Core--Pathology

核心--病理学

• 批准号: 7009886
• 负责人: Robert B Colvin
• 金额: $ 15.61万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7009886
• 关键词: biomedical facility; cytokine; electron microscopy; enzyme linked immunosorbent assay; flow cytometry; heart transplantation; histology; immune tolerance /unresponsiveness; immunocytochemistry; immunofluorescence technique; immunopathology; immunoperoxidase; laser capture microdissection; lung transplantation; molecular pathology; monoclonal antibody; pathology; polymerase chain reaction; tissue /cell culture; tissue mosaicism; transplantation immunology

The purpose of the Pathology Core is to provide common facilities and expertise for the immunopathological studies in all three projects. The primary role of the pathological analyses in these allograft protocols 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). In addition, the immunopathologic and molecular studies will give insights into the mechanisms of graft rejection and acceptance with protocol biopsies The techniques that will be utilized include routine histology, immunohistochemistry, immunofluorescence, electron microscopy, PCR, ELISPOT/cell culture, and flow cytometry. 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. RT-PCR will be used to detect and quantify synthesis of key cytokines. Laser capture microdissection will be used for some of these studies in most of the projects to select the appropriate cells. Markers of apoptosis/DNA fragmentation (TUNEL) will assess cell injury. Culture of graft cells will be used with ELISPOT to correlate with the functional studies in the blood. Chimerism will be detected by flow cytometry techniques. The protocols are given in detail in each of the individual Projects. 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.

病理学核心的目的是为所有三个项目的免疫病理学研究提供共同的设施和专业知识。病理分析在这些同种异体移植方案中的主要作用是记录和表征移植物的状态（浸润性的性质，急性或慢性排斥的存在，包括体液成分）以及方案（毒性，并发症）的全身效应。此外，免疫病理学和分子研究将深入了解移植物排斥和接受方案活检的机制。带有MAB面板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，以及激活标记物。免疫荧光将用于定位免疫球蛋白的沉积和 使用数字成像为细胞标记物的补体以及双/三重染色。 C4D污渍将用于检测体液排斥。 RT-PCR将用于检测和量化关键细胞因子的合成。在大多数项目中，将使用激光捕获的微分辨率用于选择适当的细胞。细胞凋亡/DNA碎片的标记（TUNEL）将评估细胞损伤。移植细胞的培养将与ELISPOT一起使用，以与血液中的功能研究相关。嵌合主义将通过流式细胞仪技术检测到。这些协议在每个项目中详细介绍。病理特征将由同一群体病理学家评估和分级所有研究，我们认为这是该计划项目的重要资产。抗体和细胞因子探针的面板将 也可以尽可能可比。这应该通过扩展新发现的扩展来促进项目之间的交叉施用，并通过比较和对比不同物种和不同器官之间的一个项目中结果的总体意义。