Pilot--Immunomodulation and Vaccine Optimization against Chlamydia

中试——针对衣原体的免疫调节和疫苗优化

• 批准号: 7906787
• 负责人: QING HE
• 金额: $ 5.29万
• 依托单位: MOREHOUSE SCHOOL OF MEDICINE
• 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/7906787
• 关键词: Agonist; Antibodies; Antigen-Presenting Cells; Bacterial Sexually Transmitted Diseases; Biochemical; Cell Maturation; Characteristics; Chlamydia; Chlamydia Infections; Chlamydia trachomatis; Clinical; Clinical Research; Data; Dendritic Cells; Development; Ectopic Pregnancy; Effectiveness; Elements; Engineering; Gene Silencing; Genes; Genital system; Genus Mycobacterium; Goals; Health Care Costs; Immune; Immune response; Immunity; Immunobiology; Immunologic Techniques; Immunologics; In Vitro; Infection; Infertility; Interleukin-10; Interleukins; Knock-out; Knockout Mice; Lead; Liquid Chromatography; Listeria; Metabolic; Methods; Modeling; Molecular; Mus; Pelvic Inflammatory Disease; Physiologic pulse; Property; Proteins; Proteome; Proteomics; RNA; RNA analysis; Role; Sexually Transmitted Diseases; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; T-Cell Activation; T-Lymphocyte; Techniques; Technology; Testing; Th1 Cells; Time; Transgenic Mice; Transgenic Organisms; Two-Dimensional Gel Electrophoresis; United States; Vaccines; Woman; base; design; gel electrophoresis; genital infection; immunoregulation; in vivo; knockout gene; medical schools; microbial; novel; novel strategies; pathogen; programs; protein profiling; response; stimulant/agonist; tandem mass spectrometry; vaccine efficacy

Genital infection by Chlamydia trachomatis is the most common bacterial sexually transmitted disease (STD) in the United States with an excess of $2 billion spent on over four million annual clinical cases. Like several other intracellular bacterial pathogens that require a robust T helper type 1 (Th1) immunity for control (e.g., Listeria and Mycobacteria), there are no vaccines against C. trachomatis. Progress in the immunobiology of Chlamydia has indicated that novel approaches to identify and target immunomodulatory factors that regulate the induction of Th1 cells are crucial for designing effective vaccines against these pathogens. Recently, it was found that chlamydia-pulsed, interleukin (IL)-10 deficient dendritic cells (DCs) were potent antigen-presenting cells that induced a rapid and robust Th1 response and the complementary humoral immune response which conferred sterilizing immunity against genital chlamydial infection in mice. The effectiveness of chlamydiapulsed IL-10 deficient DCs is not due merely to the absence of IL-10 but to acquisition of certain immunobiologic properties that include rapid maturation and expression of a unique set of immunomodulatory molecules. The main objective of this study is to elucidate the molecular and immunobiological basis for the potency of chlamydia-pulsed IL-10 deficient DCs, including defining novel molecular elements that can be applied in designing and delivering efficacious vaccines against Chlamydia. The central hypothesis to be tested is that chlamydia-primed IL-10 deficient DCs are quantitatively and qualitatively distinct in their metabolic characteristics relating to T cell activation compared to wild-type (WT) DCs. To investigate this hypothesis, we will use a combination of proteomics and immunological techniques, including twodimensional gel electrophoresis (2-DE), matrix-assisted laser-desorption/ionization time-of-flight (MALDI-TOF), in vivo gene silencing via short interfering RNA (siRNA), and analysis of genetically-engineered specific gene knockout or transgenic mice, to identify and immunologically characterize certain gene products that contribute to early DC maturation and promote efficient APCs function for an enhanced Th1 activation. Additional in vivo and ex vivo biochemical techniques will be used to deliver chlamydia-specific vaccine constructs in the presence or absence of the relevant molecules identified, to determine the effect on vaccine efficacy in vivo, in a murine model of genital Chlamydia infection. The ultimate goal is to identify and characterize certain immunomodulatory molecules that can be used to design and deliver efficacious vaccines against Chlamydia.

生殖道沙眼衣原体感染是最常见的细菌性传播疾病（STD） 在美国，每年花费超过20亿美元用于超过400万例临床病例。像几 其他需要强大的1型辅助性T细胞（Th 1）免疫力来控制的细胞内细菌病原体（例如， 李斯特菌和分枝杆菌），目前还没有针对C。沙眼免疫生物学研究进展 衣原体已经表明，新的方法来确定和靶向免疫调节因子， Th 1细胞的诱导对于设计针对这些病原体的有效疫苗是至关重要的。近日有 发现衣原体冲击的白细胞介素（IL）-10缺陷树突状细胞（DC）是有效的抗原呈递细胞， 诱导快速和强大的Th 1应答和互补的体液免疫应答的细胞， 在小鼠中赋予针对生殖器衣原体感染的绝育免疫。衣原体感染的有效性 IL-10缺陷型DC不仅仅是由于缺乏IL-10，而是由于获得了某些IL-10受体。 免疫生物学特性，包括快速成熟和表达一组独特的免疫调节因子， 分子。本研究的主要目的是阐明这种疾病的分子和免疫生物学基础， 衣原体脉冲的IL-10缺陷型DC的效力，包括定义可以 用于设计和提供有效的衣原体疫苗。核心假设是 测试的是衣原体致敏的IL-10缺陷型DC在它们的免疫应答中定量和定性不同。 与野生型（WT）DC相比，与T细胞活化相关的代谢特征。探讨 这个假设，我们将使用蛋白质组学和免疫学技术的组合，包括二维 凝胶电泳（2-DE），基质辅助激光解吸/电离飞行时间（MALDI-TOF）， 通过短干扰RNA（siRNA）体内基因沉默，以及基因工程特异性基因的分析 基因敲除或转基因小鼠，以鉴定和免疫学表征某些基因产物， 早期DC成熟和促进有效的APC功能，以增强Th 1活化。额外的体内 和离体生物化学技术将用于在体内递送衣原体特异性疫苗构建体 鉴定的相关分子的存在或不存在，以确定对体内疫苗效力的影响， 生殖器衣原体感染的小鼠模型。最终目标是识别和表征某些 免疫调节分子，可用于设计和提供针对衣原体的有效疫苗。