CORE--IMMUNOPATHOLOGY

核心--免疫病理学

• 批准号: 6580448
• 负责人: Robert B Colvin
• 金额: $ 20.59万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-01 至 2003-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6580448
• 关键词: animal tissue; biomedical facility; cell sorting; cryostat; flow cytometry; histopathology; human tissue; immune tolerance /unresponsiveness; immunocytochemistry; immunofluorescence technique; monoclonal antibody; tissue /cell culture; transplant rejection

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.

核心A的目的是提供共同的设施和专门知识， 项目所有项目的免疫病理学研究。的 在这些耐受性方案中，病理学分析的主要作用是 记录和描述移植物的状态（存在或不存在） 排斥反应，嵌合状态（细胞类型，分布）和 方案的全身效应（毒性、并发症）。此外，本发明还提供了一种方法， 免疫病理学分析将深入了解 移植物排斥和接受，在所有大的 动物和患者研究，并为分析提供必要的数据， 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、PCR、细胞培养和流式细胞术。冷冻切片将 使用免疫过氧化物酶技术用一组mAb染色， 区分浸润细胞类型、粘附和细胞因子分子 受体和激活标记。免疫荧光将用于 定位免疫球蛋白同种型、补体和凝血的沉积 件.将使用原位杂交来鉴定 关键细胞因子的合成。将通过以下标志物评估细胞损伤： 凋亡/DNA片段化（TUNEL）。将使用移植细胞培养物 与血液中的功能研究相关联。嵌合体 人类和灵长类动物研究将通过PCR和流式细胞术进行检测 技术. 病理学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组也将被 只要有可能，就可以比较。这将促进异花受精 在项目中通过扩展新的发现，并提供见解 通过比较，分析了一个项目的结果的一般意义 以及不同物种和不同器官之间的对比移植。