Immuno-Epidemiology Research Laboratory Core

项目总结(见说明)： 免疫流行病学研究实验室核心(核心C)的目的是为本计划项目拨款申请的研究人员提供技术、智力和物质资源。该实验室在执行本提案中为该核心提出的所有任务和分析方面拥有丰富的经验，并自WHEALS项目开始以来一直为其提供这些服务。 作为中央处理实验室，项目1、2、3和4将依靠核心C来处理、跟踪、存储和/或运送已存档和新获得的血液、粪便和灰尘样本。此外，对于项目2，核心C将 进行外周血单个核细胞(PBMCs)的分离、培养和刺激；用多色流式细胞仪进行树突状细胞和调节性T细胞表型鉴定；用realfime PCR进行RNA分析；以及对培养的PBMCs刺激后分泌的细胞因子进行Bio-Plex分析。核心C的具体目标是：1)作为项目1、2、3和4中收集的血液和粉尘样本的中央收集、储存和处理实验室；将血浆和粉尘样本运往粉尘和免疫球蛋白E分析核心(核心E)，以便 项目2、3和4；将粪便样本运送到项目1、2和4的微生物群落分析核心(核心D)；以及将项目2中收集的经过处理的粉尘样本运送到Lukacs博士的实验室用于项目3、2)在项目2中执行用于树突状细胞和树突状细胞表型的经验证的流式细胞仪分析，并在项目2中对刺激的PBMC培养上清中分泌的细胞因子进行经验证的Bio-Plex分析 其目的是以高效、可靠和一致的方式，3)将SEND/E作为血浆、DNA、RNA、灰尘、粪便和其他生物样本的生物储存库，以便在今后的研究中使用；4)向生物统计和数据管理核心(核心B)提供从流式细胞仪和生物组合分析中获得的数据，以便与D和E核心以及其他数据来源的数据整合，用于手稿的统计分析和额外的赠款申请；以及5)与其他研究人员合作，解释免疫学分析结果并将其整合到出版物中。

PROJECT SUMMARY (See instructions): The purpose of the Immuno-Epidemiology Research Laboratory Core (Core C) is to provide a technical, intellectual and physical resource for the investigators in this Program Project Grant application. This laboratory has substantial experience performing all of the tasks and assays that are proposed for this Core in the present proposal and has been providing these services for the WHEALS project since its inception. As the central processing laboratory, Projects 1, 2, 3 and 4 will rely on Core C to process, track, store and/or ship archived and newly acquired blood, stool and dust specimens. Additionally, for Project 2, Core C will perform the isolation, culture and stimulation of peripheral blood mononuclear cells (PBMCs); dendritic cell and regulatory T cell phenotyping by multicolor flow cytometry; RNA analysis by realfime PCR; and Bio-Plex assay of secreted cytokines following stimulation of cultured PBMCs. The Specific Aims of Core C are: 1) To serve as the central collection, storage and processing laboratory for blood and dust samples collected in Projects 1, 2, 3 and 4; to ship plasma and dust samples to the dust and IgE Analysis Core (Core E) for Projects 2, 3, and 4; to ship stool samples to the Microbial Community Analysis Core (Core D) for Projects 1, 2 and 4; and, to ship processed dust samples collected in Project 2 to Dr. Lukacs' lab for use in Project 3, 2) To perform validated flow cytometric assays for phenotyping of dendritic cells and Tregs, and to perform validated Bio-Plex assays of secreted cytokines in stimulated PBMC culture supernatants for Project 2, in an efficient, reliable and consistent fashion, 3) To sen/e as a biorepository for plasma, DNA, RNA, dust, stool and other biological samples for use in future studies, 4) To provide data obtained from flow cytometry and Bio-Plex analyses to the Biostatistics and Data Management Core (Core B) for integration with data from Cores D and E and other data sources for statistical analyses for manuscripts and additional grant applications, and 5) To collaborate with the other investigators in the interpretation and integration of immunologic assay results into publications.