CORE--IMMUNOPATHOLOGY

核心A的目的是为以下方面提供通用设施和专业知识 本项目所有项目的免疫病理研究。这个 病理分析在这些耐受方案中的主要作用是 记录和描述移植物的状态(有无 排斥反应、嵌合体状态(细胞类型、分布)和 方案的全身性影响(毒性、并发症)。此外, 免疫病理分析将使我们深入了解其发病机制。 移植排斥和接受，以及所有大体的常规活组织检查 动物和患者研究，并为分析 异种移植排斥反应的机制。 将使用的技术包括常规组织学， 免疫化学、免疫荧光、原位杂交、电子 显微镜、聚合酶链式反应、细胞培养和流式细胞术。低温恒温器切片将 用免疫过氧化物酶技术与一组单抗一起染色以 鉴别浸润性细胞类型、黏附和细胞因子分子 受体和激活标记物。免疫荧光将用于 定位免疫球蛋白亚型的沉积、补体和凝血 组件。将使用原位杂交技术来确定 关键细胞因子的合成。细胞损伤将通过标记物进行评估 细胞凋亡/DNA片段化(TUNEL)。将使用移植细胞的培养 与血液中的功能研究相关联。世界上的嵌合体 人类和灵长类动物的研究将通过聚合酶链式反应和流式细胞术进行检测 技巧。 病理特征将由同一组进行评估和分级 所有研究的病理学家，我们认为这是 计划项目。抗体和细胞因子探针小组也将 只要有可能，就可以进行比较。这应该会促进异种受精 在项目中推广新的发现并提供见解 通过比较一个项目的结果的总体意义 以及不同物种和不同器官之间的移植物对比。

The purpose of Core A is to provide common facilities and expertise for the immunopathological studies in all projects of the Program Project. The primary role of the pathological analyses in these tolerance protocols is to document and characterize the status of the graft (presence or absence of rejection, the status of chimerism (cell type, distribution) and the systemic effects of the protocols (toxicity, complications). In addition, the immunopathological analysis will give insights into the mechanisms of graft rejection and acceptance, with protocol biopsies in all of the large animal and patient studies and provide essential data for the analysis of mechanisms of xenograft rejection. The techniques that will be utilized include routine histology, immunochemistry, immunofluorescence, in situ hybridization, electron microscopy, PCR, cell culture, and flow cytometry. Cryostat sections will be stained using immunoperoxidase techniques with a panel of mAbs to distinguish the infiltrating cell types, adhesion and cytokine molecules and receptors, and activation markers. Immunofluorescence will be used to localize the deposition of immunoglobin isotypes, complement and clotting components. In situ hybridization will be used to identify the site of synthesis of key cytokines. Cell injury will be assessed by markers of apoptosis/DNA fragmentation (TUNEL). Culture of graft cells will be used to correlate with the functional studies in the blood. Chimerism in the human and primate studies will be detected by PCR and flow cytometry techniques. 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.