Histopathology core

历史学核心将在保罗D。Weilstone MDCRC。核心主任是一位经验丰富的肌肉病理学家（Z。Sahenk，MD，Ph.D，Nationwide Children's Hospital的病理学实验室主任）。项目1将需要分析直接取自DMD患者的肌肉组织。在项目1的目标1中，T细胞浸润将被检查为对肌营养不良蛋白的预先存在的免疫应答的表达。此外，还将对DMD mRNA进行测序，以评估剪接改变与抗肌萎缩蛋白抗原表位之间的关系。在项目1的目标2中，将研究皮质类固醇治疗前后的特异性T细胞免疫。项目1的目的3密切关注AAV-微肌营养不良蛋白递送至肌肉后的基因表达和免疫应答。在项目2中，重点转向使用恒河猴的实验范式。这也是本实验室熟悉的领域，因为来自该动物模型的组织在过去几年中一直在本实验室进行处理。在项目1AAV的目的1中，将在中和抗体耗尽后检查微小肌养蛋白基因递送，以研究T细胞免疫对病毒衣壳的作用。在项目2的目标2中，组织切片将用于帮助确定CD 4+和CD 8 + T细胞对外源转基因产物的作用。 在项目2的最终目标3中，需要进行类似的组织研究来了解可能的免疫抑制剂的作用。在人类和非人类灵长类动物组织的研究过程中，组织学切片将采用标准染色和免疫组织化学进行基因表达和T细胞亚群的免疫标记物，以及与本实验室日常研究相似的蛋白质印迹。本提案的成功取决于本实验室的经验和卓越性，原因如下： 1)每个项目的使命将通过单个实验室的专业知识来促进，该实验室具有内置的质量控制措施，以一致的方式处理所有组织。 2)通过将项目1和项目2中的PI与结果测量分开，将提高结果的客观性。 3)通过由相同的工作人员处理所有组织切片，每个项目之间将具有更大的一致性。

The Histopathology Core will perform a vital function in Paul D. Weilstone MDCRC. The director of the core is an experienced muscle pathologist (Z. Sahenk, MD, Ph.D, Director of the Histopathology Laboratory at Nationwide Children's Hospital). Project 1 will require analysis of muscle tissue taken directly from DMD patients. In Aim 1 of Project 1, T cell infiltration will be examined as an expression of a pre-existing immune response to dystrophin. In addition, sequencing of the DMD mRNA will be performed, in order to assess the relationship between altered splicing and antigenic dystrophin epitopes. In Aim 2 of Project 1, specific T cell immunity will be studied before and after corticosteroid treatment. Aim 3 of Project 1 closely scrutinizes gene expression and immune response following AAV-micro-dystrophin delivery to muscle. In Project 2 emphasis switches to an experimental paradigm using the rhesus macaque. This too is familiar territory to this laboratory because tissue from this animal model has been processed in this laboratory for the past several years. In Aim 1 of project 1 AAV.micro-dystrophin gene delivery will be examined after depleting neutralizing antibodies to study the role of T cell immunity to viral capsid. In Aim 2 of project 2 tissue sections will be used to help define the role of CD4+ and CD8+ T cells against a foreign transgene product. In the final Aim 3 of Project 2 similar tissue studies will be necessary to understand the role of possible immunosuppressant agents. In the course of study of both human and non-human primate tissue, histologic sections will employ standard stains and immunohistochemistry for gene expression and Immunological markers for T cell subsets, and western blots similar to every day studies in this laboratory. The success of this proposal is dependent on the experience and excellence ofthis laboratory for the following reasons: 1) The mission of each Project will be facilitated by the expertise of a single laboratory with built in quality control measures to process all tissues in a consistent manner. 2) The objectivity of the results will be enhanced by separating the Pi's in Projects 1 and 2 from the outcome measures. 3) There will be greater consistency between each of the Projects by having all tissue sections processed by the same staff.