Immunopathology

病理特征将由同一组病理学家对所有研究进行评估和分级，我们认为这是该计划项目的重要资产。只要有可能，抗体和细胞因子探针小组也将具有可比性。这将通过推广新的发现来促进项目之间的交叉受精，并通过比较不同物种和不同器官之间的移植物来深入了解一个项目的结果的一般意义。我们也有一个注解良好的Cynomolgus组织库，里面有我们以前移植的冰冻和石蜡包埋的组织。 实验(&gt；5000个样本)，可以用来扩展和改进我们的分析。 将使用的技术包括常规组织学、免疫组织化学、免疫荧光、电子显微镜和数字载玻片成像形态计量学。免疫过氧化物酶技术与一组单抗将区分渗透的细胞类型，黏附和细胞因子分子和受体，以及激活标记。免疫荧光将用于定位免疫球蛋白和补体的沉积，如 以及使用数字成像对细胞标志物进行双重/三重染色。C4D染色将用于检测体液排斥反应。细胞凋亡/DNA片段化标记物(TUNEL)将评估细胞损伤。协议在每个单独的项目中都有详细的说明。 核心中的病理学研究对于评估同种异体器官移植(心、肺和 肾脏)以及减少炎症和实质损伤的干预效果。这些研究是 对于确定渗透到移植物的细胞的性质和位置、抗体在排斥反应中的作用至关重要 以及治疗的全身性毒性。病理结果将与功能和分子相关。 在项目和其他岩心中进行的研究。此外，将对所有动物进行彻底的尸检， 所有主要器官的样本均按常规和特殊技术进行分析。 我们将比较不同方案下同种异体移植物的炎症、分子和细胞事件。连载 从移植当天开始，将对移植物和淋巴结的样本进行评估，以提供见解。 探讨治疗和诱导耐受的作用机制。免疫病理研究是 设计最少组织样本(2个小块，通常为2x2x2 mm)，用于光学显微镜处理 用福尔马林/石蜡技术和戊二醛/多聚甲醛/EPON进行免疫组织化学 一微米切片和电子显微镜。

The pathology features will be assessed and graded by the same group pathologists for all studies, which we consider an important asset of the Program Project. The panel of antibodies and cytokine probes will also be comparable, whenever possible. This should promote cross-fertilization among the projects by extension of novel findings and provide insights into the general significance of the results in one project by comparing and contrasting grafts in different species and between different organs. We also have a well annotated Cynomolgus tissue bank of frozen and paraffin embedded tissue available from our previous transplantation experiments (>5000 samples), which can be drawn upon to expand and refine our analysis. The techniques that will be utilized include routine histology, immunohistochemistry, immunofluorescence, electron microscopy and digital slide imaging morphometry. Immunoperoxidase techniques with a panel of mAbs will distinguish the infiltrating cell types, adhesion and cytokine molecules and receptors, and activation markers. Immunofluorescence will be used to localize the deposition of immunoglobulin and complement, as well as double/triple staining for cell markers using digital imaging. C4d stains will be used to detect humoral rejection. Markers of apoptosis/DNA fragmentation (TUNEL) will assess cell injury. The protocols are given in detail in each of the individual Projects. The pathology studies in the Core are essential to evaluate the status of the organ allografts (heart, lung and kidney) and the effects of the intervention to reduce inflammation and parenchymal injury. The studies are critical to determine the nature and location of the cells infiltrating the graft, the role of antibodies in rejection and systemic toxicity of treatment. The pathology results will be correlated with functional and molecular studies done in the projects and other cores. In addition, complete necropsies will be done on all animals, with samples from all major organs analyzed by routine and special techniques as indicated. We will compare the inflammatory molecular and cellular events in allografts in the different protocols. Serial samples of the graft and lymph nodes, starting on the day of transplant, will be assessed to provide insights into mechanisms of action of the treatment and tolerance induction. The immunopathologic studies are designed with a minimum of tissue sample (2 small blocks, typically 2x2x2 mm) processed for light microscopy and immunohistochemistry with formalin/paraffin techniques and in glutaraldehyde/paraformaldehyde/epon for one-micron sections and electron microscopy.