Pilot--Immunomodulation /Vaccine Optimization /Chlamydia

沙眼衣原体生殖器感染是最常见的细菌性传播疾病。 在美国，每年花费在400多万个临床病例上的费用超过20亿美元。就像几个 其他需要强大的辅助性T细胞1型(Th1)免疫来控制的细胞内细菌病原体(例如， 李斯特菌和分枝杆菌)，目前还没有针对沙眼衣原体的疫苗。人乳头瘤病毒免疫生物学研究进展 衣原体已经表明，识别和靶向免疫调节因子的新方法可以调节 Th1细胞的诱导对于设计针对这些病原体的有效疫苗至关重要。最近，它是 发现衣原体致敏、白介素10缺乏的树突状细胞(DC)具有很强的抗原提呈能力 诱导快速而强大的Th1反应和补充性体液免疫反应的细胞 赋予小鼠对生殖器衣原体感染的不育性免疫。衣原体冲洗剂的疗效观察 IL-10缺陷的DC不仅仅是由于缺乏IL-10，而是由于获得了某些 免疫生物学特性，包括快速成熟和表达一套独特的免疫调节因子 分子。本研究的主要目的是阐明黄斑狼疮的分子和免疫生物学基础。 衣原体致敏的IL-10缺陷树突状细胞的效力，包括定义可以 应用于设计和生产有效的衣原体疫苗。核心假设是 检测表明，衣原体致敏的IL-10缺陷树突状细胞在数量和质量上是不同的 与野生型(WT)DC相比，与T细胞激活相关的代谢特征。去调查 这一假设，我们将使用蛋白质组学和免疫学技术的组合，包括二维 凝胶电泳法(2-DE)、基质辅助激光解吸电离飞行时间法(MALDI-TOF)、 小干扰RNA(SiRNA)体内基因沉默及基因工程特异性基因分析 基因敲除或转基因小鼠，以识别和免疫学特征特定的基因产物 以促进DC的早期成熟和促进有效的APC功能，从而增强Th1的激活。额外的体内试验 体外生化技术将被用来在体内传递衣原体特异性疫苗 相关分子的存在或不存在，以确定对体内疫苗效力的影响，在 生殖器衣原体感染的小鼠模型。最终目标是识别和描述某些 免疫调节分子，可用于设计和提供有效的衣原体疫苗。

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.