Systemic and Local Immune Function after Pediatric Thermal Injury

项目摘要/摘要 鉴于儿童烫伤的流行和发病率，我们必须完全了解 并确定与这种疾病相关的所有知识差距。小儿烧伤的免疫生物学状况不佳 明确的和如果理解的，可以打开通往新的治疗靶点的大门。我之前的研究培训，目前 研究顾问委员会，强大的导师和作为一名儿科烧伤外科医生的实践将允许我 获取数据并帮助缩小这一知识鸿沟。这项建议的总体目标是进一步发展我的 成为一名外科科学家，专注于儿科烫伤的免疫生物学，目标是 使用免疫调节治疗以减少不良后果。使用在其他形式的 我们将检验我们的中心假设，即PD-1途径上调是 与儿童烧伤后的免疫抑制和不良后果有关； 通过使用PD-1途径抑制剂，这种免疫抑制在体外是可逆的。热的 受伤的儿童将接受纵向血液采样以及组织床采样 清创和移植手术。PD-1/PD-L1/L2的白细胞表达将通过Flow进行定量 细胞学。先天免疫功能和获得性免疫功能将通过体外刺激试验和流量进行测量 细胞学。创面床巨噬细胞PD-1/PD-L1/L2的表达将与PRO一起检测。 免疫组织化学和激光捕获技术用于炎症和抗炎表型标志物的研究 用mRNA图谱进行显微解剖。此外，我们将确定是否有免疫抑制作用 使用体外方法，热损伤是可逆的。我们将共同孵化来自于 使用PD-1途径抑制剂、GM-CSF或重组IL-7的热损伤儿童。然后，单元格将被 通过体外刺激试验和流式细胞术评价免疫功能的恢复情况。这项研究将 第一个评估PD-1通路在儿童烫伤中的作用。这是特别的 考虑到免疫检查点抑制剂在其他形式的疾病中的临床意义，如 癌症和败血症。这个职业发展奖将产生更多的初步数据，并为我提供 拥有成功竞争未来儿科热疗领域独立资金所需的工具 受伤。

PROJECT SUMMARY/ABSTRACT Given the prevalence and morbidity of pediatric thermal injury it is imperative that we completely understand and define all knowledge gaps related to this disease. The immunobiology of pediatric burn injury is not well defined and if understood can unlock the doors to novel therapeutic targets. My prior research training, current research advisory committee, strong mentorship and practice as a pediatric burn surgeon will allow me to obtain data and help close this knowledge gap. The overall objective of this proposal is to further develop my career into a surgeon-scientist whose focus is on the immunobiology of pediatric thermal injury with goals of using immunomodulation therapy to decrease adverse outcomes. Using methods validated in other forms of pediatric critical illness we will test our central hypothesis that upregulation of the PD-1 pathway is associated with immune suppression and adverse outcomes following pediatric burn injury; and that this immune suppression is reversible ex vivo through the use of PD-1 pathway inhibitors. Thermally injured children will undergo longitudinal blood sampling as well as tissue bed sampling at the time of debridement and grafting procedures. Leukocyte expression of PD-1/PD-L1/L2 will be quantified by flow cytometry. Innate and adaptive immune function will be measured by ex vivo stimulation assays and flow cytometry. Wound bed macrophage expression of PD-1/PD-L1/L2 will be measured along with pro- inflammatory and anti-inflammatory phenotypic markers using immunohistochemistry and laser capture microdissection with mRNA profiling. Furthermore, we will determine if the immune suppressive effects of thermal injury are reversible using ex vivo methods. We will co-incubate peripheral blood leukocytes from thermally injured children with inhibitors of the PD-1 pathway, GM-CSF or recombinant IL-7. Cells will then be evaluated for restoration of immune function via ex vivo stimulation assays and flow cytometry. This study will be the first to evaluate the role of the PD-1 pathway with respect to pediatric thermal injury. This is of particular importance given the clinical implications of immune checkpoint inhibitors in other forms of disease such as cancer and sepsis. This career development award will generate further preliminary data and also provide me with the tools necessary to successfully compete for future independent funding in areas of pediatric thermal injury.