Project 2: Defining the Protective vs. Susceptible Immune Proteome of S. aureus Osteomyelitis

项目 2：定义金黄色葡萄球菌骨髓炎的保护性与易感性免疫蛋白质组

• 批准号: 10247796
• 负责人: Edward M. Schwarz
• 金额: $ 32.45万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-20 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10247796
• 关键词: Animal Model; Antibodies; Antigens; Autoimmunity; Autolysin; Bacteria; Binding; Biological Assay; Blood; Cessation of life; Clinical; Complex; Custom; Data; Diabetes Mellitus; Endocytosis; Equus caballus; Event; Fc domain; Fibrinogen; Genus staphylococcus; Goals; Haptoglobins; Hemoglobin; Human; Humoral Immunities; Immune; Immune response; Immunologic Tests; Implant; In Vitro; Incidence; Infection; Intervention; Intracellular Transport; Iron; Joint Prosthesis; Kidney; Knowledge; Label; Leukocytes; Liver; LysoTracker; Mediating; Modeling; Molecular; Monoclonal Antibodies; Multiple Organ Failure; Mus; Obesity; Operative Surgical Procedures; Organ; Osteomyelitis; Outcome; Passive Immunization; Patients; Pilot Projects; Predisposition; Proteins; Proteome; Registries; Replacement Arthroplasty; Sepsis; Site; Spleen; Staphylococcal Protein A; Staphylococcus aureus; Staphylococcus aureus infection; Staphylococcus epidermidis; Stream; Surgical Wound Infection; Technology; Testing; Therapeutic immunosuppression; Time; Tissue Banks; Translating; Translational Research; Trauma; bactericide; fluorescence imaging; in vivo; indexing; joint infection; macrophage; man; osteoimmunology; programs; respiratory; septic; theories

Abstract Prosthetic joint infection (PJI) is the bane of elective total joint replacement (TJR) surgery, of which the vast majority is caused by Staphylococcal species. The 1-5% incidence of PJI is known to be a nonrandom event that is largely determined by patient specific factors. Moreover, ~13% of patients infected with S. aureus become septic and die from multiorgan failure, while others recover with little intervention. In addition to the established host susceptibility factors, there are two leading theories to explain this. The first is the “immune proteome” hypothesis, which posits that the array of specific antibodies a host develops against S. aureus dictates its susceptibility vs. protection to infection. The second is the “Trojan horse” model of S. aureus dissemination, in which leukocytes get infected at the surgical site, and transport intracellular bacteria to the blood stream and internal organs, culminating in septic death of susceptible hosts. Currently, there is no direct experimental or clinical evidence to support these theories. However, we recently made several potential breakthrough discoveries that substantiate these models. The first is that antibodies against the autolysin (Atl) proteins efficiently induce megacluster formation, opsonophagocytosis, and protection/survival following S. aureus osteomyelitis in mice and humans. The second is that iron sensing determinant B (IsdB) is the most immuno-dominant S. aureus antigen in mouse and man, and that antibodies against IsdB are associated with sepsis, multiorgan failure and death following surgical site infection.8,13 Here we propose to: 1) elucidate the molecular mechanism of anti-IsdB mediated septic death, 2) validate the “susceptible” anti-Isd vs. “protective” anti-Atl immune proteome in PJI patients, and 3) extend this osteoimmunology to elucidate the immune proteomes of S. epidermidis and S. lugdunensis. Our approaches to achieve these goals are embodied by three Specific Aims. In Aim 1 we will elucidate the mechanism of anti-IsdB and related anti-IsdA and anti-IsdH antibody mediated sepsis following S. aureus osteomyelitis. We will also complete a clinical pilot study to demonstrate that osteomyelitis patients who succumb to S. aureus septic death have high anti-Isd titers and Trojan horse macrophages in their blood and internal organs. In Aim 2 we will test the immune proteome hypothesis by correlating anti-Isd and anti-Atl titers with the clinical outcomes of 300 PJI patients that undergo 2- stage revision surgery using our multiplex-Luminex assay, and define the susceptible:protective index of these immune proteomes. And in Aim 3 we will extend our knowledge of the mammalian host response to Staphylococcus species by generate multiplex-Luminex assays for S. epidermidis and S. lugdunensis, and uses them to screen sera from mice and PJI patients as we have done for S. aureus. !

抽象的 人工关节感染 (PJI) 是选择性全关节置换 (TJR) 手术的祸根 其中绝大多数是由葡萄球菌引起的。已知 PJI 的发病率为 1-5%。 这是一个非随机事件，很大程度上由患者的具体因素决定，此外，约 13% 的患者。 感染金黄色葡萄球菌的人会出现败血症并死于多器官衰竭，而其他人则几乎不需要什么就能康复。 除了已确定的宿主易感因素外，还有两种主要理论可以进行干预。 第一个是“免疫蛋白质组”假说，该假说假设一系列特定的蛋白质。 宿主针对金黄色葡萄球菌产生的抗体决定了其对感染的易感性与保护性。 第二个是金黄色葡萄球菌传播的“特洛伊木马”模型，其中白细胞在 手术部位，并将细胞内细菌输送到血流和内脏器官，最终 目前，没有直接的实验或临床证据表明易感宿主败血症死亡。 然而，我们最近取得了一些潜在的突破性发现： 首先证实了这些模型是针对自溶素（ATL）的抗体有效蛋白质。 诱导金黄色葡萄球菌巨簇形成、调理吞噬作用和保护/存活 小鼠和人类的骨髓炎第二个是铁感应决定簇B（IsdB）最多。 小鼠和人中的免疫显性金黄色葡萄球菌抗原，并且针对 IsdB 的抗体是 与败血症、多器官衰竭和手术部位感染后死亡有关。8,13 在这里，我们 提议：1）阐明抗 IsdB 介导的脓毒症死亡的分子机制，2）验证 PJI 患者中“易感”抗 Isd 与“保护性”抗 Atl 免疫蛋白质组，以及 3) 扩展了这一点 骨免疫学阐明表皮葡萄球菌和路邓葡萄球菌的免疫蛋白质组。 实现这些目标的方法体现在三个具体目标中，我们将在目标 1 中阐明。 抗 IsdB 和相关抗 IsdA 和抗 IsdH 抗体介导的脓毒症机制。 我们还将完成一项临床试点研究，以证明骨髓炎。 死于金黄色葡萄球菌败血性死亡的患者具有高抗 ISD 滴度和特洛伊木马 在目标 2 中，我们将测试免疫蛋白质组假设。 通过将抗 Isd 和抗 Atl 滴度与 300 名接受 2- 治疗的 PJI 患者的临床结果相关联 使用我们的多重 Luminex 检测进行阶段修复手术，并定义脆弱性：保护指数 在目标 3 中，我们将扩展我们对哺乳动物宿主的了解。 通过对表皮葡萄球菌和葡萄球菌进行多重 Luminex 检测，对葡萄球菌作出反应。 Lugdunensis，并使用它们筛选小鼠和 PJI 患者的血清，就像我们对金黄色葡萄球菌所做的那样。 ！