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

摘要 骨髓炎是骨科手术的祸根，其中大多数是由金黄色葡萄球菌引起的。 在此之前，P50研究中心翻译骨感染的骨免疫学 （CoRTOBI），众所周知，这些罕见的感染后，择期手术（约1%），我们无法控制 它们（~40%的再感染率）是由于特定的宿主-病原体相互作用。虽然合并症与 获得性免疫和S.金黄色葡萄球菌菌株对临床结果的影响 有待研究。因此，CoRTOBI的项目2使用定制的多重免疫测定法、全基因组免疫测定法和全基因组免疫测定法测试了这一点。 测序（WGS）、动物模型和297例确诊为S. 金黄色葡萄球菌感染。这些结果正式确立了抗Gmd抗体作为保护性抗体和抗IsdB抗体 作为免疫蛋白质组的易感决定因素，几乎所有的人都有易感免疫 蛋白质组因此，我们现在假设：1）在抗Gmd与抗IsdB抗体分泌B中存在缺陷 细胞、浆母细胞和/或长寿命浆细胞（LLPC），其负责天然易感性 免疫蛋白质组2)S.金黄色葡萄球菌菌株特异性因子有助于易感免疫蛋白质组的形成。 并且，3）多价纳米颗粒mRNA疫苗可以逆转易感免疫蛋白质组。三 在本次CoRTOBI更新中，为项目2提出了具体目标。在目标1中，我们将阐明机制 敏感的抗S抗体通过定量抗Gmd与抗IsdB特异性B细胞的金黄色葡萄球菌免疫蛋白质组生成 血中的浆母细胞和骨髓中的LLPC。金黄色骨 感染与未感染的健康对照。在目标2中，我们将完成297 S的WGS。金黄色葡萄球菌 AO-CPP注册，并确定菌株中与最差临床结局相关的独特基因。和 在目标3中，我们将证明多价mRNA纳米颗粒疫苗克服免疫缺陷的功效。 敏感免疫蛋白质组金黄色葡萄球菌，并保护小鼠免受植入物相关的骨髓炎。在 除了Gmd，我们还证明了抗Amd，Hla，CHIPS和SCIN抗体是显着的， 与骨髓炎患者的感染控制相关。鉴于mRNA纳米颗粒的历史性成功 在COVID-19大流行期间，我们将利用这项技术为五个国家开发候选疫苗， 抗原将对单价疫苗进行剂量反应研究，以确定最小mRNA 在血清中可诱导抗体的浓度> 10 ng/ml。那么单价和多价的功效 疫苗将针对最致命的S.使用我们的AO-CPP注册表中的金黄色葡萄球菌菌株 预防和治疗植入物相关骨髓炎的鼠模型。总体影响：我们将 确定易感免疫蛋白质组的机制，建立候选mRNA疫苗 有潜力克服它，并添加S。将金黄色葡萄球菌菌株变量纳入我们的术前多参数列线图 治疗反应预测，可用于个性化预防和治疗。

Abstract Osteomyelitis is the bane of orthopaedic surgery, of which the majority is caused by Staphylococcus aureus. Prior to this P50 Center of Research Translation on the Osteoimmunology of Bone Infection (CoRTOBI), it was known that these rare infections following elective surgery (~1%), and our inability to control them (~40% reinfection rate), are due to specific host-pathogen interactions. While co-morbidities associated with infection were also known, the roles of adaptive immunity and S. aureus strain on clinical outcome had yet to be studied. Thus, Project 2 of CoRTOBI tested this using custom multiplex immunoassays, whole genome sequencing (WGS), animal models, and the AO-CPP Bone Infection Registry of 297 patients with confirmed S. aureus infection. The results formally established anti-Gmd antibodies as protective and anti-IsdB antibodies as susceptible determinants of the immune proteome, and that virtually all people have a susceptible immune proteome. Thus, we now hypothesize that: 1) there is a defect in anti-Gmd vs. anti-IsdB antibody secreting B cells, plasmablasts and/or long-lived plasma cells (LLPCs) that is responsible for the native susceptible immune proteome. 2) S. aureus strain specific factors contribute to susceptible immune proteome genesis. And, 3) multivalent nanoparticle mRNA vaccines can reverse the susceptible immune proteome. Three Specific Aims are proposed for Project 2 in this CoRTOBI renewal. In Aim 1 we will elucidate the mechanism of susceptible anti-S. aureus immune proteome genesis by quantifying anti-Gmd vs. anti-IsdB specific B cells and plasmablasts in blood, and LLPCs in bone marrow, in experimental mice and patients with S. aureus bone infections versus uninfected healthy controls. In Aim 2 we will complete WGS of the 297 S. aureus strains in the AO-CPP registry, and identify unique genes in the strains associated with the worst clinical outcomes. And in Aim 3 we will demonstrate the efficacy of multivalent mRNA nanoparticle vaccines to overcome the susceptible immune proteome against S. aureus, and protect mice from implant-associated osteomyelitis. In addition to Gmd, we have also demonstrated that anti-Amd, Hla, CHIPS & SCIN antibodies are significantly associated with infection control in osteomyelitis patients. Given the historic success of mRNA nanoparticle vaccines during the COVID-19 pandemic, we will use this technology to develop candidate vaccines for the five antigens. Dose response studies will be performed for monovalent vaccines to determine the minimal mRNA concentration that can induce antibodies >10ng/ml in sera. Then the efficacy of the mono and multivalent vaccines will be tested against the most virulent S. aureus strains in the AO-CPP registry using our prophylactic and therapeutic murine models of implant-associate osteomyelitis. Overall Impact: We will identify the mechanism responsible for the susceptible immune proteome, establish candidate mRNA vaccines with potential to overcome it, and add S. aureus strain variables into our multiparametric nomogram for pre-op treatment response prediction, which could be used for personalized prophylaxis and treatments.