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

摘要 骨髓炎是骨科手术的祸根，其中大部分是由金黄色葡萄球菌引起的。 在此之前，P50研究中心翻译了骨感染的骨免疫学研究 (CORTOBI)，据了解，这些罕见的感染在择期手术后(~1%)，以及我们无法控制 它们(约40%的再感染率)是由于特定的宿主-病原体相互作用造成的。虽然与此相关的共病 获得性免疫和金黄色葡萄球菌菌株对临床结果的作用也已为人所知。 有待研究。因此，CoRTOBI的项目2使用定制的多重免疫分析、全基因组来测试这一点 测序(WGS)、动物模型和297例确诊S。 金黄色葡萄球菌感染。结果正式确立了抗GMD抗体为保护性抗体和抗ISDB抗体 作为免疫蛋白质组的易感决定因素，几乎所有人都有易感免疫 蛋白质组。因此，我们现在假设：1)抗GMD和抗ISDB抗体分泌B)存在缺陷 细胞、浆母细胞和/或长寿浆细胞(LLPC)是导致自然易感疾病的原因 免疫蛋白质组。2)金黄色葡萄球菌特异因子参与了易感免疫蛋白质组的形成。 3)多价纳米颗粒基因疫苗可以逆转易感的免疫蛋白质组。三 在此次CoRTOBI更新中，为项目2提出了具体目标。在目标1中，我们将阐明其机制。 易感的抗S抗体。定量检测抗GMD与抗ISDB特异性B细胞对金黄色葡萄球菌免疫蛋白质组的影响 和血液中的浆母细胞，骨髓中的LLPC，在实验小鼠和金黄色葡萄球菌患者中 感染与未感染的健康对照。在目标2中，我们将完成297个金黄色葡萄球菌菌株的WGS AO-CPP注册，并在与最差临床结果相关的菌株中识别独特的基因。和 在目标3中，我们将展示多价信使核糖核酸纳米颗粒疫苗的有效性，以克服 针对金黄色葡萄球菌的敏感免疫蛋白质组，并保护小鼠免受植入相关骨髓炎的影响。在……里面 除了GMD，我们还证明了抗AMD、HL A、CHIPS和SCIN抗体显著 与骨髓炎患者的感染控制有关。鉴于信使核糖核酸纳米颗粒的历史性成功 疫苗在新冠肺炎大流行期间，我们将利用这项技术来开发五种候选疫苗 抗原。将对单价疫苗进行剂量反应研究，以确定最低mRNA量 能在血清中诱导抗体10 ng/ml的浓度。然后是单价和多价的功效 疫苗将针对AO-CPP登记中最强的金黄色葡萄球菌菌株进行测试，使用我们的 种植体相关性骨髓炎的预防和治疗小鼠模型。总体影响：我们将 确定易感免疫蛋白质组的机制，建立候选的mRNA疫苗 有可能克服它，并将金黄色葡萄球菌菌株变量添加到我们的多参数诺模图中进行手术前 治疗反应预测，可用于个体化预防和治疗。

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.