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资助申请中的研究人员提供技术，智力和物理资源。该实验室在执行本建议书中针对该堆芯提出的所有任务和分析方面具有丰富的经验，并且自WHEALS项目成立以来一直为该项目提供这些服务。 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、储存和/或运输存档的和新获得的血液、粪便和灰尘标本。此外，对于项目2，核心C将 进行外周血单核细胞（PBMC）的分离、培养和刺激;通过流式细胞术进行树突细胞和调节性T细胞表型分析;通过实时PCR进行RNA分析;以及在刺激培养的PBMC后进行分泌的细胞因子的Bio-PCR测定。核心C的具体目标是：1）作为项目1、2、3和4中收集的血液和灰尘样本的中央收集、储存和处理实验室;将血浆和灰尘样本运送到灰尘和IgE分析中心（核心E）， 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中收集的处理过的灰尘样品运送到Lukacs博士的实验室用于项目3，2）进行经验证的流式细胞术测定，用于树突状细胞和TcB的表型分析，并对项目2中刺激的PBMC培养上清液中分泌的细胞因子进行经验证的生物测定， 3）作为血浆、DNA、RNA、灰尘、粪便和其他生物样品的生物储存库，用于未来的研究，4）向生物统计学和数据管理核心（核心B）提供从流式细胞术和生物统计学分析中获得的数据与Cores D和E以及其他数据源的数据集成，用于手稿和其他赠款申请的统计分析，5）与其他研究者合作，对免疫学检测结果进行解释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.