Pathology

核心B为病理学和免疫病理学研究的机制解释提供通用设施和专业知识。免疫病理学核心B的广泛、长期目标是深入了解用于诱导移植物存活和耐受的治疗操作的作用机制。正如我们的经验一样，这些研究的见解将补充对类似方案患者临床样本的解释。另一个目标是确定和 评估这些治疗可能引起的任何毒性和并发症。病理学分析的主要作用是记录和表征移植物的状态（浸润的性质、急性或慢性排斥反应的存在，包括体液成分）和方案的全身效应（毒性、并发症）。对方案活检进行的机制研究包括检测移植物和淋巴器官中的Treg细胞，测量移植物中的局部炎症以及在同种异体移植物排斥反应中重要的各种细胞类型（T记忆细胞、树突状细胞）的浸润和活化以及抗体/补体效应。这些在三个项目中详细描述，此外，将进行全面尸检，以确定新治疗策略的疗效和不良反应。 将使用的技术包括常规组织学、免疫组织化学、免疫荧光和根据需要提供的专业技术（电子显微镜、激光捕获）。使用一组mAb的免疫过氧化物酶技术将区分浸润细胞类型、粘附和细胞因子分子和受体，以及使用流式细胞仪定量的活化标志物。 整个载玻片的数字成像。免疫荧光将用于定位免疫球蛋白和补体的沉积，以及用双重/三重染色对细胞进行表型分析。C4d染色将用于检测体液排斥反应。我们有一个注释良好的冷冻和石蜡包埋组织的组织库，可从我们以前的移植实验（>6000个样本）中获得，可用于扩展和完善我们的分析。

Core B provides common facilities and expertise for mechanistic interpretation of pathological and immunopathological studies. The broad, long range objective of the Immunopathology Core B is to provide insights into the mechanism of action of the therapeutic manipulations used to induce grafts survival and tolerance. The insights from these studies will complement the interpretation of clinical samples in patients on similar protocols, as has been our experience. The other goal will identify and evaluate any toxicities and complications that may arise from these treatments. The primary role of the pathological analyses is to document and characterize the status of the graft (nature of infiltrate, presence of acute or chronic rejection, including a humoral component) and the systemic effects of the protocols (toxicity, complications). The mechanistic studies to be done on protocol biopsies include detection of Treg cells in grafts and lymphoid organs, measures of local inflammation in the graft and infiltration and activation of various cell types important in allograft rejection (T memory, dendritic cells) and antibody/complement effects. These are described in detail in the three projects, n addition full necropsies will be performed to determine the efficacy and adverse effects of the novel treatment strategies. The techniques that will be utilized include routine histology, immunohistochemistry, immunofluorescence with specialized techniques available as needed (electron microscopy, laser capture). Immunoperoxidase techniques with a panel of mAbs will distinguish the infiltrating cell types, adhesion and cytokine molecules and receptors, and activation markers quantitated using digital imaging of whole slides. Immunofluorescence will be used to localize the deposition of immunoglobulin and complement, as well as phenotyping cells with double/triple staining. C4d stains will be used to detect humoral rejection. We have a well-annotated tissue bank of frozen and paraffin embedded tissue available from our previous transplantation experiments (>6000 samples), which can be drawn upon to expand and refine our analysis.