Core 1: Osteoimmunology Core

摘要 骨免疫学领域成立于2005年，旨在了解骨骼如何影响免疫， 造血细胞，以及造血和免疫细胞如何影响骨，在临床条件下， 人工关节感染（PJI）。然而，关键的工具和研究协议的发现，和技术 需要将发现转化为骨髓炎的临床诊断和干预措施，但当时并不存在。 我们的AO创伤临床优先计划（AO-CPP）于5年前开始。于是，我就把它收藏起来。 多学科研究团队在骨病理学、微生物发病机制 免疫学、电子显微镜、材料科学与工程、传染病和矫形外科， 使作为这个翻译研究中心的科学前提的重大进展能够在 骨感染的骨免疫学（CoRTOBI）。尽管有这些重大进展，额外的测定和 需要技术开发来完成该CoRTOBI中的研究项目。因此， 骨免疫学研究核心是：1）为CoRTOBI研究者提供最先进的研究 其他地方不存在的资源; 2）开发新的技术和分析方法， 骨免疫学研究。我们实现这些目标的方法体现在以下具体措施中： 目标。在目标1中，我们将处理样本，并对受感染的骨进行所有电子显微镜检查 并在项目1中制造纳米多孔膜;而“特洛伊木马”巨噬细胞和组织 在项目2中获得成功。在目标2中，我们将产生孔径不同的纳米多孔膜阵列， 评价S.金黄色葡萄球菌，并克隆基因 使用候选基因和TnSeq突变体文库筛选方法将其作为新的抗生素靶标。在 目标3：我们将在项目1中使用我们的3D打印技术来评估体内的新型抗生素间隔物。目标4 我们将筛选所有的血清从动物研究中，和300 PJI患者在AO-CPP注册，使用我们的 14-抗原多重Luminex检测，随着技术的发展，我们将增加额外的S.金黄色 我们的多重Luminex检测的抗原。在项目2中，我们还将开发新型的多重Luminex检测试剂盒， 对于鼠伤寒沙门氏epidermidis和S. lugdunensis，并使用它们来筛选小鼠和PJI患者的血清，正如我们所做的那样， 为S。金黄色。最后，在目标5中，我们将建立筛选和克隆分泌抗体的 细胞（ASC）的全血从小鼠和人类感染的S。金黄色。我们还将开发可复制的 方法扩增和克隆抗S.直接从小鼠和人血液中提取的金黄色葡萄球菌ASC作为一种新技术， 核心的重要的是，我们还打算与该CoRTOBI之外的研究者分享我们的资源， 并将我们的研究平台转移到其他研究葡萄球菌感染的机构， 和骨髓炎。 !

Abstract The field of Osteoimmunology was established in 2005 to understand how bone influences immune and hematopoietic cells, as well as how hematopoietic and immune cells influence bone, in clinical conditions like prosthetic joint infection (PJI). However, critical tools and research protocols for discovery, and technologies required to translate discoveries into clinical diagnostics and interventions for osteomyelitis, did not exist at the commencement of our AO Trauma Clinical Priority Program (AO-CPP) 5 years ago. Thus, we assembled a multidisciplinary team of investigators with extensive experience in bone pathology, microbial pathogenesis, immunology, electron microscopy, material science & engineering, infectious disease and orthopaedic surgery, to enable the major advances that serve as the scientific premise of this Center of Research Translation on the Osteoimmunology of Bone Infection (CoRTOBI). Despite these major advances, additional assay and technology development is needed to complete the research projects in this CoRTOBI. Thus, the goals of the Osteoimmunology Research Core are to: 1) provide the CoRTOBI investigators with state-of-the-art research resources that do not exist elsewhere; and 2) develop novel technologies and assays for future osteoimmunology research. Our approaches to achieve these goals are embodied by the following Specific Aims. In Aim 1 we will process the samples and perform all of the electron microscopy on the infected bone and fabricated nanoporous membranes in Project 1; and the “Trojan horse” macrophages and tissue abscesses in Project 2. In Aim 2 we will generate the array of nanoporous membranes varying in pore sizes to assess the haptotaxis, metamorphic and durotaxis behaviors of S. aureus in vitro, and clone the genes involved as novel antibiotic targets using a candidate gene and TnSeq mutant library screening approach. In Aim 3 we will use our 3D-printing technology to assess novel antibiotic spacers in vivo in Project 1. In Aim 4 we will screen all the sera from the animal studies, and the 300 PJI patients in the AO-CPP registry, using our 14-antigen multiplex Luminex assay, and as technology development, we will add additional S. aureus antigens to our multiplex Luminex assay. In Project 2, we will also generate novel multiplex-Luminex assays for S. epidermidis and S. lugdunensis, and use them to screen sera from mice and PJI patients, as we have done for S. aureus. Finally, in Aim 5 we will establish protocols for screening and cloning antibody-secreting cells (ASC) in whole blood from mice and humans infected with S. aureus. We will also develop reproducible methods to expand and clone anti-S. aureus ASC directly from murine and human blood as new technology in the Core. Importantly, it is also our intent to share our resources with investigators outside of this CoRTOBI, and transport our research platforms to other institutions with a research focus on Staphylococcal infections, and osteomyelitis. !