, a robust reverse genetic technique to dissect the role of specific bacterial genes in the developmental cycle and pathogenesis.

描述（由申请人提供）：该提案的目的是阐明沙眼衣原体通过减弱感染宿主细胞中IFN依赖性信号来逃避免疫反应的机制，并开发了一种研究新的反向遗传策略来研究C. c. c. c. c. c. trolachomatis病原体和生物学。性传播的梭菌感染主要由血清D-F引起，是对女性生育能力和新生儿健康的不利影响的主要关注。尽管公共卫生的干预措施包括教育，筛查和治疗，但报告的病例在2006年达到100万，并继续增加，导致年度医疗保健费用> 24亿美元。 > 70％的女性和> 30％的男性的无症状感染可能需要数月到几年才能解决，从而使沙眼梭状芽胞盘的传播更加复杂。短暂的保护以重新感染加剧了传播。在人类上皮细胞和使用Muridarum C.的动物研究中进行体外研究已划分IFN？作为主要的抗神经介质。在体内，尽管在被感染的人内核中发现了IFN的升高，但临床数据表明沙眼梭状芽孢杆菌可以逃避免疫反应。我们已经使用新型的渗透肽（CPP）开发了一种创新的方法，以标记沙眼基本体（EB）。我们的方法允许从暴露于沙眼藻的混合细胞中分离出感染的遗传性宫颈上皮细胞的纯种群。对纯种群的分析表明，沙眼梭状芽孢杆菌可减轻IFN？依赖性吲哚胺-2,3-二氧酶的表达（IDO1）。衰减允许色氨酸合理营养杆菌C. c. c. c. themoprophroph，可以通过IDO1改善细胞内色氨酸耗竭的作用。该应用的第一个目标是了解衣原体如何阻止宿主网络IFN？依赖性信号传导。了解这种机制将允许制定针对沙眼甲状骨炎挑战的新策略。易于适用的遗传工具操纵沙丘梭状芽孢杆菌的缺乏限制了调查驱动免疫逃避和发病机理的沙眼使用的机制的能力。该应用程序通过提出使用相同CPP的创新方法来解决这种重大缺陷，以携带抑制特定沙眼基因表达的货物。当开发和验证时，我们的策略将允许鲁棒的反向遗传方法分子剖析沙眼曲霉的毒力决定因素。这些对我们的新型CPP的研究将显着改变当前的范式，并通过以下方式彻底改变衣原体研究中的方法：1）建立一种可靠的方法来阐明先前未鉴定的弯曲孢霉菌的免疫逃避策略； 2）首次提供一种强大的反向遗传技术，可以剖析特定细菌基因在发育周期和发病机理中的作用。