Tissue Analysis

组织分析

基本信息

批准号：
8326441
负责人：
Timothy W Schacker
金额：
$ 39.59万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-08 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8326441
关键词：
Acquired Immunodeficiency Syndrome Address Adrenal Glands Anatomy Antibodies Anus Architecture Biological Assay Blood Brain CCL19 gene CCL20 gene CD4 Positive T Lymphocytes CXCL10 gene Cell Size Cells Cellular Structures Collaborations Collection DNA Data Data Analyses Flow Cytometry Fluorescent Antibody Technique Frequencies Goals Grant HIV HIV Infections Hematoxylin and Eosin Staining Method Histocytochemistry Histologic Histology Image Analysis Immunohistochemistry In Situ In Situ Hybridization Infection Instruction Interferon Type I Interleukin-1 Interleukin-10 Interleukin-15 Interleukin-7 Kidney Label Laboratories Lead Letters Ligands Liver Location Lung Macrophage Colony-Stimulating Factor Measurement Measures Methods Minnesota Molecular Molecular Analysis Oral cavity Organ Pathologic Pathology Phenotype Population Process Protocols documentation RNA Relative (related person)Research Research Personnel Research Support SIV Sampling Site Specific qualifier value Staining method Stains Structure Surveys T-Lymphocyte Techniques Technology Time Tissues Trichrome stain Tryptophan 2,3 Dioxygenase Universities antiretroviral therapy chemokine collaboratory human tissue innovation latent infection latent persistent infection lymph nodes macrophage performance site viral DNA viral RNA

项目摘要

PROJECT SUMMARY (See instructions): The Tissue Analysis Core of this U19 proposal will provide established histologic and molecular analyses of tissues obtained in each of the projects. Tissues will be processed at the time of collection with samples partitioned and stored according to standard protocols that will be developed and supplied by this Core. Drs. Schacker and Estes will develop specialized staining techniques (i.e., double or triple labels using either chromagenic or fluorescent antibodies) as needed over the duration of the grant period. They have appropriate expertise and laboratory support to accomplish these goals. The Core will provide quantitative analysis of specified features of tissues obtained by each of the projects. This will include quantitative analysis of the size of specific cell populations (identified by chromagenic or fluorescent antibody staining) in a specified anatomic location (e.g., the frequency of macrophages in the parafollicular T cell zone). Quantitative data from these analyses will be combined with other analyses to identify location and frequency of specific cell phenotypes not readily identified by immuno-histochemistry (e.g., combining the size of the total CD4+ T cell population in lymph node with flow cytometry data that measures the relative size of the naive subset of CD4+ T cells can provide an estimate of the absolute size of the naive CD4+ T cell in that tissue). The Core will provide specialized techniques for identifying the location and phenotype of cells that are either vDNA+ or vRNA+, including in situ hybridization (vRNA+ cells) and in situ PCR (vDNA+ cells). Combining either of these two in situ techniques with immunohistochemistry allows identification of the exact phenotype of cells that are infected. Whole organ analysis to identify locations of vRNA+ cells will be done in brain, lung, kidney, liver, adrenals, organs of the GU tract, and the entire gut (mouth to anus) to assist in identification of reservoirs of persistent replication. Molecular analyses of all tissues will be performed to measure the frequency of vDNA+ cells and the frequency of 2-LTR circles and integrated DNA. Collectively these data will be used to estimate 1) sites of persistent replication while on suppressive ARV and 2) size and location of reservoirs of cells harboring latent infection.

项目摘要（请参阅说明）：该U19提案的组织分析核心将提供对每个项目中获得的组织的既定组织学和分子分析。组织将在收集时处理，并根据该核心将开发和提供的标准方案分区和存储样品。博士。 Schacker和Estes将根据需要在赠款期间根据需要开发专门的染色技术（即，使用染色体或荧光抗体使用染色体或荧光抗体）。他们拥有适当的专业知识和实验室支持，以实现这些目标。核心将对每个项目获得的组织的指定特征进行定量分析。这将包括定量在指定的解剖位置（例如，在par骨T细胞区域中巨噬细胞的频率）分析特定细胞群体的大小（通过染色体或荧光抗体染色鉴定）。来自这些分析的定量数据将与其他分析相结合，以识别未通过免疫 - 归化学轻易识别的特定细胞表型的位置和频率（例如，将淋巴结中的总CD4+ T细胞种群与流式细胞仪数据结合在一起，与流式细胞仪数据相结合，可测量CD4+ T细胞的相对大小，可用于CD4+ T细胞的相对大小。该组织中的细胞）。核心将提供专门的技术，用于识别VDNA+或VRNA+的细胞的位置和表型，包括原位杂交（VRNA+细胞）和原位PCR（VDNA+细胞）。将这两种原位技术与免疫组织化学相结合，允许鉴定感染细胞的确切表型。整个器官分析以鉴定VRNA+细胞的位置，将在脑，肺，肾脏，肝脏，肾上腺，肾上腺，肾上腺，整个肠道（口腔到肛门）中进行，以帮助识别持续复制的储层。将对所有组织进行分子分析，以测量VDNA+细胞的频率以及2-LTR圆圈和整合DNA的频率。这些数据将用于估计1）在抑制性ARV和2）具有潜在感染的细胞的大小和位置的同时持续复制的位点。