Histopathology core

组织病理学核心将在Paul D. Weilstone MDCRC中发挥重要作用。中心主任是一位经验丰富的肌肉病理学家（Z. Sahenk，医学博士，全国儿童医院组织病理学实验室主任）。项目1将需要分析直接取自DMD患者的肌肉组织。在项目1的目的1中，T细胞浸润将被检查为对肌营养不良蛋白预先存在的免疫反应的表达。此外，将对DMD mRNA进行测序，以评估剪接改变与抗原性肌营养不良蛋白表位之间的关系。在Project 1的Aim 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反应。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这个动物模型的组织在过去几年里一直在本实验室处理。在AAV项目1的目标1中。将在耗尽中和抗体后检测微肌营养不良蛋白基因的传递，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定义CD4+和CD8+ T细胞对抗外来转基因产物的作用。

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.